枯草蛋白酶309突变体吸附作用降低而水解作用增强的制作方法

专利名称：枯草蛋白酶309突变体吸附作用降低而水解作用增强的制作方法
技术领域：
此发明是有关于可用于多种洗涤组合物的新的酶突变体，以及编码这些酶突变体的基因。
背景技术：
酶是自然存在的蛋白中的最大的一类。一般来说，每一种酶催化(加速反应而本身不消耗)一种不同的化学反应。众所周知，蛋白酶作为酶的一种，具有水解其他蛋白质的能力(水解就是将一个化合物断裂为两个或更多的简单化合物，同时在断裂的化学键的两端分别结合一个水分子中的H或OH基团)。通过将自然存在或蛋白质工程得到的蛋白酶作为洗衣洗涤剂制剂的添加剂，已使这种水解蛋白的能力得到利用。许多衣服上污渍都含蛋白成分，因而具广谱效应的蛋白酶可大大增加去除这些污渍的能力。
然而这些蛋白在天然的细菌环境中的功效水平常常不能在相对非天然的洗涤环境中得到实现。具体地说，蛋白酶的一些特征如热稳定性，pH稳定性，氧化稳定性和底物特异性，在酶的天然环境之外并不能得到最佳的利用。
蛋白酶的氨基酸顺序决定了蛋白酶的特征。蛋白酶氨基酸序列的变化可能改变酶的特征，或甚至可能使酶失活，其程度取决于氨基酸序列变化的位点。天然属性和/或变化的程度。一些方法已经被用来改变野生型蛋白酶的氨基酸序列，试图改进它们的特征，以达到增加蛋白酶在洗涤环境中功效的目的。这些方法包括改变氨基酸序列来增加热稳定性和提高其在完全不同的条件下的氧化稳定性。
虽然已有上述本领域的种种方法，但仍然需要新的蛋白酶的突变体，用于洗涤各种表面。
发明目的此发明的目的之一是提供枯草蛋白酶309的突变体，其同野生型酶相比具更强的水解能力。
此发明的另一目的是提供含有这些枯草蛋白酶309突变体的洗涤组合物。
概述此发明涉及枯草蛋白酶309突变体，其具有修饰的野生型枯草蛋白酶309氨基酸顺序，野生型氨基酸序列包含第一、第二、第三、第四、第五、第六6个环状区域，修饰氨基酸序列在其中一个或多个环状区域的特定部位有与野生型枯草蛋白酶不同的氨基酸(即替换)，因而使得枯草蛋白酶309突变体与野生型酶相比，具有对不溶性底物更低的吸附性和更强的水解作用。此发明亦是有关于编码此类枯草蛋白酶309突变体的DNA序列。同时亦是有关于包含此类枯草蛋白酶用于清洁多种表面的组合物的。
附图简述

图1A和1B描述了枯草蛋白酶BPN′和枯草蛋白酶309的氨基酸序列。上行表示来源于淀粉液化杆菌(B.amyloliquefaciens)的枯草蛋白酶BPN′的氨基酸序列，(SEQ ID NO2)，下行表示来源于粘连杆菌(B.lentus)的枯草蛋白酶309的氨基酸序列(SEQ ID NO1)，在枯草蛋白酶309序列上的“*”号表示与枯草蛋白酶BPN′比较特定氨基酸残基的缺失。
发明详述I.枯草蛋白酶309突变体此发明涉及枯草蛋白酶，具体来说是枯草蛋白酶309，通过将编码酶的核苷酸序列的多点突变修饰酶的氨基酸序列，从而修饰了这些酶。此发明中修饰后的枯草蛋白酶(此后称为“枯草蛋白酶309突变体”)，同野生型枯草蛋白酶相比，对于不溶性底物具有更低的吸附作用和更强的水解作用。此发明亦有关于编码这些枯草蛋白酶309突变体的突变基因。
当提到天然枯草蛋白酶309的氨基酸序列时(含269个氨基酸残基)，枯草蛋白酶BPN′经常被用作标准。“BPN′”序列的使用已经成为鉴定所有枯草蛋白酶中氨基酸残基位置的常规方法。天然枯草蛋白酶BPN′和309的氨基酸序列已在图1A和图1B中列出，图中“*”号表示与天然枯草蛋白酶BPN’相比，枯草蛋白酶309氨基酸序列中特定氨基酸残基的缺失。然而，为了在此处讨论的方便，氨基酸位置的参照系是基于“真实”的枯草蛋白酶309的顺序位置(例如，参照SEQ ID NO1，下面加以讨论)此发明的枯草蛋白酶309属于蛋白酶类。蛋白酶是断裂肽键的一种催化剂。有一类蛋白酶称作丝氨酸蛋白酶，此类酶区别特征在于这样一个事实，即在活化区域必须存在一个丝氨酸残基。
酶对于可溶性底物的水解效率随着酶浓度增加而增加这一现象已经得到很好的证实。因而在诸如洗涤过程中遇到的结合于物质表面的底物，水解效率亦可能随着表面浓度的增加而增加，这已得到证明。(BrodeP.F.III和D.S.Rauch，LANGMWR.“枯草蛋白酶BPN″在固定的底物上的活性”8卷，1325-1329页(1992))事实上，当物质表面酶的浓度发生变化时，对于不溶性底物的酶解效率存在着依赖于表面浓度的线性关系(Rubingh，D.N.和M.D.Bauer，“蛋白酶在蛋白溶液分界面上对于水解的催化作用”在多聚物溶液，混合物和分界面。《Elsevier》I.Noda和D.N.Rubingh编，464页，1992)。奇怪的是，当试图运用这一原理来寻找具有更佳功效的突变体蛋白酶时，我们并没有发现吸附更多的酶具有更佳的功效。事实上，我们发现一个相反的事实酶对于底物的吸附作用降低导致底物的水解作用增强(即更佳的洗涤效果)。
由于不愿受到理论的束缚，有理由认为，当比较一种突变体与另一种的功效时，功效的增加是由于吸附至表面越少的酶其结合越不牢固，因而在要去除不溶性蛋白底物的表面具有更高的活动性。在酶溶液浓度相对可比的情况下，这一增加的活动性足以超过任何将更高浓度酶转移至表面所具有的优点。
此处所叙述的突变是设计用来改变(即降低)酶对于表面污渍的吸附作用。在枯草蛋白酶309中，一定位点的氨基酸形成了酶外部的环状区，为便于讨论，这些环状区分别被称作第一、第二、第三、第四、第五环状区。明确地说，57-64位点第一环状区；93-105形成第二环状区；124-131位点形成第三环状区；152-161位点形成第四环状区；181-185位点形成第五环状区；193-214形成第六环个状区域(位点顺序与要枯草蛋白酶309的氨基酸顺序类似(SEQ ID NO1)。
据信这些环状区域在酶分子对于表面结合的肽类物质的吸附上起着重要作用，如果一个或多个环状区域发生突变，对于酶的吸附能力将产生重要影响。由于不愿受到理论的束缚，可以认为这些环状区域对于枯草蛋白酶309分子的吸附作用非常重要，主要在于至少两个原因第一，组成环状区域的氨基酸能紧密地结合于酶分子所接触的表面；第二，这些环状区域处于活性部位以及枯草蛋白酶309的结合口袋区的附近使得他们在酶对于表面结合底物(肽/蛋白质污渍)的催化性吸附作用起到作用。
下表列出了所用到的描述氨基酸的缩写氨基酸三字母缩写符 单字母缩写符丙氨酸AlaA精氨酸ArgR天冬酰胺 AsnN天冬氨酸 AspD天冬酰胺或AspD天冬氨酸半胱氨酸 CysC谷氨酰胺 GlnQ谷氨酸GluE谷氨酰胺或GlxZ谷氨酸甘氨酸GlyG组氨酸HisH异亮氨酸 IleI亮氨酸LeuL赖氨酸LysK甲硫氨酸 MetM苯丙氨酸 PheF脯氨酸ProP丝氨酸SerS苏氨酸ThrT色氨酸TrpW酪氨酸TyrY缬氨酸ValV该位点无氨基酸Xaa*
此文中，“突变体”表示一种与野生型氨基酸序列不同的酶；此文中，“枯草蛋白酶309 DNA突变序列”表示编码一种枯草蛋白酶309突变体的DNA序列；此文中，“野生型枯草蛋白酶309”指由SEQ ID NO1代表的一种酶。枯草蛋白酶309的氨基酸序列在世界专利公开89/06279(1989)中有详细描述，此处以文献形式引用，亦可参看，世界专利公开94/02618，Mullewr等，1994年2月3日出版。
此文中，“枯草蛋白酶309野生型氨基酸序列”包括SEQ ID NO1以及在57-64，93-105，124-131，152-161，181-185，193-214这些位点之外，氨基酸序列有所修饰的SEQ ID NO1序列。
此文中，“更亲水性氨基酸”指其它任何亲水性比下表中所列某个氨基酸亲水性更强的氨基酸。下面这张亲水性表(表1)以亲水性增加的顺序列出了氨基酸。(参看Hopp.T.P和Wods，K.R.，“以氨基酸顺序预测蛋白质的抗原决定簇”《美国科学院院报》78卷，3824-3828页，1981，此处以文献引用)。
表1氨基酸 亲水值Trp -3.4Phe -2.5Tyr -2.3Leu，Ile -1.8Val -1.5Met -1.3Cys -1.0Ala，His -0.5Thr -0.4Pro，Gly -0.0Gln，Asn 0.2Ser 0.3Arg+，Lys+，Glu-，Asp-3.0
表1亦指出哪个氨基酸带电荷(pH8-9条件下的特征)Arg和Lys带正电，Glu和Asp带负电，其余的为中性。在此发明优选用来替代的氨基酸是中性或带负电氨基酸，更优选地带负电(即Glu或Asp)。
因而，举例来说，“用相同或更亲水性氨基酸替换Gln”这句话表示Gln将被Asn(同样亲水性)或Ser、Glu、Asp(具有比Gln更强的亲水性)所替换。每一种氨基酸是中性，或带负电，且具有比Gln更强的亲水性。同理，“用中性或带负电的亲水性更强的氨基酸替换Pro”则表示Pro将被Gln、Asn、Ser、Glu、Asp所替换。
A.第六环状区替换突变体1.包括至少一个氨基酸替换的突变体作为本发明的一个实施方案，枯草蛋白酶309突变体具有修饰的枯草蛋白酶309野生氨基酸序列，该氨基酸序列包括在如下一个或多个位点的替换193，194，195，196，197，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213或214。因而使得枯草蛋白酶309突变体与野生型枯草蛋白酶309相比对于不溶性底物具有更低的吸附性和更强的水解作用。优选地替换的氨基酸位点为，193，194，195，196，199，201，202，203，204，205，206或209；更优选194，195，196，199或201。
优选地，用于替换193位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Pro、Ser、Thr。
优选地用于替换194位的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
优选地，用于替换195位的氨基酸为Asn、Asp、Gln、Glu、Gly、Ser。
优选地，用于替换196位的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
优选地，用于替换197位的氨基酸为Ala、Asp、Cys、Gln、Glu、Gly、His、Met、Pro或Ser。
优选地，用于替换199位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
优选地，用于替换200位的氨基酸为Asn、Asp、Gln、Glu或Ser。
优选地，用于替换201位的氨基酸为Asp或Glu。
优选地，用于替换202位的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
优选地，用于替换203位的氨基酸为Ala、Asn、Asp、Gln、Glu、His、Ile、Met、Pro或Ser。
优选地，用于替换204位的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。
优选地，用于替换205位的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
优选地，用于替换206位的氨基酸为Asp或Gln。
优选地，用于替换207位的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
优选地，用于替换208位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro或Val。
优选地，用于替换209位的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
优选地，用于替换210位的氨基酸为Asp或Glu。
优选地，用于替换211位的氨基酸为Ala、Asn、Asp、Cys、Gln、Gly、His、Ile、Met、Pro、Ser、Thr或Val。
优选地，用于替换212位的氨基酸为Glu。
优选地，用于替换213位的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
优选地，用于替换214位的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
参考表1，优选用于替换193，194，195，196，197，198，199，201，202，203，204，205，206，207，208，209，210，211，212，213和214位的氨基酸为中性或带负电荷氨基酸，并且与野生型枯草蛋白酶309相关位点氨基酸相比具有相同或更强亲水性，其中更优选具更强亲水性的氨基酸。
同样更优选地用于替换193，194，195，196，197，199，200，201，202，204，205，206，207，208，209，210，212，213和214位的氨基酸为Asp或Glu；而用于替换203和211位点的氨基酸为Asp。
2.包括至少两个氨基酸替换的突变体。
作为此发明的另一个实施方案，枯草蛋白酶309突变体具有修饰的枯草蛋白酶309野生氨基酸序列，该氨基酸序列包括在如下两个或更多位点的替换193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213或214。因而使得枯草蛋白酶309突变体与野生型枯草蛋白酶309相比对于不溶性底物具有更低的吸附性和更强的水解作用。优选地被替换的氨基酸位点为，193，194，195，199，201，202，203，204，205，206或209；更优选地194，195，196，211或213。
优选地，用于替换193位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、G1y、His、Met、Pro、Ser或Thr。
优选地，用于替换194位的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
优选地，用于替换195位的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。
优选地，用于替换196位的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
优选地，用于替换197位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
优选地，用于替换198位的氨基酸为Asp、Gln、Glu、或Ser。
优选地，用于替换199位的氨基酸为A1a、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
优选地，用于替换200位的氨基酸为Asn、Asp、Glu或Ser。
优选地，用于替换201位的氨基酸为Asp或Glu。
优选地，用于替换202位的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
优选地，用于替换203位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser或Thr。
优选地，用于替换204位的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。
优选地，用于替换205位的氨基酸为Asn、AspGln、Glu、Pro或Ser。
优选地，用于替换206位的氨基酸为Asp或Glu。
优选地，用于替换207位的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
优选地，用于替换208位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro或Val。
优选地，用于替换209位的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
优选地，用于替换210位的氨基酸为AspGlu。
优选地，用于替换211位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val。
优选地，用于替换212位的氨基酸为Asp、Gln、Glu或Ser。然而，如果211位被Asn、Glu或Val取代，则212位不用Asp取代。
优选地，用于替换213位的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
优选地，用于替换214位的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
参考表1，更优选地用于替换193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213和214位的氨基酸为中性或带负电荷氨基酸，并且与野生型枯草蛋白酶309相关位点氨基酸相比具有相同或更强亲水性，其中优选具更强亲水性的氨基酸。
同样更优选地用于替换193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213和214位点的氨基酸为Asp或Glu。
3.包括至少三个氨基酸替换的突变体作为本发明的一个实施方案，枯草蛋白酶309突变体具有修饰的枯草蛋白酶309野生氨基酸序列，该氨基酸序列包括在如下三个或更多个位点的替换193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213或214。因而使得枯草蛋白酶309突变体与野生型枯草蛋白酶309相比对于不溶性底物具有更低的吸附性和更强的水解作用。优选地被替换的氨基酸位点为，193，194，195，199，201，202，203，204，205，206或209；更优选地194，195，196，211或213。
优选地，用于替换193位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
优选地，用于替换194的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
优选地，用于替换195位的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。
优选地，用于替换196位的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
优选地，用于替换197位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
优选地，用于替换198位的氨基酸为Asn、Gln、Glu、或Ser。
优选地，用于替换199位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
优选地，用于替换200位的氨基酸为Asn、Asp、Glu或Ser。
优选地，用于替换201位的氨基酸为Asp或Glu。
优选地，用于替换202位的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
优选地，用于替换203位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val。
优选地，用于替换204位的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。
优选地，用于替换205位的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
优选地，用于替换206位的氨基酸为Asp或Glu。
优选地，用于替换207位的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
优选地，用于替换208位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Thr或Val。
优选地，用于替换209位的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
优选地，用于替换210位的氨基酸为Asp或Glu。
优选地，用于替换211位的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr中Val。
优选地，用于替换212位的氨基酸为Asp、Gln、Glu、或Ser。
优选地，用于替换213位的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
优选地，用于替换214位的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
参考表1，优选用于替换193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213和214位的氨基酸为中性或带负电荷氨基酸，并且与野生型枯草蛋白酶309相关位点氨基酸相比具有相同或更强亲水性，其中更优选具更强亲水性的氨基酸。
同样更优选地用于替换193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213和214位的氨基酸为Asp或Glu。
B.多个环状区替换的突变体作为此发明的另一项方案，枯草蛋白酶309突变体具有修饰的枯草蛋白酶309野生氨基酸序列，该氨基酸序列包括在1个或多个第1、第2、第3、第4或第5环状区和一个或多个位点的替换因而使得枯草蛋白酶309突变体与其野生型相比，对于不溶性底物的吸附作用降低，水解作用增强。
作为此发明的又一项方案，枯草蛋白酶309突变体还包括第六环状区的一个或多个位点替换。
在此发明的一个优选方案中，根据表1，用于一个或多个环状区的一个或多个位点替换的氨基酸为中性或负电荷，而且与野生型氨基酸序列中相关位点的氨基酸相比，具有相同或更强亲水性，其中选用更强亲水性的氨基酸。
1.第1环状区的替换当一个替换发生在第1环状区时，替换发生在如下一个或多个位点57，58，59，60，61，63，或64。
当一个替换发生在57位点，用于替换的氨基酸为Asn、Asp、Glu或Ser。
当一个替换发生在58位点，用于替换的氨基酸为Glu。
当一个替换发生在59位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
当一个替换发生在60位点，用于替换的氨基酸为Asp、Gln、Glu或Ser。
当一个替换发生在61位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
当一个替换发生在63位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
当一个替换发生在64位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser2.第2环状区的替换当一个替换发生在第2环状区时，替换发生在如下一个或多个位点93，94，95，96，97，98，99，100，101，102，103，104或105。
当一个替换发生在93位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
当一个替换发生在94位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val。
当一个替换发生在95位点，用于替换的氨基酸为Asn、Asp、GlnGlu、Pro或Ser。
当一个替换发生在96位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
当一个替换发生在97位点，用于替换的氨基酸为Asp或Glu。
当一个替换发生在98位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
当一个替换发生在99位点，用于替换的氨基酸为Asp或Glu。
当一个替换发生在100位点，用于替换的氨基酸为Asn、Asp、GlnGlu、Pro或Ser。
当一个替换发生在101位点，用于替换的氨基酸为Asp或Glu。
当一个替换发生在102位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
当一个替换发生在103位点，用于替换的氨基酸为Asp或Glu。
当一个替换发生在104位点，用于替换的氨基酸为Asp或Glu。
当一个替换发生在105位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Leu、Met、Pro、Ser、Thr或Val。
3.第3环状区的替换当一个替换发生在第3环状区时，替换发生在如下一个或多个位点124，125，126，127，128，129，130或131。
当一个替换发生在124位点，用于替换的氨基酸的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val。
当一个替换发生在125位点，用于替换的氨基酸的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
当一个替换发生在126位点，用于替换的氨基酸的氨基酸为Asp或Glu。
当一个替换发生在127位点，用于替换的氨基酸的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。
当一个替换发生在128位点，用于替换的氨基酸的氨基酸为Asp或Glu。
当一个替换发生在129位点，用于替换的氨基酸的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。
当一个替换发生在130位点，用于替换的氨基酸的氨基酸为Asp或Glu。
当一个替换发生在131位点，用于替换的氨基酸的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
4.第4环状区的替换当一个替换发生在第4环状区，替换发生在如下一个或多个位点，152，153，154，155，156，157，158，159，160或161。
当一个替换发生在152位点，用于替换的氨基酸的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
当一个替换发生在153位点，用于替换的氨基酸的氨基酸为Asp、Gln、Glu或Ser。
当一个替换发生在154位点，用于替换的氨基酸的氨基酸为Asp或Glu。
当一个替换发生在155位点，用于替换的氨基酸的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
当一个替换发生在156位点，用于替换的氨基酸的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
当一个替换发生在157位点，用于替换的氨基酸的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
当一个替换发生在158位点，用于替换的氨基酸的氨基酸为Asp或Glu。
当一个替换发生在159位点，用于替换的氨基酸的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Leu、Met、Pro、Ser、Thr或Val。
当一个替换发生在160位点，用于替换的氨基酸的氨基酸为Asp或Glu。
当一个替换发生在161位点，用于替换的氨基酸的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、I1e、Leu、Met、Pro、Ser、Thr或Val。
5.第五环状区的替换。
当一个替换发生在第五环状区，替换发生在如下一个或多个位点181，182，183，184或185。
当一个替换发生在181位点，用于替换的氨基酸的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
当一个替换发生在182位点，用于替换的氨基酸的氨基酸为Asp或Glu。
当一个替换发生在183位点，用于替换的氨基酸的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、SerThr、Tyr或Val。
当一个替换发生在184位点，用于替换的氨基酸的氨基酸为Asp或Glu。
当一个替换发生在185位点，用于替换的氨基酸的氨基酸为Asn、Asp、Gln、Glu或Ser。
6.第6环状区的替换。
当一个替换发生在第6环状区，连同在前述五个环状区的一个或多个环状区中发生的一个或多个替换、193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213或214。
当一个替换发生在193位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
当一个替换发生在194位点，用于替换的氨基酸为Asn、Asp、Cys、Gln、Glu、Gly、His、Pro、Ser或Thr。
当一个替换发生在195位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。
当一个替换发生在196位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
当一个替换发生在197位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
当一个替换发生在198位点，用于替换的氨基酸为Asp、Gln、Glu或Ser。
当一个替换发生在199位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
当一个替换发生在200位点，用于替换的氨基酸为Asn、Asp、Glu或Ser。
当一个替换发生在201位点，用于替换的氨基酸为Asp或Glu。
当一个替换发生在202位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
当一个替换发生在203位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Ser、Thr或Val。
当一个替换发生在204位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。
当一个替换发生在205位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。
当一个替换发生在206位点，用于替换的氨基酸为Asp或Glu。
当一个替换发生在207位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
当一个替换发生在208位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Ser、Thr或Val。
当一个替换发生在209位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
当一个替换发生在210位点，用于替换的氨基酸为Asp或Glu。
当一个替换发生在211位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val。
当一个替换发生在212位点，用于替换的氨基酸为Asp、Gln、Glu或Ser。
当一个替换发生在213位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。
当一个替换发生在214位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser。
C.在环状区以外其它位点的另外的替换与前述同理除了可以在野生型枯草蛋白酶309的第1、第2、第3、第4、第5和/或第6环状区造成替换以外，替换亦可以在环状区以外的位点造成(此指“另外的替换”)。在此发明的另一个方案中，枯草蛋白酶309突变体具有修饰的枯草蛋白酶309野生氨基酸序列，其中除了在上述一个或多个环状区发生一个或多个替换以外，在74位点有一个替换。这一另外的替换既可单独发生(优选地)，亦可与其他“另外的替换”协同发生。
当枯草蛋白酶309突变体另外的替换单独发生在74位点时优选地用于替换的氨基酸为Asn、Asp、Glu、Gly、His、Lys、Phe或Pro，更优选Asp或His。
当枯草蛋白酶309突变体有不止一个另外的替换，优选地与74位点替换协同致替换的位点为如下一个或多个位点的残基，97，99，101，102，105和121。优选地与的另外的替换组合包括74/97；74/99；74/101；74/102；74/105；74/121；74/97/99；74/97/101；74/97/102；74/99/101；74/99/102；74/101/102；74/102/105；74/102/121；74/105/121；74/97/99//101；74/97/99/102/；74/97/101/102；74/99/101/102；74/101/102/121；74/102/105/121；74/97/99/101/102；74/97/101/102/121；和/或74/97/99/101/102/121。最优选地的另外的替换组合包括74/97；74/102；74/97/102；74/101/102；74/102/105；74/99/101/102；74/97/99/101/102和74/99/102。
在每一个鉴定的氨基酸残基位点，其中包括但不限于74位点上造成的另外的替换包括Asp、His、Glu、Gly、Phe、Lys、Pro、和Asn；97位点替换包括Asp、Thr、Asn、Gln、Gly和Ser；99位点替换包括Gly、Asp、Lys、Leu、Ala、Glu和Ser；101位点替换包括Gln、Thr、Asp、Glu、Tyr、Lys、Gly、Arg和Ser；102位点的替换包括Ser、Tyr、Ile、Leu、Met、Ala、Trp、Asp、Thr、Gly和Val；105位点替换包括Val、Leu、Met、Tyr、Gly、Glu、Phe、Thr、Ser、Ala和Ile；121位点的替换包括Asn、Thr、Ile和Ser。更优选地用于替换每一个位点的氨基酸在表2中列出，虽然表2中列出了更优选地的氨基酸，但应明白在同样的位点上任何氨基酸均可用于替换。正如上述讨论的一样这些替换是除了在上述讨论的一个或多个环状区发生的一个或多个替换之外的替换。
表2氨基酸残基优选用于替换或插入的氨基酸74Asp，His97Asp，Thr，Asn，Gly99Arg，Gly，Asp，Lys，Leu，Ala，Glu101Ala，Gln，Thr，Asp，Glu，Tyr，Lys，Gly，Arg102Ile，Tyr，Ser，Leu，Ala，Thr，Gly105Val，Leu，Tyr，Gly，Phe，Thr，Ser，Ala121Ser，Thr，IleD.酶突变体的制备实施例1突变的309 DNA序列构建一种包含野生型枯草蛋白酶309基因的噬菌体质粒(PJMA602)将2.8kbp长度的质粒PUC119的限制酶PvuII片段(vieira，J.和Messing，J.，“单链质粒DNA制备”，153，酶学方法3-11页(1989))，克隆入质粒pUB110的PvuII位点(杆菌遗传贮存中心，哥伦布OH1E9)。此pUC119-pUB110杂交区质粒命名为pJMA601。将通过聚合酶链式反应从粘连杆菌染色体DNA中扩增得来的309(savinase)基因产物克隆入pJMA601的BamHI位点，得到噬菌体质粒pJMA602(国立工业和海洋微生物保藏中心，粘连杆菌10309)噬菌体质粒pJMA602转化大肠杆菌ung株CJ236，并利用VCSU13辅助噬菌体制造含尿嘧啶的单链DNA模板(Kunkel，TA.，JD.Roberts和R.A.Zakour，“通过非表型选择方法快速有效进行定点突变”酶学方法154卷，367-382页)(1987)，经Yuckerbeg，P.D.，F.witney，J.Gieisselsoder和J.McClay修改后的方法“体外利用含尿嘧啶NDA和噬菌体载体进行定点的基因突变，定点的基因突变—一种特殊的方法，M.J.McPherson编，27-48页，(1991))；上述两种方法在此作为文献引用)。作为Zoller和Smith方法的改进，一种单引物的定点基因突变(Zoller，M.J.和M.Smith，“利用MB来源的载体进行寡聚核苷酸定点基因突变一种在任何片段NDA中产生点突变的有效和普遍的方法。”核酸研究，10卷6487-6500页，(1982)，此外引入作为参考)。被用来制造所有的突变体(以上基本由Yuckenberg 1991年提出)。寡核苷酸由AppliedBiosystem Inc.380B DNA合成仪合成。突变反应产物转化进入大肠杆菌菌株MM294(美国典型培养物保藏中心大肠杆菌33625)所有的突变体通过DNA测序确定，分离的DNA转化进入枯草杆菌表达菌株BG 2036(Yang，M.Y.，E.Ferrari和D.J.，Henner(1984)，“中性枯草杆菌蛋白酶基因的克隆和利用克隆产生体外来源的缺失突变”。细菌学杂志，160卷，15-21页)一种在相应环状区带有移码突变终止信号的修饰突变体pJMA602的被用来产生含尿嘧啶的模板。寡聚核苷酸被设计用来恢复203位的正确读码框并在如下位点编码随机替换57，58，59，60，61，63，64，93，94，95，96，97，98，99，100，101，102，103，104，105，124，125，126，127，128，129，130，13 1，152，153，154，155，156，157，158，159，160，161，181，182，183，184，185，193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213和214(构成这些位点所有三联体密码碱基的相等和/或不同摩尔浓度的四种核苷酸混合物得到)。纠正移码突变终止信号并产生功能性酶的突变通过消化酪蛋白的能力来鉴定。DNA序列测定来鉴定随机替换。
实施例2发酵包含一种感兴趣的枯草蛋白酶突变体的枯草杆菌细胞(BG2036)在一升LB-葡萄糖培养液中生长至对数中期并接种入总体积为10升的Biosat ED发酵罐(B.Brawn生命技术公司，Alletown宾西法尼亚州)。发酵基包括酵母提取物，淀粉、消泡剂、缓冲液和微量矿物质(参看《发酵》操作方法、B.McNeil和L.M.Harvey编，1990)发酵过程中培养基保持在pH7.0，加入氯霉素作为抗生素筛选。细胞在37℃生长过夜至吸光值(A600)为大约60然后收获)。
实施例3纯化发酵培养基通过下列步骤获得纯酶。离心去除枯草杆菌细胞并通过100K cutoff膜去除细颗粒。然后浓集于10K cutoff膜，透析降低离子强度并用0.025M MES缓冲液(2-(N-吗啉代)乙烷磺酸)调节pH至5.5。通过离子交换层析柱或亲和吸附层析柱并以氯化钠或丙二醇递度洗脱来进一步纯化酶(参看Scopes，R.K.蛋白纯化原理与实践，Springer-vorlag，纽约(1984)；此处引入作为参考)。
用pNA分析(DelMar，E.G.C.Largman，J.W.Brodrick和M.C.Geokas，生化年鉴，99卷，316-320页(1979)，此以文献引用)来确定梯度洗脱中每部分活性酶的浓度。这种分析法测定的是当酶水解可溶性的合成底物丁二酰-丙氨酰-丙氨酰-脯氨酰-苯丙氨酰对-硝基苯胺(sAAPF-pNA)时释放对-硝基丙胺的速率。由水解作用产生黄色的速率用分光光度计在410nm处测定，它与活性酶的浓度成比例。另外，280nm吸光值测量用来确定总蛋白浓度。活性酶/总蛋白比表示酶的纯度，并用来鉴定收集为贮液的级分。
为避免贮存过程中酶的自我降解，在层析柱上收集的级分中加入等重量丙二醇。纯化过程完成后，酶的贮液纯度通过SDS-PAGE(十二烷基磺酸钠聚丙烯酰胺凝胶电泳)鉴定，酶的绝对浓度利用胰蛋白酶II-T购自西格玛化学公司(圣路易斯，密苏里)的火鸡卵白(egg white)通过活性位点滴定法来确定。测定的转换因子将表明在酶分子中不同位点产生的哪些变化使得酶的突变体对于可溶性底物pNA，比野生型具有更强的活性。
准备使用时，酶的溶液通过Sephadex-G25(Pharmacia，Piscataway新泽西)体积排阻层析柱去除丙二醇并交换缓冲液。贮液中MES缓冲液以含0.01M CaCl2的0.1M的Tris缓冲液替换(三(羟甲基氨基甲烷)，并以HCl调pH至8.6。所有实验在恒温25℃，pH8.6Tris缓冲液中进行。
E.酶突变体的特征实施例4模型表面制备氨丙基可控有孔玻璃(CPG)购自CPG公司，(Fairfield，新泽西)用来作为支持物共价结合购自Bachem公司(Torrence，加利福尼亚)的sAAPF-pNA底物。反应在二甲基亚砜中进行，EDC(1-乙基-3-[3-(二甲氨)丙基]盐酸碳化二亚胺)用作偶联剂。反应一旦完成(pNA分析监测)，多余的溶剂即被去除，以二甲基亚砜(DMSO)和双蒸水清洗CPGsAAPF-pNA。然后在70℃带有N2清洁剂的烘箱中干燥。此反应体系和固定底物制备参照下叙方法进行Brode，P.F.III，和D.S.Raueh.“枯草蛋白酶BPN′在固体底物上的活性”LANGMWR，8卷，1325-1329页，(1992)，此处引入作为参考。CPG表面每平方微米含62000±7000个pNA分子，该表面据CPG公司所报告能承受50.0m2/g压力而保持不变，这就意味着前述将sAAPF-pNA加到CPG上的方法将不破坏孔状结构平均直径为486)实施例5表面水解作用测定使用CPGsAAPF-pNA，一个酶突变体对CPG结合肽的吸附和水解作用可在一次实验中测定。小部分酶突变体的贮液被加入到一个含有Tris缓冲液和已排气的CPGsAAPF-pNA的烧瓶中。烧瓶在一个扭动振荡器中振摇90分钟，期间振荡器在不同的时间间隔内停顿(例如在吸附水解的开始阶段每2分钟-例如，前20分钟并且在进行到实验结尾时每10分钟)。以使CPGsAAPF-pNA沉降和从溶液中取样。实验步骤和吸附与水解作用的计算都参照上述Broder等人1992年描述的方法进行。
实验过程中所有的酶都进行稳定性监测，它们在整个实验过程中应不表现明显的自动分解。这样酶的吸附才可由测定溶液的消耗来确定。起始的酶突变体浓度和独立时间点浓度之间的差异表明了酶吸附的量。表面水解pNA的量通过测定一小份样品在410nm吸光值来测定。pNA水解的总量为样品中的量加上烧瓶内剩余的量。这一数值通过减去没有酶存在情况下，pH8.6由Tris缓冲液水解的pNA量来校正之。这一基础水解依赖于酶的效率，总水解作用在7-29％范围内波动。
实施例6可溶性底物动力学分析对于可溶性底物sAAPF-pNA的水解速率通过Du-70分光光度计在波长410nm处测定吸光值的增加对时间的函数来测定。对每一个动力学测定，酶浓度应保持不变，当底物浓度在90-700uM sAAPF-pNA范围变化时，酶浓度可选择在6-10纳摩尔。在900秒期间内，每1秒测定一个吸光值，数据转换入LotusTM电子表格处理软件(Lotus发展公司，剑桥、马塞诸塞州)。动力学参数分析根据标准的Lineweaver Burk分析法进行，起始(一般来说为第1分钟)部分数据通过线性回归来获取Vo。Vo和So数据以倒数形式绘图获取KM和kcat。
F.枯草蛋白酶309突变体实施例下面的表3-38例举了此发明中枯草蛋白酶309的突变体，这些突变体对于表面结合底物吸附作用降低而水解作用增强。在描述特定的突变时，第一位是原始的野生型氨基酸，第二位表示位点数字，而第三位是用于替换的氨基酸。
表3第1环状区-单突变的突变体Gln57AsnGln57AspGln57GluGln57SerAsp58GluGly59AsnGly59AspGly59GlnGly59GluGly59ProGly59SerAsn60AspAsn60GlnAsn60GluAsn60SerGly61AsnGly61AspGly61GlnGly61GluGly61ProGly61SerGly63AsnGly63AspGly63GlnGly63GluGly63ProGly63SerThr64AsnThr64AspThr64GlnThr64GluThr64GlyThr64ProThr64Ser表4第1环状区-双突变的突变体Gln57Ser＋Asn60GluAsp58Glu＋Gly61GlnGly59Ser＋Gly63SerAsn60Ser＋Gly61SerGly63Asn＋Thr64AspGly59Asn＋Thr64GluAsn60Glu＋Gly63SerAsn60Gln＋Gly63GlnAsn60Asp＋Gly63SerAsp58Glu＋Gly63GlnGln57Asp＋Thr64GlyGln57Glu＋Gly63GlnAsn60Glu＋Gly63AsnGly61Gln＋Gly63AsnAsp58Glu＋Gly59AsnAsp58Glu＋Thr64ProGly61Asn＋Gly63AsnAsp58Glu＋Gly61ProAsn60Glu＋Gly63GlnAsn60Asp＋Gly61GlnAsp58Glu＋Asn60SerGln57Asn＋Gly59GluAsn60Glu＋Gly61ProGly61Asn＋Thr64GlnAsp58Glu＋Thr64SerAsn60Asp＋Thr64GlnGly59Glu＋Gly63SerGln57Asn＋Gly61SerGly61Ser＋Thr64AsnGln57Asp＋Gly61AsnGly59Glu＋Gly63ProAsn60Gln＋Gly61SerGly61Pro＋Gly63SerGly59Asp＋Gly63AsnGly63Gln＋Thr64AsnGln57Glu＋Gly59GlnGln57Asp＋Gly59ProGln57Glu＋Gly61SerGln57Asp＋Gly63GlnAsp58Glu＋Gly61AsnGly59Asn＋Asn60GluGly61Gln＋Thr64GluGln57Glu＋Gly61GlnAsp58Glu＋Gly63SerGly59Glu＋Gly63AsnGly61Ser＋Thr64AspGly59Ser＋Thr64AspGln57Glu＋Gly63ProGly59Ser＋Thr64GluAsn60Gln＋Gly63ProGln57Asn＋Asn60Asp表5第1环状区-三突变的突变体Gly59Pro＋Gly63Gln＋Thr64GluGln575er＋Gly59Asn＋Gly63ProGly59Pro＋Asn60Asp＋Thr64AsnGln57Glu＋Gly59Asn＋Gly63ProGln57Asp＋Gly63Pro＋Thr64AsnGln57Ser＋Asn60Glu＋Thr64GlyGln57Ser＋Asn60Ser＋Gly63SerGly61Ser＋Gly63Ser＋Thr64GlnGln57Glu＋Gly61Pro＋Thr64GlyAsp58Glu＋Gly63Pro＋Thr64GlyGln57Asn＋Asp58Glu＋Gly61ProAsp58Glu＋Gly59Ser＋Thr64ProGln57Asn＋Gly59Glu＋Gly61SerGly59Glu＋Gly61Gln＋Thr64AsnGln57Asp＋Asn60Gln＋Thr64GlyAsn60Glu＋Gly61Gln＋Thr64GlyGln57Asp＋Gly59Pro＋Gly63ProGln57Asn＋Gly63Asn＋Thr64GlyGln57Ser＋Gly59Asp＋Gly61SerGly61Gln＋Gly63Asn＋Thr64ProGln57Glu＋Gly59Pro＋Thr64SerAsn60Asp＋Gly63Gln＋Thr64GlyAsp58Glu＋Gly63Asn＋Thr64GlnAsp58Glu＋Gly61Gln＋Gly63GlnGln57Asp＋Gly61Pro＋Thr64GlnAsp58Glu＋Gly59Asn＋Gly61SerAsp58Glu＋Gly59Ser＋Asn60SerGln57Asn＋Gly61Pro＋Thr64GlyGly59Glu＋Gly61Pro＋Gly63ProGly59Glu＋Gly61Gln＋Gly63AsnGly59Glu＋Gly61Pro＋Gly63GlnGly59Glu＋Gly61Pro＋Thr64AsnGln57Asn＋Asp58Glu＋Thr64Gln表6第1环状区-四突变的突变体Gly59Gln＋Asn60Asp＋Gly61Pro＋Gly63ProGly59Ser＋Asn60Gln＋Gly63Ser＋Thr64AspGly59Pro＋Asn60Ser＋Gly61Ser＋Gly63GlnGln57Glu＋Gly59Pro＋Asn60Ser＋Gly61ProGln57Glu＋Asn60Gln＋Gly63Asn＋Thr64GlyGln57Ser＋Gly59Ser＋Asn60Asp＋Gly61SerGln57Ser＋Asn60Glu＋Gly61Pro＋Thr64ProAsn60Gln＋Gly61Pro＋Gly63Gln＋Thr64AspAsn60Glu＋Gly61Gln＋Gly63Asn＋Thr64ProGln57Glu＋Gly59Gln＋Gly61Pro＋Thr64GlnGln57Asn＋Asp58Glu＋Gly59Ser＋Asn60GlnAsp58Glu＋Asn60Ser＋Gly63Ser＋Thr64SerGln57Glu＋Gly59Pro＋Gly63Gln＋Thr64SerAsp58Glu＋Gly59Ser＋Asn60Ser＋Gly63SerGly59Asn＋Asn60Asp＋Gly61Ser＋Thr64GlnGln57Glu＋Asp58Glu＋Gly59Gln＋Gly61GlnGln57Glu＋Asp58Glu＋Asn60Ser＋Thr64ProGln57Asp＋Asp58Glu＋Gly61Ser＋Gly63ProGln57Asp＋Asp58Glu＋Asn60Gln＋Gly61GlnGln57Ser＋Asp58Glu＋Gly59Glu＋Asn60GlnAsp58Glu＋Gly59Asp＋Gly6lSer＋Gly63GlnGly59Glu＋Asn60Asp＋Gly61Asn＋Gly63AsnGly59Asp＋Asn60Asp＋Gly61Asn＋Gly63GlnGly59Glu＋Asn60Asp＋Gly61Pro＋Thr64SerAsp58Glu＋Gly59Glu＋Asn60Asp＋Thr64GlyAsp58Glu＋Gly59Glu＋Asn60Asp＋Gly61ProAsp58Glu＋Gly59Glu＋Asn60Glu＋Gly61Asn
表7第2环状区-单突变的突变体Val 93AlaVal 93AsnVal 93AspVal 93CysVal 93GlnVal 93GluVal 93GlyVal 93HisVal 93MetVal 93ProVal 93SerVal 93ThrLeu 94AlaLeu 94AsnLeu 94AspLeu 94CysLeu 94GlnLeu 94GluLeu 94GlyLeu 94HisLeu 94IleLeu 94MetLeu 94ProLeu 94SerLeu 94ThrLeu 94ValGly 95AsnGly 95AspGly 95GlnGly 95GluGly 95ProGly 95SerAla 96AsnAla 96AspAla 96GlnAla 96GluAla 96GlyAla 96HisAla 96ProAla 96SerAla 96ThrSer 97AspSer 97GluGly 98AsnGly 98AspGly 98GlnGly 98GluGly 98ProGly 98SerSer 99AspSer 99GluGly100AsnGly100AspGly100GlnGly100GluGly100ProGly100SerSer101AspSer101GluVal102AlaVal102AsnVal102AspVal102CysVal102GlnVal102GluVal102GlyVal102HisVal102MetVal102ProVal102SerVal102ThrSer103AspSer103GluSer104AspSer104GluIle105AlaIle105AsnIle105AspIle105CysIle105GlnIle105GluIle105GlyIle105HisIle105LeuIle105MetIle105ProIle105SerIle105ThrIle105Val表8第2环状区-双突变的突变体Val 93Gln＋Ser 99GluGly 95Ser＋Gly 98GlnSer101Asp＋Ile105AlaLeu 94Ser＋Gly 95SerLeu 94Pro＋Ser101AspGly 98Gln＋Ser103AspSer 97Asp＋Tyr102GlnTyr102Cys＋Ile105MetVal 93Pro＋Gly 98GlnSer 99Glu＋Gly100ProSer103Asp＋Ile105LeuSer 97Asp＋Gly 98GlnVal 93Ser＋Ser 99AspLeu 94Ser＋Gly 98GluGly 95Glu＋Tyr102MetGly 95Asn＋Ser103GluGly 98Ser＋Tyr102ThrGly 98Gln＋Tyr102CysLeu 94Ile＋Ilel05GlnLeu 94Asp＋Gly100GlnAla 96Gly＋Ser101AspSer101Glu＋Ile105GlnGly 95Gln＋Ile105SerVal 93Gln＋Gly100ProVal 93Met＋Ser104AspGly 98Pro＋Ser101AspVal 93Pro＋Ile105HisGly 95Asp＋Gly100SerTyr102Cys＋Ile105LeuGly100Asn＋Ile105MetGly 98Asn＋Ser101GluSer 99Glu＋Ile105GlnVal 93Thr＋Tyr102ThrGly 98Ser＋Ser104GluGly 95Asn＋Ser104GluVal 93Pro＋Ser103GluGly 95Asn＋Tyr102IleLeu 94Pro＋Gly 98AspLeu 94His＋Gly 98AspVal 93Asn＋Tyr102ThrTyr102Ala＋Ser103AspGly 98Pro＋Ser103GluLeu 94Cys＋Tyr102LeuVal 93Gly＋Gly 98SerGly100Gln＋Tyr102SerVal 93Thr＋Gly100AsnGly 98Asn＋Ile105ProVal 93Asp＋Leu 94AlaLeu 94Gly＋Ser104GluVal 93Met＋Ser101AspVal 93Met＋Ser 97GluAla 96Pro＋Ile105AsnSer 97Glu＋Gly 98ProAla 96Thr＋Ser 99GluVal 93Asn＋Gly100AsnGly 95Gln＋Gly 98ProGly 95Asn＋Ala 96HisVal 93Ser＋Ser103AspGly 98Gln＋Ile105ProVal 93Cys＋Ile105Glu
表9第2环状区-三突变的突变体Val 93Gln＋Leu 94Thr＋Ser 99GluLeu 94Met＋Gly 95Gln＋Ser104AspSer 99Glu＋Tyr102Met＋Ile105ThrVal 93Thr＋Leu 94Thr＋Ile105CysLeu 94Asp＋Ala 96Thr＋Gly 98AsnVal 93Met＋Gly 95Gln＋Ser103AspVal 93Cys＋Ala 96Pro＋Ser 99GluLeu 94Asp＋Ala 96Gly＋Gly100GlnLeu 94His＋Gly 98Gln＋Ser 99GluGly 95Pro＋Ser101Asp＋Tyr102GlnLeu 94Asn＋Gly 95Gln＋Ser101AspLeu 94His＋Gly 98Pro＋Ser104GluLeu 94Thr＋Ala 96Asn＋Gly100ProGly 95Ser＋Gly 98Ser＋Tyr102GluAla 96Thr＋Ser 97Asp＋Tyr102ThrLeu 94Asn＋Gly100Ser＋Ser101GluLeu 94Met＋Gly 98Gln＋Gly100AsnVal 93Pro＋Ala 96Glu＋Ile105AlaVal 93Cys＋Gly 95Glu＋Tyr102LeuLeu 94Cys＋Gly 95Ser＋Ser 97AspGly 95Gln＋Ser101Glu＋Tyr102HisVal 93Gln＋Gly 95Glu＋Ile105GlnVal 93Gly＋Ser 99Asp＋Ile105GlyGly 95Asn＋Gly 98Glu＋Ile105ValSer 97Glu＋Gly 98Pro＋Tyr102ThrVal 93Glu＋Leu 94Ile＋Gly100GlnAla 96Pro＋Ser101Asp＋Ile105ProGly 98Pro＋Ser103Asp＋Ile105ValAla 96Gln＋Ser 99Asp＋Tyr102SerSer 97Asp＋Gly 98Ser＋Ile105AsnVal 93Gln＋Gly 98Asn＋Tyr102LeuLeu 94Gly＋Tyr102Pro＋Ser103GluVal 93Thr＋Gly100Gln＋Ser104GluVal 93Gly＋Gly100Gln＋Ser103GluAla 96Thr＋Gly100Ser＋Ser103GluGly100Asn＋Ser101Glu＋Ile105ThrGly 95Asp＋Tyr102Gly＋Ile105MetVal 93Cys＋Ala 96Thr＋Ile105CysAla 96Gly＋Gly 98Asp＋Ser 99AspGly 95Ser＋Ser 97Asp＋Gly 98AspLeu 94Asn＋Ser103Glu＋Ser104AspTyr102Val＋Ser103Glu＋Ser104GluSer 99Asp＋Gly100Glu＋Ile105MetSer 97Asp＋Gly 98Asp＋Ser 99GluGly 95Asn＋Ser101Glu＋Ser103GluSer101Glu＋Tyr102Asn＋Ser103GluLeu 94Asp＋Ser 99Glu＋Gly100GlnGly100Asp＋Ser101Glu＋Ser104GluLeu 94Asp＋Gly 98Glu＋Ile105CysGly 95Glu＋Ser 97Glu＋Tyr102AsnTyr102Thr＋Ser103Glu＋Ile105GluGly 95Asp＋Ser 99Glu＋Gly100GlnGly 95Glu＋Ser 99Glu＋Tyr102MetVal 93Gln＋Ser 97Asp＋Ser 99GluGly100Glu＋Ser101Asp＋Ser103AspVal 93Asp＋Ser103Glu＋Ser104GluVal 93Asp＋Ser 99Asp＋Gly100AspAla 96Gln＋Gly100Asp＋Ser104AspVal 93Asp＋Ser101Glu＋Ile105AspLeu 94Asp＋Ser 99Glu＋Ser101Glu表10第2环状区-四突变的突变体Leu 94Asn＋Ser 97Glu＋Tyr102Asn＋Ile105MetLeu 94Pro＋Gly 95Glu＋Gly100Gln＋Tyr102AlaVal 93Cys＋Leu 94Met＋Gly 95Pro＋Ala 96GlnGly 95Pro＋Ser 97Glu＋Tyr102Gln＋Ile105LeuGly 95Asp＋Gly100Pro＋Tyr102Val＋Ile105CysAla 96Thr＋Gly100Asp＋Tyr102Ala＋Ile105AsnVal 93Asn＋Leu 94Cys＋Ser101Asp＋Tyr102ValVal 93Asn＋Gly 95Asn＋Ser104Asp＋Ile105ValLeu 94Ser＋Gly 98Pro＋Ser103Glu＋Ile105ValVal 93Asn＋Leu 94His＋Gly 95Asn＋Ile105AsnAla 96Ser＋Gly 98Asn＋Ser 99Glu＋Tyr102GlyVal 93Asn＋Gly 98Asn＋Tyr102Ile＋Ile105AspVal 93Asn＋Tyr102Asn＋Ser103Asp＋Ile105SerAla 96Ser＋Gly100Ser＋Ser101Asp＋Tyr102ProLeu 94Pro＋Ala 96Ser＋Ser101Asp＋Tyr102SerVal 93Met＋Ala 96Thr＋Ser104Asp＋Ile105GlnVal 93Met＋Leu 94Ala＋Gly 95Pro＋Ser104GluVal 93Cys＋Gly 95Pro＋Ala 96Glu＋Tyr102ValGly 95Asn＋Ala 96Asn＋Ser104Glu＋Ile105ProVal 93Asn＋Leu 94Asn＋Gly 98Glu＋Ser 99GluLeu 94His＋Gly 98Glu＋Ser 99Glu＋Tyr102SerGly 95Pro＋Gly 98Glu＋Ser 99Asp＋Ile105GlyVal 93Ala＋Ala 96Ser＋Ser103Glu＋Ser104GluLeu 94Gly＋Ser 97Asp＋Gly 98Asp＋Ser 99GluVal 93Cys＋Leu 94Asp＋Gly 95Asn＋Gly100GluVal 93Ser＋Ser101Glu＋Ser103Asp＋Ser104AspAla 96Ser＋Ser101Glu＋Tyr102Gln＋Ser103AspVal 93Thr＋Gly100Asn＋Ser101Glu＋Ser103AspVal 93Asn＋Leu 94Glu＋Gly 95Glu＋Ser 99GluLeu 94Asp＋Ala 96Gly＋Ser 99Glu＋Ile105CysLeu 94Ala＋Ser101Glu＋Ser104Asp＋Ile105SerAla 96His＋Ser101Glu＋Ser104Glu＋Ile105AlaVal 93Gly＋Gly100Pro＋Ser101Glu＋Ser104AspGly 95Gln＋Ser101Glu＋Ser104Asp＋Ile105MetGly 98Gln＋Ser101Glu＋Ser104Glu＋Ile105AsnVal 93Gly＋Ser101Asp＋Tyr102Glu＋Ile105AspSer101Asp＋Tyr102Val＋Ser104Glu＋Ile105GluVal 93Cys＋Gly 95Asp＋Gly 98Asp＋Gly100ProGly 95Asp＋Ser 97Glu＋Tyr102Ala＋Ile105CysVal 93Thr＋Gly 95Glu＋Ala 96Asn＋Ser 99AspVal 93Cys＋Gly 95Asp＋Ser 99Glu＋Ile105ValVal 93Cys＋Ser101Asp＋Ser103Glu＋lle105AspAla 96His＋Ser 97Glu＋Ser 99Asp＋Ile105AsnAla 96Asn＋Ser 97Glu＋Ser 99Glu＋Tyr102ProVal 93Cys＋Ser 97Glu＋Ser 99Asp＋Ile105HisVal 93Ser＋Ser 97Glu＋Gly 98Gln＋Ser 99GluGly 95Asn＋Ser 97Asp＋Ser 99Glu＋Gly100GluVal 93Met＋Gly100Asp＋Tyr102Asp＋Ser103AspLeu 94Gly＋Ser101Asp＋Tyr102Ala＋Ile105GluSer 97Glu＋Gly 98Asp＋Gly100Asp＋Tyr102GlyLeu 94Ser＋Ser 99Glu＋Gly100Glu＋Ser104GluVal 93Asn＋Ser 99Glu＋Gly100Glu＋Ser104AspVal 93Ala＋Ser 99Glu＋Gly100Asp＋Ser104AspLeu 94Cys＋Gly 95Ser＋Ser 99Asp＋Ser101GluGly 95Ser＋Ala 96Thr＋Ser 99Glu＋Ser101AspLeu 94Cys＋Ala 96Ser＋Ser 99Asp＋Ser101AspSer 99Asp＋Ser101Glu＋Tyr102Asn＋Ile105ValVal 93Glu＋Gly 95Pro＋Ser101Glu＋Ser103AspVal 93Ala＋Gly 95Asp＋Ser 99Asp＋Ser101GluSer 99Glu＋Ser101Glu＋Tyr102Pro＋Ser104Glu表11第3环状区-单突变的突变体Leu124AlaLeu124AsnLeu124AspLeu124CysLeu124GlnLeu124GluLeu124GlyLeu124HisLeu124IleLeu124MetLeu124ProLeu124SerLeu124ThrLeu124ValGly125AsnGly125AspGly125GlnGly125GluGly125ProGly125SerSer126AspSer126GluPro127AsnPro127AspPro127GlnPro127GluPro127GlyPro127SerSer128AspSer128GluPro129AsnPro129AspPro129GlnPro129GluPro129GlyPro129SerSer130AspSer130GluAla131AsnAla131AspAla131GlnAla131GluAla131GlyAla131HisAla131ProAla131SerAla131Thr表12第3环状区-双突变的突变体Leu124Gln＋Ser128GluGly125Ser＋Ser126GluGly125Ser＋Ser128GluPro127Gly＋Ser130GluLeu124Thr＋Ser126GluSer126Glu＋Pro129SerGly125Gln＋Ser128GluPro127Ser＋Ser130GluGly125Glu＋Pro127AsnPro129Gln＋Ser130GluGly125Asp＋Ala131HisPro127Gly＋Ala131GluLeu124His＋Ser126AspPro127Gly＋Pro129AspSer126Glu＋Ala131GlnLeu124Asp＋Ala131ProGly125Pro＋Pro127GluGly125Ser＋Ser126AspLeu124Gln＋Ala131AsnGly125Asp＋Pro129SerPro129Asn＋Ala131AspPro127Asn＋Ser128AspPro129Gly＋Ala131GluPro127Ser＋Pro129AspPro127Ser＋Ser128GluLeu124Gly＋Ser130GluGly125Asn＋Ser130AspLeu124Glu＋Pro127AsnSer130Glu＋Ala131GlySer130Asp＋Ala131ThrLeu124Gln＋Ser128AspLeu124Ser＋Ala131ProPro129Ser＋Ser130GluGly125Gln＋Pro127AspLeu124Cys＋Ala131ThrGly125Gln＋Pro129GlnLeu124Met＋Ala131GlnGly125Gln＋Ala131AspSer128Glu＋Ala131AsnSer130Glu＋Ala131ProGly125Pro＋Ala131SerPro127Ser＋Pro129AsnSer126Glu＋Pro127GlnPro127Ser＋Ser130AspPro127Gly＋Ser128AspGly125Glu＋Pro127SerLeu124Cys＋Pro129AsnGly125Gln＋Pro129GluGly125Asn＋Ala131AspGly125Ser＋Pro129GlnSer126Asp＋Pro129GlnLeu124Val＋Pro129AsnLeu124His＋Ser126GluPro127Glu＋Ala131GlyLeu124Thr＋Gly125ProLeu124His＋Ser130GluGly125Asn＋Ser126AspPro129Asn＋Ala131ProPro127Gln＋Pro129AsnLeu124Gly＋Pro129Glu表13第3环状区-三突变的突变体Gly125Gln＋Ser126Glu＋Ala131HisSer126Asp＋Pro127Ser＋Ala131HisLeu124Val＋Ser130Asp＋Ala131AsnLeu124Gln＋Ser126Glu＋Ala131SerGly125Ser＋Pro129Glu＋Ala131GlyLeu124Ser＋Pro127Asp＋Ala131GlnPro127Ser＋Ser128Asp＋Ala131HisLeu124Thr＋Pro127Glu＋Ala131SerLeu124Gln＋Ser126Glu＋Ala131AsnGly125Asn＋Pro127Glu＋Ala131GlnGly125Gln＋Ser126Asp＋Pro127GlnPro127Asn＋Ser130Asp＋Ala131ThrGly125Asn＋Pro127Glu＋Pro129AsnLeu124Thr＋Ser128Asp＋Ala131GlnGly125Ser＋Pro129Gln＋Ala131GluPro127Gln＋Pro129Asn＋Ser130GluLeu124Ile＋Gly125Gln＋Ser130GluPro129Asn＋Ser130Glu＋Ala131GlyLeu124Ser＋Gly125Ser＋Pro129GlnLeu124Asn＋Gly125Gln＋Ser128AspLeu124Gln＋Pro127Ser＋Ser130AspGly125Ser＋Pro127Ser＋Ala131AspGly125Ser＋Ser128Glu＋Pro129GlnPro127Asn＋Ser128Asp＋Ala131GlyLeu124Thr＋Gly125Ser＋Ala131AsnGly125Gln＋Pro129Ser＋Ser130AspGly125Pro＋Ser126Glu＋Pro129AsnPro129Gly＋Ser130Asp＋Ala131ProGly125Asn＋Ser126Asp＋Pro129GlyGly125Asp＋Ser126Asp＋Ala131AsnGly125Glu＋Ser126Glu＋Ala131SerGly125Glu＋Ser126Asp＋Ala131HisGly125Gln＋Ser128Glu＋Pro129GluSer128Glu＋Pro129Glu＋Ala131GlyLeu124Cys＋Ser128Glu＋Pro129AspPro127Gly＋Ser128Glu＋Pro129GluPro127Gly＋Ser128Asp＋Pro129AspLeu124Asn＋Ser128Glu＋Pro129AspSer126Glu＋Pro127Asp＋Ala131AsnSer126Glu＋Pro127Glu＋Pro129GlyLeu124His＋Ser126Asp＋Pro127GluSer126Asp＋Pro127Glu＋Pro129GlyPro127Asp＋Ser128Asp＋Pro129AsnPro127Asp＋Ser128Glu＋Ala131HisPro127Glu＋Ser128Asp＋Ala131ProLeu124Ile＋Pro127Glu＋Ser128GluPro129Asp＋Ser130Asp＋Ala131GlyLeu124Pro＋Pro129Glu＋Ser130GluSer126Asp＋Pro127Glu＋Ser128AspSer126Glu＋Pro127Asp＋Ser128AspPro129Glu＋Ser130Asp＋Ala131GluLeu124Asp＋Gly125Asp＋Ser126AspLeu124Ser＋Ser126Asp＋Ser128AspSer126Glu＋Ser128Asp＋Pro129AsnGly125Asn＋Ser126Glu＋Ser128GluSer126Glu＋Ser128Glu＋Ala131ThrLeu124Asn＋Ser126Glu＋Ser128GluSer126Glu＋Ser128Glu＋Pro129GluGly125Glu＋Ser126Glu＋Ser128AspSer128Asp＋Pro129Ser＋Ser130Glu表14第3环状区-四突变的突变体Gly125Gln＋Ser126Glu＋Pro127Ser＋Ala131SerGly125Asn＋Pro127Ser＋Ser130Asp＋Ala131AsnLeu124Ala＋Pro127Gln＋Ser128Glu＋Ala131ProGly125Pro＋Pro127Ser＋Pro129Ser＋Ser130GluGly125Asn＋Pro129Asn＋Ser130Asp＋Alal31GlnGly125Asn＋Pro129Asn＋Ser130Glu＋Ala131ProLeu124Ser＋Ser126Glu＋Pro129Gly＋Ala131GlnGly125Asn＋Ser126Glu＋Pro127Gly＋Ala131SerLeu124Ile＋Ser128Asp＋Pro129Gln＋Ala131SerLeu124His＋Gly125Ser＋Pro129Gly＋Ala131AsnLeu124Ala＋Gly125Glu＋Pro127Gln＋Pro129GlnLeu124Ser＋Gly125Glu＋Ser126Asp＋Pro129SerGly125Glu＋Ser126Glu＋Pro127Gln＋Ala131HisLeu124Met＋Gly125Asp＋Ser126Asp＋Pro127AsnLeu124Gln＋Ser128Glu＋Pro129Asp＋Ala131GlnLeu124Val＋Ser128Glu＋Pro129Glu＋Ala131AsnPro127Ser＋Ser128Asp＋Pro129Asp＋Ala131ThrLeu124Ile＋Ser128Glu＋Pro129Asp＋Ala131SerGly125Ser＋Ser126Glu＋Pro127Glu＋Ala131GlnLeu124His＋Gly125Gln＋Ser130Glu＋Ala131GluLeu124Thr＋Gly125Gln＋Ser130Asp＋Ala131AspLeu124His＋Pro127Glu＋Ser128Glu＋Ala131HisGly125Ser＋Pro127Asp＋Ser128Asp＋Pro129GlnLeu124Asn＋Ser126Glu＋Pro127Asp＋Ser128AspPro127Gln＋Ser128Glu＋Pro129Asp＋Ser130AspGly125Pro＋Ser128Asp＋Pro129Glu＋Ser130GluLeu124Asn＋Gly125Asp＋Ser126Asp＋Pro127AspLeu124Asp＋Gly125Asp＋Ser126Asp＋Pro129GlnLeu124Asp＋Gly125Glu＋Ser126Asp＋Pro129GlyGly125Gln＋Ser126Glu＋Ser128Glu＋Ala131ThrLeu124Asn＋Ser126Glu＋Pro127Gln＋Ser128AspLeu124Met＋Gly125Pro＋Ser126Asp＋Ser128GluLeu124Asn＋Gly125Ser＋Ser126Asp＋Ser128AspLeu124Val＋Ser126Glu＋Ser128Glu＋Pro129AspLeu124Cys＋Ser126Asp＋Ser128Glu＋Pro129GluSer126Asp＋Pro127Asp＋Pro129Glu＋Ala131SerGly125Asn＋Ser126Glu＋Pro127Glu＋Pro129GluLeu124Ser＋Ser126Asp＋Pro127Asp＋Pro129AspGly125Asp＋Ser126Glu＋Ser128Asp＋Pro129SerLeu124Val＋Gly125Asp＋Ser126Glu＋Ser128GluGly125Glu＋Ser126Glu＋Ser128Asp＋Pro129AsnGly125Glu＋Ser126Asp＋Pro127Ser＋Ser128AspLeu124Met＋Ser128Glu＋Ser130Glu＋Alal31SerLeu124Cys＋Ser128Glu＋Pro129Ser＋Ser130AspGly125Gln＋Ser128Glu＋Ser130Glu＋Ala131HisLeu124Met＋Pro127Gln＋Ser128Glu＋Ser130GluLeu124Gly＋Ser126Asp＋Pro129Glu＋Ser130AspGly125Asn＋Ser126Glu＋Pro129Glu＋Ser130GluLeu124Gly＋Gly125Asp＋Pro127Asp＋Pro129GlySer126Asp＋Ser128Asp＋Ser130Glu＋Ala131GlnGly125Pro＋Ser126Asp＋Ser128Glu＋Ser130AspSer126Asp＋Pro127Gly＋Ser128Glu＋Ser130GluSer126Glu＋Pro127Gly＋Ser128Asp＋Ser130GluSer126Asp＋Ser128Asp＋Pro129Ser＋Ser130AspSer126Glu＋Ser128Asp＋Ser130Asp＋Ala131HisSer126Glu＋Ser128Glu＋Ser130Glu＋Ala131SerGly125Pro＋Ser126Glu＋Ser128Glu＋Ser130AspLeu124Val＋Ser126Asp＋Ser128Glu＋Ser130AspGly125Gln＋Ser126Glu＋Ser128Asp＋Ser130GluSer126Glu＋Ser128Asp＋Pro129Gly＋Ser130Glu
表15第4环状区-单突变的突变体Gly152AsnGly152AspGly152GlnGly152GluGly152ProGly152SerAsn153AspAsn153GlnAsn153GluAsn153SerSer154AspSer154GluGly155AsnGly155AspGly155GlnGly155GluGly155ProGly155SerAla156AsnAla156AspAla156GlnAla156GluAla156GlyAla156HisAla156ProAla156SerAla156ThrGly157AsnGly157AspGly157GlnGly157GluGly157ProGly157SerSer158AspSer158GluIle159AlaIle159AsnIle159AspIle159CysIle159GlnIle159GluIle159GlyIle159HisIle159LeuIle159MetIle159ProIle159SerIle159ThrIle159ValSer160AspSer160GluTyr161AlaTyr161AsnTyr161AspTyr161CysTyr161GlnTyr161GluTyr161GlyTyr161HisTyr161IleTyr161LeuTyr161MetTyr161ProTyr161SerTyr161ThrTyr161Val表16第4环状区-双突变的突变体Ser154Glu＋Gly155AsnSer154Asp＋Gly157GlnSer154Glu＋Ile159HisGly152Gln＋Gly157AspSer154Glu＋Gly155SerAsn153Ser＋Ser154GluAla156Asn＋Ser160GluAla156Thr＋Ser158GluAsn153Glu＋Gly157SerGly155Asn＋Ala156AspIle159Gly＋Tyr161GlyGly155Ser＋Tyr161GluSer154Asp＋Gly155ProGly155Asp＋Ala156SerGly152Glu＋Asn153SerSer158Glu＋Ile159AsnAsn153Asp＋Ala156HisSer154Glu＋Tyrl61CysIle159Leu＋Tyr161AsnGly157Asp＋Ile159GlySer158Asp＋Ile159GlyGly152Pro＋Gly155GluGly155Gln＋Ser160AspGly152Asp＋Tyr161MetSer154Glu＋Gly155GlnGly155Glu＋Tyrl61AlaAla156Ser＋Ser160AspAla156Glu＋Tyr161LeuGly152Pro＋Gly157AsnSer158Glu＋Ile159SerGly157Asp＋Tyr161ThrSer158Asp＋Ile159CysGly155Pro＋Tyr161AlaGly152Asp＋Tyr161AlaAla156His＋Ile159AsnSer154Asp＋Ala156HisGly157Pro＋Ser158AspAsn153Ser＋Ile159CysSer154Asp＋Ala156SerSer154Glu＋Tyr161AlaIle159Leu＋Tyr161AlaGly152Glu＋Gly155GlnAla156Pro＋Gly157GluAsn153Asp＋Tyr161HisAla156Gln＋Ile159AsnGly152Pro＋Ser154AspGly155Gln＋Ala156AspAla156Ser＋Ile159GlyAsn153Asp＋Gly157GlnIle159Leu＋Ser160AspGly155Glu＋Ile159GlnSer160Asp＋Tyr161ThrSer158Asp＋Ile159GlnSer154Glu＋Gly157AsnAla156Gly＋Ile159HisAla156Ser＋Ser158AspAsn153Gln＋Gly155GluGly155Asn＋Ser158GluSer158Glu＋Ile159ProSer158Asp＋Ile159Leu表17第4环状区-三突变的突变体Ala156Asn＋Gly157Asn＋Tyr161CysGly152Gln＋Gly155Glu＋Tyr161AsnGly152Gln＋Asn153Gln＋Ser154GluGly155Ser＋Ser158Glu＋Tyr161GlnGly152Glu＋Asn153Ser＋Ala156SerGly152Glu＋Ile159Pro＋Tyr161AsnGly152Asp＋Gly157Gln＋Tyr161MetGly155Asn＋Ile159Thr＋Tyr161HisAsn153Ser＋Gly155Ser＋Ser158GluGly152Ser＋Gly155Pro＋Ser158GluGly152Pro＋Gly157Gln＋Ser160AspGly152Gln＋Ser154Asp＋Tyr161MetAla156Pro＋Ile159Asn＋Tyr161IleIle159Asn＋Ser160Glu＋Tyr161SerGly152Asp＋Ile159Ala＋Tyr161CysAsn153Gln＋Ile159Ala＋Ser160GluAsn153Asp＋Gly155Pro＋Tyr161AsnGly155Asn＋Ala156Thr＋Ser158AspGly152Gln＋Ser154Asp＋Tyr161GlnGly157Ser＋Ser158Asp＋Tyr161ProSer154Glu＋Gly155Ser＋Ile159CysGly155Gln＋Ala156Asn＋Ser158AspGly155Asn＋Gly157Asn＋Ile159AlaGly157Asn＋Ser158Glu＋Ile159HisAsn153Ser＋Ile159Leu＋Ser160GluAla156Gln＋Gly157Asp＋Tyr161AsnGly155Asn＋Ala156His＋Tyr161ValAsn153Gln＋Ala156Asp＋Ile159GlyAla156His＋Ser160Glu＋Tyr161HisAla156Gln＋Ser160Asp＋Tyr161AlaGly152Asp＋Gly157Gln＋Ile159GlyAsn153Glu＋Ala156Pro＋Ile159AlaGly152Gln＋Gly155Asn＋Ala156ThrAsn153Gln＋Ile159Met＋Ser160GluGly152Asp＋Ala156Pro＋Tyr161CysGly152Ser＋Ser154Glu＋Tyr161ValGly155Asp＋Gly157Ser＋Tyr161AsnAsn153Glu＋Ser154Asp＋Gly155SerAsn153Glu＋Ser154Asp＋Ala156ProGly157Asn＋Ser160Asp＋Tyr161AspGly152Glu＋Asn153Asp＋Tyr161MetGly152Asp＋Asn153Asp＋Ile159LeuAla156Asp＋Gly157Glu＋Tyr161ThrAsn153Ser＋Ser154Glu＋Gly155AspSer154Asp＋Gly155Asp＋Ala156ProAsn153Ser＋Ser154Asp＋Gly155GluSer154Glu＋Gly155Asp＋Tyr161AlaGly157Asp＋Ser158Glu＋Tyr161MetGly155Gln＋Gly157Asp＋Ser158GluAla156His＋Gly157Glu＋Ser158AspGly155Asp＋Ala156Glu＋Gly157AspSer154Asp＋Ala156Glu＋Tyr161CysSer154Asp＋Ala156Glu＋Ile159ProSer154Asp＋Ala156Glu＋Ile159AsnGly152Pro＋Ala156Glu＋Ser158GluGly155Asn＋Ala156Glu＋Ser158GluAsn153Glu＋Gly155Asp＋Ile159ProGly152Glu＋Ile159Cys＋Ser160AspGly152Glu＋Ser154Glu＋Gly157GlnGly152Glu＋Ser154Glu＋Gly155Asn表18第4环状区-四突变的突变体Ala156Pro＋Ile159Pro＋Ser160Glu＋Tyr161ThrAsn153Gln＋Gly157Asn＋Ile159Gln＋Ser160AspAsn153Gln＋Ala156Gly＋Gly157Glu＋Ile159AsnGly152Asn＋Ile157Gln＋Ser160Asp＋Tyr161MetGly152Pro＋Ser157Glu＋Gly157Pro＋Ile159AlaAla156Ser＋Gly157Ser＋Ile159Gln＋Tyr161GluAsn153Ser＋Gly157Ser＋Ile159Ala＋Ser160AspGly155Gln＋Ser158Asp＋Ile159Gly＋Tyr161MetGly152Ser＋Ser154Asp＋Ala156Gln＋Ile159CysAla156His＋Gly157Pro＋Ser158Glu＋Tyr161MetAla156Pro＋Gly157Asn＋Ile159Met＋Ser160AspSer154Glu＋Gly155Asn＋Ile159Met＋Tyr161AlaGly152Ser＋Ala156His＋Gly157Pro＋Ile159GlyAsn153Ser＋Ile159Leu＋Ser160Glu＋Tyr161ThrGly152Asn＋Ser154Asp＋Ala156Ser＋Gly157AsnSer154Asp＋Gly155Gln＋Ile159Leu＋Tyr161ThrAsn153Ser＋Ser154Asp＋Gly155Asn＋Ile159MetGly152Asn＋Asn153Gln＋Gly155Asn＋Ile159ValAsn153Glu＋Ser154Glu＋Gly157Asn＋Ile159LeuAsn153Glu＋Ser154Asp＋Gly155Pro＋Ala156SerSer154Asp＋Gly155Asp＋Ala156Gly＋Gly157AsnGly152Ser＋Gly157Asp＋Ser158Asp＋Ile159SerGly155Pro＋Gly157Asp＋Ser158Asp＋Ile159ProAla156Glu＋Gly157Glu＋Ser158Glu＋Tyr161IleAsn153Glu＋Ser154Asp＋Gly155Asp＋Ala156ProSer154Glu＋Ala156Glu＋Gly157Ser＋Ile159SerAla156Glu＋Gly157Asn＋Ser158Asp＋Ile159AsnGly152Asn＋Asn153Asp＋Gly155Glu＋Tyr161GlnGly152Pro＋Asn153Asp＋Gly155Glu＋Gly157GlnGly152Asp＋Ala156Pro＋Ile159Val＋Ser160AspGly152Asp＋Asn153Gln＋Ile159Ala＋Ser160AspGly152Glu＋Ser154Glu＋Ile159Gln＋Tyr161CysGly152Glu＋Ser154Asp＋Gly157Pro＋Tyr161LeuGly155Asp＋Ala156Asp＋Gly157Pro＋Ser158AspGly155Glu＋Ala156Asp＋Gly157Gln＋Ser158AspGly152Asp＋Asn153Glu＋Ile159Thr＋Ser160GluGly152Glu＋Asn153Asp＋Ile159Leu＋Ser160GluAla156Thr＋Ser158Asp＋Ser160Glu＋Tyr161CysSer158Glu＋Ile159Met＋Ser160Asp＋Tyr161ProSer158Glu＋Ile159Cys＋Ser160Glu＋Tyr161GlyGly155Pro＋Ser158Glu＋Ile159Gln＋Ser160AspGly157Pro＋Ser158Asp＋Ser160Glu＋Tyr161MetGly152Glu＋Ser154Glu＋Gly157Asn＋Ser160GluGly152Glu＋Ser154Asp＋Gly157Gln＋Ser160AspGly152Glu＋Ser154Asp＋Gly1575er＋Ser160GluGly152Glu＋Ala156Gly＋Ser158Asp＋Ser160GluGly152Asp＋Ser158Asp＋Ile159Leu＋Ser160GluGly152Asp＋Gly155Gln＋Ser158Asp＋Ser160GluAla156Glu＋Ser158Glu＋Ser160Asp＋Tyr161GlyGly157Asp＋Ser158Glu＋Ile159His＋Ser160GluGly152Asn＋Ser154Glu＋Gly155Asp＋Ser158AspAsn153Glu＋Ser154Asp＋Gly157Pro＋Ser160GluAsn153Asp＋Ser154Asp＋Ile159Ser＋Ser160GluAsn153Asp＋Ser154Asp＋Ser160Asp＋Tyr161IleGly152Asn＋Ser154Glu＋Gly157Glu＋Ser158AspGly152Glu＋Ala156Asp＋Ser158Glu＋Ile159ValAsn153Asp＋Ser154Glu＋Ala156Ser＋Gly157AspAsn153Glu＋Ser154Glu＋Gly155Ser＋Gly157AspSer154Glu＋Gly155Asp＋Gly157Pro＋Ser160AspGly152Asp＋Asn153Glu＋Ser158Glu＋Ile159Ala表19第5环状区-单突变的突变体Alal81AsnAla181AspAla181GlnAla181GluAla181GlyAla181HisAla181ProAla181SerAla181ThrSer182AspSer182GluPhe183AlaPhe183AsnPhe183AspPhe183CysPhe183GlnPhe183GluPhe183GlyPhe183HisPhe183IlePhe183LeuPhe183MetPhe183ProPhe183SerPhe183ThrPhe183TyrPhe183ValSer184AspSer184GluGln185AsnGln185AspGln185GluGln185Ser表20第5环状区-双突变的突变体Ala181Asp＋Phe183GlnSer182Asp＋Gln185AsnPhe183Met＋Gln185GluSer182Glu＋Gln185AsnAla181Pro＋Ser182GluAla181Asn＋Gln185GluSer182Glu＋Phe183LeuAla181Pro＋Gln185AspPhe183Ser＋Gln185GluAla181Gln＋Ser182GluPhe183Ile＋Gln185GluAla181Ser＋Gln185AspAla181Gln＋Phe183SerAla181Thr＋Phe183AsnAla181Gly＋Gln185AspAla181His＋Ser182GluPhe183Gln＋Gln185SerPhe183Pro＋Gln185SerAla181Asn＋Ser182AspAla181Ser＋Gln185GluAla181Asn＋Phe183AsnAla181His＋Phe183AspAla181Asn＋Ser182GluSer182Asp＋Phe183HisAla181Asp＋Phe183IlePhe183Leu＋Gln185AspSer182Glu＋Phe183SerAla181His＋Gln185AspPhe183Val＋Gln185AspSer182Asp＋Phe183ThrPhe183Gly＋Gln185AspAla181Thr＋Gln185GluAla181His＋Gln185GluSer182Asp＋Phe183ValAla181Asp＋Phe183SerSer182Asp＋Phe183CysAla181Pro＋Phe183AspAla181Gly＋Phe183AlaAla181Pro＋Phe183AsnPhe183Pro＋Gln185AspSer182Glu＋Phe183TyrPhe183Cys＋Gln185SerAla181Thr＋Ser182GluAla181Pro＋Phe183AlaSer182Glu＋Phe183ThrAla181Pro＋Phe183ProSer182Glu＋Gln185SerAla181Gln＋Ser182AspPhe183Gly＋Gln185GluAla181Gln＋Gln185AspPhe183Ser＋Gln185AspSer182Asp＋Phe183MetPhe183Ala＋Gln185AsnAla181Asp＋Phe183HisPhe183Val＋Gln185GluAla181Ser＋Ser182GluAla181Asn＋Phe183ProPhe183Tyr＋Gln185GluAla181Glu＋Phe183CysSer182Glu＋Phe183Ile表21第5环状区-三突变的突变体Ala181Glu＋Phe183Val＋Gln185SerAla181Pro＋Ser182Asp＋Gln185AsnAla181Thr＋Phe183His＋Gln185AspSer182Glu＋Phe183Ala＋Gln185AsnAla181Thr＋Ser182Asp＋Gln185AsnAla181Gln＋Ser182Asp＋Phe183AlaSer182Asp＋Phe183Asn＋Gln185AsnAla181Gln＋Phe183Gln＋Gln185GluSer182Asp＋Phe183Met＋Gln185AsnAla181Pro＋Ser182Glu＋Phe183GlyAla181His＋Phe183Ala＋Gln185SerAla181Ser＋Ser182Glu＋Gln185AsnAla181Gly＋Phe183Ile＋Gln185GluAla181Asn＋Phe183Tyr＋Gln185AsnSer182Asp＋Phe183Leu＋Gln185AsnSer182Asp＋Phe183Val＋Gln185AsnAla181Glu＋Phe183Leu＋Gln185SerAla181Glu＋Phe183Tyr＋Gln185SerAla181Gln＋Ser182Asp＋Phe183GlyAla181Thr＋Phe183Ile＋Gln185SerAla181Gln＋Ser182Glu＋Gln185SerAla181Gln＋Ser182Glu＋Phe183HisAla181Asp＋Phe183Ala＋Gln185AsnAla181Thr＋Ser182Asp＋Phe183ThrAla181Ser＋Phe183Pro＋Gln185GluSer182Glu＋Phe183Ala＋Gln185SerAla181Asn＋Phe183Cys＋Gln185SerAla181Pro＋Phe183Thr＋Gln185AspAla181Thr＋Phe183Val＋Gln185AsnAla181Gln＋Ser182Asp＋Gln185SerAla181Asn＋Ser182Asp＋Gln185SerSer182Asp＋Phe183Thr＋Gln185SerAla181Ser＋Phe183Asp＋Gln185AsnSer182Asp＋Phe183Thr＋Gln185AsnAla181Gly＋Phe183Asn＋Gln185SerAla181Asp＋Phe183Met＋Gln185SerAla181Asp＋Phe183Pro＋Gln185AsnAla181Gly＋Ser182Asp＋Phe183GlyAla181Gln＋Ser182Glu＋Gln185AsnAla181Thr＋Ser182Asp＋Gln185SerAla181Asn＋Phe183Asn＋Gln185AspAla181Thr＋Phe183Asp＋Gln185SerAla181Gln＋Phe183Ser＋Gln185SerSer182Asp＋Phe183Gly＋Gln185SerAla181Gly＋Ser182Glu＋Phe183LeuAla181Asn＋Ser182Glu＋Gln185AsnAla181Glu＋Ser182Glu＋Phe183SerAlal81Glu＋Ser182Glu＋Phe183GlyAla181Glu＋Ser182Glu＋Phe183LeuAla181Glu＋Ser182Glu＋Phe183ValAla181Glu＋Ser182Asp＋Phe183MetAla181Asp＋Ser182Asp＋Phe183MetAla181Asp＋Ser182Asp＋Phe183CysAla181Glu＋Ser182Asp＋Gln185AsnAla181Asp＋Ser182Asp＋Gln185AsnAla181Glu＋Ser182Glu＋Phe183AsnAla181Asp＋Ser182Glu＋Phe183SerAla181Glu＋Ser182Asp＋Gln1855erAla181Glu＋Ser182Glu＋Phe183ThrAla181Glu＋Ser182Asp＋Phe183Leu
表22第5环状区-四突变的突变体Ala181Ser＋Ser182Asp＋Phe183Ala＋Gln185SerAla181Ser＋Ser182Glu＋Phe183Pro＋Gln185SerAla181Gly＋Ser182Asp＋Phe183Gly＋Gln185AsnAla181Gly＋Ser182Glu＋Phe183Pro＋Gln18SSerAla181Thr＋Ser182Glu＋Phe183Thr＋Gln185SerAla181Gly＋Ser182Glu＋Phe183Thr＋Gln185SerAla181His＋Ser182Glu＋Phe183His＋Gln185AsnAla181Gly＋Ser182Glu＋Phe183Ser＋Gln185SerAla181Gln＋Ser182Glu＋Phe183His＋Gln185AsnAla181Asn＋Ser182Glu＋Phe183Cys＋Gln185SerAla181Pro＋Ser182Glu＋Phe183Met＋Gln185SerAla181Gln＋Ser182Asp＋Phe183Gln＋Gln185AsnAla181Thr＋Ser182Asp＋Phe183His＋Gln185AsnAla181Gly＋Ser182Glu＋Phe183Leu＋Gln185SerAla181Asn＋Ser182Asp＋Phe183Ile＋Gln185SerAla181Gly＋Ser182Asp＋Phe183Ala＋Gln185SerAla181Asn＋Ser182Asp＋Phe183Ser＋Gln185AsnAla181Asn＋Ser182Glu＋Phe183Gly＋Gln185SerAla181Asn＋Ser182Glu＋Phe183Met＋Gln185AsnAla181His＋Ser182Glu＋Phe183Asn＋Gln185AsnAla181Gly＋Ser182Glu＋Phe183Tyr＋Gln185AsnAla181Asn＋Ser182Asp＋Phe183Asn＋Gln185AsnAla181Glu＋Ser182Glu＋Phe183Asn＋Gln185AsnAla181Asp＋Ser182Glu＋Phe183Ile＋Gln185AsnAla181Glu＋Ser182Asp＋Phe183Gln＋Gln185AsnAla181Asp＋Ser182Asp＋Phe183His＋Gln185AsnAla181Glu＋Ser182Asp＋Phe183Ser＋Gln185AsnAla181Glu＋Ser182Asp＋Phe183Leu＋Gln185AsnAla181Glu＋Ser182Asp＋Phe183Leu＋Gln185SerAla181Asp＋Ser182Glu＋Phe183Thr＋Gln185SerAla181Asp＋Ser182Asp＋Phe183Gln＋Gln185AsnAla181Asp＋Ser182Asp＋Phe183Ala＋Gln185AsnAla181Asp＋Ser182Glu＋Phe183Ala＋Gln185AsnAla181G1u＋Ser182Asp＋Phe183Met＋Gln185SerAla181Glu＋Ser182Glu＋Phe183Thr＋Gln185AsnAla181Asp＋Ser182Asp＋Phe183Gly＋Gln185SerAla181Glu＋Ser182Asp＋Phe183Ala＋Gln185AsnAla181His＋Ser182Glu＋Phe183Glu＋Gln185AsnAla181Ser＋Ser182Asp＋Phe183Glu＋Gln185SerAla18lSer＋Ser182Asp＋Phe183Asp＋Gln185SerAla181Pro＋Ser182Glu＋Phe183Asp＋Gln185SerAla181Asp＋Ser182Asp＋Phe183Asp＋Gln185AsnAla181Glu＋Ser182Asp＋Phe183Glu＋Gln185SerAla181Ser＋Ser182Asp＋Phe183Glu＋Gln185GluAla181Gly＋Ser182Glu＋Phe183Glu＋Gln185GluAla181Gln＋Ser182Glu＋Phe183Asp＋Gln185AspAla181Ser＋Ser182Asp＋Phe183Asp＋Gln185AspAla181Thr＋Ser182Asp＋Phe183Asp＋Gln185AspAla181Pro＋Ser182Asp＋Phe183Glu＋Gln185AspAla181Asn＋Ser182Glu＋Phe183Asp＋Gln185AspAla181Thr＋Ser182Asp＋Phe183Asp＋Gln185GluAla181Pro＋Ser182Glu＋Phe183Glu＋Gln185GluAla181Asn＋Ser182Asp＋Phe183Asp＋Gln185GluAla181Thr＋Ser182Glu＋Phe183Asp＋Gln185AspAla181His＋Ser182Glu＋Phe183Glu＋Gln185GluAla181Glu＋Ser182Glu＋Phe183Ser＋Gln185GluAla181Glu＋Ser182Glu＋Phe183Val＋Gln185GluAla181Asp＋Ser182Glu＋Phe183Pro＋Gln185GluAla181Glu＋Ser182Glu＋Phe183Thr＋Gln185AspAla181Asp＋Ser182Asp＋Phe183Cys＋Gln185Glu表23第6环状区-单突变的突变体Val193AlaVal193AsnVal193AspVal193CysVal193GlnVal193GluVal193GlyVal193HisVal193MetVal193ProVal193SerVal193ThrAla194AsnAla194AspAla194GlnAla194GluAla194GlyAla194HisAla194ProAla194SerAla194ThrPro195AsnPro195AspPro195GlnPro195GluPro195GlyPro195SerGly196AsnGly196AspGly196GlnGly196GluGly196ProGly196SerVal197AlaVal197AspVal197CysVal197GlnVal197GluVal197GlyVal197HisVal197MetVal197ProVal197SerVal199AlaVal199AsnVal199AspVal199CysVal199GlnVal199GluVal199GlyVal199HisVal199MetVal199ProVal199SerVal199ThrGln200AsnGln200SerSer201AspSer201GluThr202AsnThr202AspThr202GlnThr202GluThr202GlyThr202ProThr202SerTyr203AlaTyr203AsnTyr203AspTyr203GlnTyr203HisTyr203IleTyr203MetTyr203ProTyr203SerPro204AsnPro204AspPro204GlnPro204GluPro204GlyPro204SerGly205AsnGly205AspGly205GlnGly205GluGly205ProGly205SerSer206AspSer206GluThr207AsnThr207GlnThr207GluThr207GlyThr207ProThr207SerTyr208AlaTyr208AsnTyr208AspTyr208CysTyr208GlnTyr208GluTyr208GlyTyr208HisTyr208IleTyr208LeuTyr208MetTyr208ProTyr208ValAla209AsnAla209AspAla209GlnAla209GluAla209GlyAla209HisAla209ProAla209SerAla209ThrSer210AspSer210GluLeu211AlaLeu211AsnLeu211AspLeu211CysLeu211GlnLeu211GlyLeu211HisLeu211IleLeu211MetLeu211ProLeu211SerLeu211ThrLeu211ValAsn212GluGly213AsnGly213AspGly213GlnGly213GluGly213ProGly213SerThr214AsnThr214AspThr214GlnThr214GluThr214GlyThr214ProThr214Ser
表24第6环状区-双突变的突变体Gly196Asn＋Ala209AspVal199Ser＋Tyr208AsnPro195Asn＋Thr207AspVal197Ala＋Asn212AspAsn198Gln＋Thr207SerVal199Gly＋Tyr208IleGly196Asn＋Ala209ProVal199Gly＋Gln200SerPro195Ser＋Gly205GlnVal199Ser＋Leu211AspGly196Gln＋Val197AsnThr202Asn＋Thr214GluAla194Thr＋Thr202ProVal199Asn＋Ser210GluThr202Ser＋Asn212GlnSer210Asp＋Thr214AsnPro195Gly＋Asn212AspAsn198Gln＋Tyr208GlnVal197Asn＋Thr214GluTyr208Asp＋Leu211GlnThr202Ser＋Ser210GluVal197Cys＋Ser210GluGln200Glu＋Tyr203GlyAla209Thr＋Leu211GluVal197Ser＋Gln200GluThr202Gly＋Ser210GluGln200Ser＋Gly213ProGly196Ser＋Thr207GlyPro204Gln＋Ser210GluVal199Cys＋Pro204GlyGly213Pro＋Thr214ProGly196Asn＋Ser210GluThr207Gln＋Ser210GluVal199Met＋Gln200AspThr202Ser＋Tyr203IlePro195Ser＋Asn212SerVal197Glu＋Tyr208GlnAsn198Glu＋Leu211CysGly196Gln＋Ser206GluAla194Ser＋Thr214SerVal197Ser＋Tyr203AlaSer210Asp＋Gly213ProTyr203His＋Gly213AspVal197Ser＋Val199SerAla209Pro＋Ser210AspThr207Asp＋Thr214SerThr207Gly＋Ala209HisPro195Asn＋Asn198AspVal197Pro＋Ser206AspGln200Ser＋Asn212AspVal197Thr＋Asn212AspGln200Asn＋Thr214GluAla209Asp＋Thr214SerAla209Asn＋Asn212SerAla194Ser＋Asn212GluVal197Glu＋Gly213AsnTyr203Cys＋Asn212AspPro195Gln＋Val197CysAsn198Ser＋Tyr208AlaGly205Gln＋Ala209GluGly205Ser＋Leu211SerGly205Gln＋Tyr208AlaAla194Ser＋Asn198AspAla194Thr＋Pro195SerVal199Thr＋Pro204GluThr202Gln＋Ser210AspSer206Glu＋Leu21lHisAsn198Glu＋Thr202AsnSer210Asp＋Gly213SerGly196Ser＋Thr202GlnPro204Gln＋Gly213AspAsn198Glu＋Pro204GlyThr202Gly＋Tyr208MetPro195Gly＋Gly213GluPro195Ser＋Pro204AspPro195Ser＋Tyr203ThrAla194Asn＋Tyr203ProAla194Pro＋Gln200AspPro204Asp＋Gly205AsnGly196Pro＋Asn198GlnThr207Ser＋Leu211GlyPro195Ser＋Leu211IlePro204Glu＋Leu211GlyVal199Ser＋Tyr208AlaGly196Ser＋Ser210AspAla194Thr＋Leu211GlyThr207Ser＋Ser210GluGly196Pro＋Thr207GluPro204Glu＋Leu211MetPro195Asn＋Asn198SerTyr203Met＋Thr207GlnPro204Gly＋Ala209GluVal197Glu＋Asn212SerPro1955er＋Ala209ProGly196Ser＋Leu211GlyGly205Pro＋Leu211MetVal197Gln＋Pro204SerVal199Asn＋Ser206AspVal197Asp＋Thr202AsnGly196Gln＋Thr214GlyPro204Asn＋Asn212GlnVal199Asn＋Gln200AsnPro195Gly＋Leu211GlyTyr203Cys＋Pro204SerGln200Asp＋Leu211CysAsn198Gln＋Ser210GluAla194Gly＋Gln200GluGly196Gln＋Pro204GluVal197Pro＋Thr202AsnAla194Gly＋Val199AlaAla194Thr＋Leu211AsnAsn198Gln＋Gly205AspTyr203Asn＋Leu211HisAsn212Asp＋Gly213GlnAsn198Gln＋Ser206AspVal199Thr＋Gln200GluThr202Ser＋Asn212AspAla194Gly＋Ser210GluSer206Glu＋Ala209AsnAla209Asp＋Asn212SerVal199Gly＋Asn212SerAla194Pro＋Gly213AspGln200Asp＋Pro204GlyVal197Thr＋Tyr203AlaAsn198Gln＋Leu211ProPro195Gln＋Leu211GlnGly196Gln＋Thr214GluPro195Asn＋Ser210GluThr207Asn＋Leu211AspSer210Glu＋Gly213ProGly205Pro＋Thr207AspGln200Glu＋Thr202GlnThr202Ser＋Gly213AspPro204Gln＋Thr214GlnGly196Ser＋Gln200AspVal197Ser＋Gln200AspThr202Pro＋Thr207AspTyr203Thr＋Tyr208HisPro195Gln＋Thr202AsnVal197Asn＋Asn212AspVal197Met＋Thr202GlnVal199Met＋Thr202ProAla209Glu＋Leu21lMetGly196Asn＋Val199HisAla194Gly＋Ser206GluGly196Asn＋Pro204AsnGly196Asn＋Gln200GluVal197Ser＋Ser210AspPro195Gln＋Pro204GlnAla194His＋Asn198GlnVal197Cys＋Asn212GlnAsn198Glu＋Ala209SerAla194Gln＋Val199ProAsn198Asp＋Val199GlnAsn198Asp＋Thr214ProPro204Asn＋Tyr208AspTyr203Ser＋Tyr208LeuGly196Gln＋Ala209AspSer206Glu＋Asn212GlnThr207Ser＋Ala209HisVal197Thr＋Ser210AspTyr208Gln＋Ser210AspAsn198Ser＋Ser210GluGly196Ser＋Thr214AspAla194Gln＋Thr202AsnPro195Gly＋Thr214AsnLeu21lThr＋Thr214AspPro195Ser＋Asn198GluGly196Ser＋Gly205SerAla194His＋Thr202AsnThr207Gly＋Tyr208MetAla194Ser＋Val197His表25第6环状区-三突变的突变体Ala194Thr＋Ser206Glu＋Tyr208CysGly196Asn＋Tyr203Pro＋Ala209GlyPro195Gly＋Asn198Ser＋Thr207GluPro195Gly＋Gly205Gln＋Gly213SerTyr203Leu＋Pro204Gln＋Gly205GlnGly196Gln＋Val197Asn＋Ser206AspLeu21lSer＋Gly213Ser＋Thr214GlnPro195Asn＋Pro204Glu＋Gly205GlnAsn198Asp＋Thr202Gly＋Gly213GlnTyr203Ala＋Leu211His＋Thr214GlnAla194Thr＋Val197Ser＋Val199GlyAla194Gln＋Thr207Asp＋Thr214SerGly196Asn＋Pro204Gln＋Ala209AsnThr202Pro＋Tyr208Asp＋Leu21lThrVal197Gly＋Ala209Gln＋Asn212GlnGly196Ser＋Gly205Asp＋Leu21lIleAla194Ser＋Val197Pro＋Asn198AspGly196Asn＋Asn198Gln＋Leu211SerAla194Thr＋Ala209His＋Thr214AspAla194Pro＋Pro195Ser＋Asn212AspThr202Ser＋Tyr203Gln＋Ser210AspThr202Ser＋Tyr203Leu＋Thr214GluAla194Ser＋Pro195Gly＋Leu211ValAla194Pro＋Thr207Glu＋Leu21lAlaGly196Asn＋Ser206Glu＋Leu21lGlyGly196Ser＋Gly205Ser＋Ser210AspAla194Thr＋Val197Gly＋Leu211SerPro204Glu＋Ala209His＋Thr214ProGly196Ser＋Gly205Asp＋Leu211HisVal197Cys＋Thr202Gly＋Thr207GlnAsn198Ser＋Pro204Gly＋Gly213AspAla194Gly＋Ala209Pro＋Ser210AspGly196Pro＋Tyr203Val＋Tyr208CysVal199Ala＋Thr207Asn＋Tyr208ValTyr203Cys＋Ser210Asp＋Leu211ProAla1945er＋Val1975er＋Gln200AspAla194Asn＋Pro195Gly＋Leu211IleVal199Ser＋Leu211Asn＋Gly213AspAla194Asn＋Ser206Glu＋Thr214GlyAla194Gly＋Pro195Gly＋Val197AspPro195Gln＋Ala209Ser＋Ser210AspVal197Thr＋Asn198Gln＋Tyr203LeuVal197Ala＋Thr202Gln＋Ser206GluGly196Pro＋Pro204Gly＋Asn212GluThr202Gly＋Pro204Glu＋Tyr208SerGln200Ser＋Thr202Pro＋Pro204SerGln200Glu＋Tyr203Thr＋Pro204AsnAsn198Asp＋Val199Gln＋Pro204AsnAla194Pro＋Pro195Gln＋Val199GlyPro195Asn＋Pro204Asn＋Ser206GluThr202Asn＋Leu211Thr＋Asn212GlnGly196Gln＋Thr202Pro＋Gly213AspAsn198Glu＋Gln200Asn＋Leu211GlyVal199Gly＋Thr202Gln＋Gly213GluTyr203Ser＋Thr207Pro＋Ser210AspPro195Gly＋Gly196Gln＋Val197GluPro195Gly＋Thr207Gly＋Tyr208SerThr202Gly＋Leu211Glu＋Gly213GlnVal197Gly＋Thr202Pro＋Asn212GluGly196Gln＋Gly205Gln＋Ser206GluPro195Gly＋Val199Thr＋Thr214GluGly196Ser＋Gln200Ser＋Leu211AspGly196Asn＋Tyr208Glu＋Thr214ProTyr203Thr＋Pro204Asn＋Ser206GluVal199Thr＋Thr202Asn＋Tyr208GlyThr202Pro＋Tyr203Thr＋Pro204GluGly196Gln＋Asn212Asp＋Thr214GlnPro195Gly＋Thr202Gly＋Asn212SerPro195Gly＋Tyr208Gln＋Ser210AspAlal94Pro＋Ser206Glu＋Leu211SerAla194His＋Pro204Asn＋Gly213GlnGly196Ser＋Tyr208Met＋Ser210AspVal197Glu＋Val199Ser＋Tyr203AsnThr202Gly＋Ala209Asn＋Asn212AspVal197Ser＋Ser210Asp＋Gly213ProAla194Ser＋Tyr208Ile＋Asn212GluAla194Gly＋Asn198Gln＋Ala209HisAla194Ser＋Thr207Pro＋Gly213GluVal199Thr＋Leu211Asp＋Asn212SerVal199Pro＋Thr202Ser＋Leu211AsnSer206Glu＋Tyr208Cys＋Thr214AsnAla209ser＋Leu211Ile＋Asn212GluAsn198Glu＋Tyr208Ala＋Ala209SerAla194Pro＋Asn198Asp＋Gly213GlnAla194Asn＋Pro195Ser＋Leu211GluGly196Pro＋Pro204Gly＋Gly205ProTyr203Ile＋Tyr208Thr＋Ala209GluGly196Gln＋Thr202Gln＋Asn212AspAla194Asn＋Val197Pro＋Thr202ProAla194Gln＋Val199Gly＋Ala209AspAla194Ser＋Pro195Gly＋Asn212GlnPro195Gln＋Tyr203Pro＋Ser210AspVal197Thr＋Thr202Gln＋Tyr208AsnAla194Thr＋Gly196Ser＋Leu211AsnAla194Gly＋Thr202Ser＋Pro204AspAla194Gly＋Gly205Glu＋Gly213SerVal199Pro＋Thr207Pro＋Tyr208MetAla194Pro＋Asn198Glu＋Ala209HisTyr208Leu＋Ser210Glu＋Thr214ProGly196Asn＋Val199Ala＋Gly205AspGly196Pro＋Gly205Glu＋Tyr208SerPro195Asn＋Gly196Gln＋Val197CysAsn198Ser＋Gln200Glu＋Leu211AsnThr202Ser＋Tyr203Cys＋Thr214GlnAla194Ser＋Gly196Gln＋Thr207AspGly196Ser＋Gly205Glu＋Tyr208IleTyr203Met＋Gly205Asn＋Gly213ProPro204Gln＋Gly205Asp＋Ala209HisGly205Asn＋Leu211Asp＋Thr214SerTyr203Met＋Ala209His＋Leu211MetVal199Asn＋Thr202Ser＋Gly213GluVal197Thr＋Pro204Asp＋Asn212GlnAla194Gly＋Ser210Glu＋Leu211ProVal197His＋Ala209Gly＋Ser210AspGln200Glu＋Thr202Gln＋Tyr208HisAla194Ser＋Ser206Asp＋Leu211AlaPro195Ser＋Asn198Asp＋Gly205ProTyr203Pro＋Thr207Asp＋Thr214ProPro195Ser＋Val199His＋Leu211ProTyr203Met＋Thr207Glu＋Ala209HisVal197Cys＋Gln200Asn＋Ser206GluVal197Glu＋Thr202Gln＋Tyr208HisPro195Gln＋Val197Cys＋Pro204GlyVal199Pro＋Gly205Ser＋Gly213AspAla194Thr＋Thr207Ser＋Ala209GluVal197Met＋Gly205Pro＋Tyr208GlyGly196Ser＋Gly205Glu＋Ala209ThrPro195Ser＋Val199Thr＋Gly205SerAla194Ser＋Asn198Asp＋Gly213SerAsn198Glu＋Val199Ser＋Thr207ProAla194Thr＋Gly196Pro＋Ser210GluGly196Asn＋Thr202Pro＋Leu21lMetTyr203Met＋Asn212Asp＋Thr214SerTyr203Gly＋Ala209Ser＋Ser210AspAla194Ser＋Thr207Ser＋Leu211ProTyr208Cys＋Gly213Asn＋Thr214ProVal197Met＋Pro204Glu＋Gly213SerGln200Ser＋Tyr203Ala＋Pro204AspPro195Gly＋Ser206Asp＋Leu211IleTyr203Gly＋Tyr208Asp＋Leu211AsnVal197Thr＋Ser206Glu＋Thr214ProTyr203Ala＋Pro204Asn＋Gly213GluTyr208Ile＋Ala209Gly＋Leu21lGlnAsn198Ser＋Pro204Gly＋Asn212GluAla194Gln＋Val197Cys＋Pro204AsnAla209Thr＋Leu211Ser＋Gly213ProGly196Ser＋Thr202Pro＋Asn212AspVal197His＋Tyr203Gln＋Tyr208CysVal199Ser＋Leu211Thr＋Gly213ProPro195Asn＋Val197Asn＋Thr202ProVal199Pro＋Pro204Gly＋Gly205AsnPro195Gly＋Asn198Asp＋Thr214GlyGln200Glu＋Tyr203Ser＋Asn212GlnPro195Gly＋Val199Gln＋Thr214ProThr202Ser＋Pro204Ser＋Gly213GluPro195Gly＋Thr202Pro＋Gly205GluThr202Gln＋Tyr203Gly＋Ala209AsnPro195Asn＋Gly196Gln＋Asn212AspThr202Gly＋Tyr208Ile＋Leu211ValAsn198Glu＋Tyr203Cys＋Pro204GlyAla194Ser＋Thr207Ser＋Ser210AspGln200Asp＋Thr202Pro＋Tyr203AlaVal199Cys＋Ala209Gln＋Ser210AspAla194Asn＋Val199Cys＋Leu211ProGln200Ser＋Ser210Glu＋Leu211AsnTyr203Gln＋Thr207Glu＋Tyr208SerAla194Thr＋Thr202Asn＋Thr207GlyPro195Gly＋Gly196Gln＋Ser210AspGln200Ser＋Pro204Gly＋Ser206GluAla194Asn＋Gly196Gln＋Asn198GluVal197Thr＋Pro204Gln＋Gly205GluVal199Ala＋Gln200Asp＋Gly213AsnVal197Met＋Tyr203Val＋Gly213ProAla194Gln＋Tyr203Val＋Thr214GluVal199Cys＋Ala209His＋Leu211MetThr202Gln＋Tyr203Ile＋Ala209GlnAla194Gly＋Ser206Glu＋Leu211GlyGly196Pro＋Tyr208Gly＋Ser210GluVal197Gln＋Thr202Gly＋Gly205AspVal197Pro＋Gly205Asp＋Thr214Pro表26第6环状区-四突变的突变体Pro195Asn＋Val197Ala＋Thr207Ser＋Asn212AspAsn198Gln＋Val199Gly＋Tyr208Ile＋Asn212AspTyr203His＋Leu211Pro＋Asn212Gln＋Gly213GlnAla194His＋Asn198Ser＋Thr202Asn＋Ala209AspGly196Gln＋Asn198Ser＋Thr202Ser＋Pro204GluPro195Gln＋Asn198Gln＋Gly205Glu＋Ala209ProVal199His＋Pro204Asp＋Gly205Pro＋Leu211GlyGly196Gln＋Gly205Asn＋Ser206Asp＋Thr214SerGln200Glu＋Thr202Pro＋Leu211Asn＋Gly213AsnGly196Gln＋Pro204Asn＋Asn212Gln＋Thr214GlyPro195Gly＋Val199Asn＋Gln200Asn＋Leu211GlyPro195Gln＋Pro204Ser＋Gly205Glu＋Tyr208IleVal199Ala＋Tyr203Gln＋Thr207Gly＋Ser210GluAla194His＋Val199Ser＋Pro204Asn＋Gly205GluGly196Pro＋Asn198Asp＋Gln200Asn＋Ala209ProPro195Gln＋Gln200Asp＋Tyr203Asn＋Leu211CysPro195Ser＋Thr202Pro＋Asn212Asp＋Thr214SerAla194Gly＋Val197Glu＋Tyr203Leu＋Leu211CysAsnl98Gln＋Thr202Pro＋Tyr208Met＋Thr214GluGly196Ser＋Val199Ser＋Gly205Gln＋Ser210GluTyr203Ala＋Gly205Gln＋Ser206Asp＋Leu211GlnGly196Ser＋Va1199His＋Gln200Asp＋Thr207SerPro195Gln＋Gly196Gln＋Gln200Glu＋Tyr203SerGly196Ser＋Gln200Glu＋Gly205Gln＋Gly213AsnPro195Ser＋Glyl96Asn＋Ser206Glu＋Thr214GlyGly196Asn＋Val199Asn＋Ser206Glu＋Tyr208ProVal197Gln＋Gly205Ser＋Ser210Glu＋Thr214SerPro195Asn＋Gln200Glu＋Gly205Asn＋Leu211ProTyr203Pro＋Tyr208His＋Leu211Val＋Thr214ProAla194His＋Val199Ser＋Thr202Gly＋Tyr208AspPro195Asn＋Val199Asn＋Ser206Asp＋Gly213SerAla194Ser＋Pro195Asn＋Val199Ala＋Gly205GluAla194Thr＋Gly196Gln＋Val199Gly＋Tyr203IleVal199Ala＋Ala209Asp＋Leu211Val＋Asn212GlnVal197Gly＋Asn198Ser＋Tyr203Leu＋Ser210AspGln200Ser＋Thr207Gly＋Tyr208Thr＋Ser210AspPro204Gly＋Gly205Asp＋Thr207Asn＋Tyr208LeuAla194Gln＋Thr202Ser＋Ala209Ser＋Asn212GluVal197Met＋Thr202Gly＋Pro204Gly＋Thr214GluVal199Ala＋Pro204Gln＋Gly205Asp＋Tyr208GlyAsn198Glu＋Tyr208Cys＋Ala209Gln＋Leu211ValGln200Ser＋Pro204Ser＋Ala209Asn＋Leu211GluPro195Ser＋Val197Thr＋Gly205Gln＋Thr214ProPro195Asn＋Thr202Asn＋Gly205Asp＋Tyr208SerPro195Gly＋Tyr203Pro＋Leu211Ala＋Asn212GluThr202Gly＋Pro204Asn＋Gly205Asn＋Gly213GlnVal197Gly＋Gln200Glu＋Tyr208Ala＋Asn212SerAla194Ser＋Pro195Ser＋Tyr203Ala＋Ala209AspPro195Gln＋Val197Thr＋Val199Pro＋Ala209HisGly196Asn＋Ser206Asp＋Leu211His＋Thr214SerVal197Asp＋Tyr203His＋Tyr208Cys＋Thr214GlyAla194Gln＋Pro195Asn＋Ala209His＋Ser210AspVal199Ser＋Gln200Ser＋Tyr203Asn＋Ser206AspAla194Asn＋Pro195Gln＋Ser206Glu＋Gly213SerPro1955er＋Tyr203Met＋Ser206Asp＋Gly213GlnAla194Gly＋Val199Thr＋Tyr208His＋Gly213ProAla194Pro＋Val197Cys＋Gln200Asp＋Pro204GlyThr202Gln＋Thr207Glu＋Tyr208Cys＋Leu211ProGly196Gln＋Asn198Glu＋Tyr203Gln＋Gly205ProThr202Asn＋Tyr203Pro＋Gly205Ser＋Gly213GluPro195Asn＋Asn198Ser＋Ala209Gly＋Leu211ThrAsn198Asp＋Gln200Ser＋Tyr203Pro＋Gly205AsnPro195Gln＋Val199Gly＋Tyr203Asn＋Thr207GluAla194Gln＋Thr202Gln＋Tyr203His＋Tyr208HisAla194Ser＋Pro204Asp＋Thr207Pro＋Tyr208ThrAsn198Gln＋Val199Gln＋Gln200Asn＋Ala209GlnAla194Gly＋Asn198Ser＋Pro204Asp＋Ala209HisGly196Gln＋Val197Thr＋Ser206Glu＋Ala209ProGln200Ser＋Thr202Pro＋Tyr203Thr＋Thr207GlyAla194Thr＋Val197Gly＋Gln200Asn＋Thr207AspVal197Pro＋Val199Asn＋Gln200Glu＋Leu211IleGly196Pro＋Pro204Ser＋Ala209Ser＋Leu21lGlnPro195Gly＋Gly196Gln＋Asn198Gln＋Tyr208GluGln200Ser＋Tyr203Met＋Gly205Glu＋Tyr208AsnAla194Gln＋Pro195Gly＋Ala209Gly＋Ser210GluGly196Asn＋Val197Asn＋Ser210Asp＋Asn212GlnGly196Gln＋Thr202Ser＋Gly205Ser＋Ser210GluGln200Asn＋Gly205Pro＋Leu211Cys＋Thr214ProAla194Ser＋Gly196Gln＋Thr207Ser＋Ser210AspVal197Gln＋Thr202Pro＋Tyr203Ala＋Ser206AspGly196Gln＋Asn198Glu＋Asn212Ser＋Gly213AsnPro195Asn＋Val197Gln＋Gly205Gln＋Leu21lGluThr207Glu＋Tyr208Leu＋Leu211Met＋Gly213GlnPro195Ser＋Val197Pro＋Pro204Glu＋Leu21lCysPro195Gly＋Gly196Ser＋Val199Gln＋Thr214AspVal197Asn＋Gly205Asn＋Thr207Ser＋Ser210AspAla194His＋Tyr203His＋Gly205Pro＋Ser206GluPro195Asn＋Asn198Gln＋Val199Gln＋Leu211GlnVal197Pro＋Thr202Gly＋Pro204Gly＋Thr207SerThr202Pro＋Tyr203Leu＋Leu211Ala＋Thr214AspPro195Asn＋Val199His＋Tyr203Met＋Thr207GluVal199Cys＋Pro204Gly＋Thr207Ser＋Thr214AspAla194Ser＋Pro195Gln＋Tyr203Thr＋Tyr208ProVa1199Cys＋Tyr203Cys＋Ser206Asp＋Tyr208AlaVa1197Thr＋Thr202Gln＋Leu211Glu＋Asn212SerGly196Ser＋Val199Pro＋Ala209Gln＋Leu211AsnAla194Gly＋Ala209Gln＋Asn212Asp＋Thr214GlnAla194Gln＋Thr207Asn＋Ala209Pro＋Leu211HisGly196Ser＋Val197Ser＋Thr202Gln＋Leu211GlnAla194Thr＋Val197Ser＋Thr202Pro＋Thr207ProGly196Pro＋Thr202Pro＋Gly205Glu＋Gly213ProAla194Pro＋Pro195Gly＋Val197Asn＋Val199CysThr202Gly＋Thr207Ser＋Tyr208Pro＋Leu211ProThr202Asn＋Pro204Asp＋Thr207Ser＋Gly213AsnGly196Gln＋Val197Cys＋Tyr203Met＋Ala209AsnGly196Gln＋Ser206Glu＋Thr207Ser＋Gly213ProPro195Ser＋Val199Cys＋Ser210Glu＋Asn212SerAla194Ser＋Ser206Asp＋Asn212Ser＋Thr214GlnGly196Gln＋Gln200Ser＋Thr202Gln＋Ala209SerGly196Gln＋Tyr203Thr＋Thr207Gln＋Asn212GluAla194Pro＋Thr202Gln＋Tyr208Cys＋Leu211ProGln200Asp＋Tyr203Cys＋Leu211Gln＋Asn212GlnGly196Asn＋Tyr203Met＋Leu211Asn＋Gly213AsnPro195Gln＋Gly196Asn＋Gln200Ser＋Thr202GlnThr202Pro＋Tyr203Gly＋Asn212Ser＋Thr214GlyGln200Asp＋Tyr203Thr＋Pro204Gln＋Gly205SerGly196Ser＋Thr202Pro＋Pro204Asn＋Ala209SerGly196Ser＋Gln200Ser＋Pro204Gly＋Ser210AspAla194Gln＋Gln200Asn＋Thr202Ser＋Gly205GlnPro195Gly＋Gln200Asn＋Thr207Asp＋Ala209HisGln200Asn＋Thr202Ser＋Tyr203Ala＋Thr214GlyGln200Glu＋Ala209Pro＋Leu211Ile＋Gly213AsnAla194Asn＋Thr202Ser＋Tyr203Met＋Ser206GluVal197Asn＋Asn198Ser＋Thr207Gln＋Thr214GluVal197Met＋Gln200Ser＋Pro204Glu＋Ala209ProGly196Asn＋Val199Asn＋Leu211Glu＋Asn212GlnPro195Gln＋Thr202Gln＋Gly205Asp＋Tyr208AsnThr202Pro＋Ser206Asp＋Leu211Ile＋Gly213SerGly196Ser＋Gln200Glu＋Thr202Gln＋Tyr208MetAsn198Glu＋Tyr203Thr＋Tyr208Asn＋Gly213GlnAsn198Ser＋Ser210Glu＋Leu211His＋Thr214ProGly196Ser＋Val197Gly＋Pro204Glu＋Thr214SerTyr203Met＋Ser206Glu＋Tyr208Ser＋Ala209HisGly196Gln＋Gln200Ser＋Leu21lCys＋Gly213ProAla194Gly＋Val199Asn＋Thr202Asn＋Gly213ProAsn198Ser＋Thr202Asn＋Tyr203Gly＋Ser210GluVal199Gln＋Gln200Ser＋Tyr203His＋Thr214AspThr202Gln＋Ser206Asp＋Thr207Ser＋Thr214GlyAla194Thr＋Tyr203Gln＋Thr207Asp＋Leu211IleAla209Ser＋Ser210Asp＋Asn212Gln＋Gly213SerVal197Ser＋Tyr203Gly＋Ser206Glu＋Asn212GlnAla194Ser＋Asn198Ser＋Gly205Gln＋Ser210AspAla194Gln＋Pro195Ser＋Val197Gln＋Pro204AsnAla194Gly＋Asn198Ser＋Ala209Glu＋Thr214AsnVal199Met＋Ser206Asp＋Gly213Asn＋Thr214SerThr202Gln＋Gly205Ser＋Leu211Asp＋Asn212GluProl95Ser＋Ala209Gly＋Gly213Asp＋Thr214AspThr202Gln＋Thr207Pro＋Gly213Glu＋Thr214AspAla194Ser＋Thr207Pro＋Ala209Glu＋Ser210GluGln200Asn＋Thr202Gln＋Ala209Glu＋Ser210GluGln200Ser＋Ala209Asp＋Ser210Asp＋Asn212GlnGly205Asn＋Ala209Asp＋Ser210Glu＋Asn212GlnPro204Gly＋Leu211Gln＋Asn212Glu＋Gly213AspTyr203Gly＋Pro204Asp＋Gly205Glu＋Leu211AlaThr202Ser＋Pro204Glu＋Gly205Asp＋Thr207GlnThr202Gly＋Pro204Asp＋Gly205Glu＋Gly213AsnVal199Gly＋Ser210Asp＋Leu211Asp＋Asn212GlnGly196Pro＋Thr202Ser＋Gly205Asp＋Ser206GluVal197Thr＋Gly205Asp＋Ser206Glu＋Gly213GlnPro204Asn＋Gly205Glu＋Ser206Glu＋Asn212SerAla194His＋Gly205Asp＋Ser206Asp＋Leu211HisPro195Asn＋Ser206Glu＋Thr207Asp＋Asn212SerTyr203Met＋Ser206Glu＋Thr207Glu＋Leu211GlyGly196Gln＋Pro204Asn＋Ser206Asp＋Thr207GluAla194Thr＋Val197Cys＋Ser206Asp＋Thr207AspAla194Pro＋Tyr208Asp＋Ala209Asp＋Gly213GlnGln200Asp＋Tyr208Thr＋Ala209Ser＋Ser210GluVal199Asn＋Gln200Asp＋Thr202Gln＋Ser210AspAla194Ser＋Gln200Glu＋Thr202Gly＋Ser210GluVal199Cys＋Gln200Asp＋Ala209Gln＋Ser210AspAla194Gln＋Val197Asp＋Tyr208Gln＋Asn212AspPro195Gly＋Asn198Glu＋Ser210Asp＋Leu211GlnAla194Pro＋Pro195Asn＋Asn198Asp＋Ser210AspPro195Gln＋Asn198Glu＋Val199Ser＋Ser210AspVal197His＋Asn198Glu＋Thr207Pro＋Ser210AspGly196Pro＋Val197Gly＋Asn198Asp＋Ser210GluAsn198Glu＋Ala209Gln＋Leu211Glu＋Thr214ProGln200Glu＋Tyr203Asn＋Thr207Gln＋Ala209GluPro195Ser＋Gly196Ser＋Gln200Glu＋Ala209AspPro195Ser＋Gln200Asp＋Ala209Glu＋Leu211Ala表27第6环状区-五突变的突变体Val197Ala＋Asn198Gln＋Thr207Ser＋Tyr208Ile＋Asn212AspAla194Gly＋Gly196Ser＋Val197Gly＋Gln200Glu＋Leu21lGlyGln200Asp＋Thr202Ser＋Tyr203Ser＋Pro204Ser＋Asn212SerPro195Ser＋Val199Ala＋Gly205Asn＋Ser206Glu＋Gly213SerVal199His＋Pro204Asp＋Gly205Pro＋Leu211Gly＋Thr214SerAla194Gly＋Thr207Pro＋Ser210Glu＋Leu211Cys＋Gly213GlnPro195Asn＋Gly196Pro＋Asn198Ser＋Thr202Gly＋Gly213AsnGly196Ser＋Val197Gln＋Pro204Ser＋Gly205Asp＋Leu211GlyPro195Ser＋Gly196Gln＋Pro204Asn＋Gly213Asp＋Thr214GlyPro195Gly＋Val199Asn＋Gln200Asn＋Ser210Glu＋Leu211GlyVal197Ser＋Val199Gly＋Tyr203Thr＋Gly205Ser＋Asn212GluPro195Asn＋Asn198Asp＋Thr202Ser＋Ala209His＋Asn212GlnGly196Asn＋Gly205Pro＋Thr207Pro＋Asn212Asp＋Gly213GlnVal197Gly＋Asn198Asp＋Val199His＋Gly205Gln＋Tyr208HisTyr203Val＋Thr207Gly＋Ala209Ser＋Asn212Glu＋Gly213GlnAla194Thr＋Pro195Gly＋Gly196Ser＋Tyr203Asn＋Asn212SerPro195Gln＋Thr202Gly＋Tyr203Leu＋Pro204Gly＋Ser210AspPro195Asn＋Val199Ala＋Thr202Asn＋Tyr208Pro＋Ser210AspAla194Pro＋Pro195Gly＋Asn198Glu＋Pro204Asn＋Tyr208ProAla194Ser＋Val197Thr＋Asn198Ser＋Thr202Pro＋Asn212SerGly196Ser＋Val199Pro＋Thr207Ser＋Tyr208Thr＋Asn212GlnGly205Glu＋Tyr208Gly＋Leu21lIle＋Asn212Ser＋Thr214GlyGln200Ser＋Thr202Gly＋Thr207Asp＋Leu211Ser＋Thr214GlnPro195Asn＋Thr207Asp＋Tyr208Asn＋Asn212Ser＋Gly213AsnPro195Asn＋Tyr203Ser＋Pro204Glu＋Leu211Val＋Asn212GlnGln200Asp＋Gly205Ser＋Tyr208Met＋Leu211Asn＋Asn212SerAla194Ser＋Val199Pro＋Gly205Glu＋Tyr208Met＋Gly213SerAsn198Ser＋Gln200Glu＋Thr202Gly＋Ala209Gln＋Leu211AsnGly196Pro＋Asn198Glu＋Thr202Ser＋Ala209Thr＋Thr214GlyAla194Gln＋Pro195Gly＋Gly196Asn＋Tyr203Ile＋Thr214ProPro195Gln＋Val197Cys＋Thr202Gln＋Tyr203Cys＋Tyr208HisVal199Pro＋Pro204Gly＋Gly205Ser＋Thr207Ser＋Gly213AspTyr203Gly＋Pro204Ser＋Gly205Asn＋Ala209Thr＋Ser210AspAla194Ser＋Thr207Ser＋Tyr208Cys＋Leu211Pro＋Thr214ProPro195Gly＋Val199His＋Gly205Glu＋Ala209Asn＋Leu211ProPro204Asn＋Tyr208Ile＋Ala209Gly＋Leu211Gln＋Gly213GluAla194Gln＋Val197Cys＋Asn198Ser＋Pro204Gly＋Asn212GluAsn198Glu＋Pro204Asn＋Ala209Thr＋Leu211Ser＋Gly213ProGly196Ser＋Val199Ser＋Pro204Asp＋Thr207Gly＋Tyr208IleGln200Glu＋Thr202Ser＋Tyr203Ala＋Pro204Gly＋Tyr208MetAsn198Gln＋Thr202Pro＋Gly205Asn＋Thr207Glu＋Thr214ProGly196Ser＋Val199Pro＋Ala209Gln＋Ser210Asp＋Leu211AsnAla194Gly＋Thr207Asn＋Ala209Gln＋Asn212Asp＋Thr214GlnPro195Gly＋Asn198Gln＋Val199Thr＋Tyr208His＋Leu21lIleAla194Pro＋Thr202Gly＋Thr207Ser＋Tyr208Pro＋Leu211ProGly196Ser＋Val197Ala＋Gln200Asn＋Thr202Pro＋Ser210GluVal197Gln＋Thr202Asn＋Tyr203Asn＋Gly205Asp＋Tyr208AlaGly196Pro＋Tyr203Ile＋Pro204Asn＋Leu211Ala＋Thr214GlnAla194Asn＋Gln200Asn＋Pro204Gln＋Ala209Thr＋Gly213SerAla194His＋Pro195Gly＋Gln200Ser＋Leu211Asn＋Asn212SerAla194Ser＋Gly196Ser＋Gln200Asn＋Ser206Asp＋Thr207GlyGly196Asn＋Tyr203Asn＋Tyr208Ala＋Ala209Pro＋Gly213GlnAla194Thr＋Val199Asn＋Tyr203Ser＋Tyr208His＋Asn212AspVal197Glu＋Val199Ala＋Tyr208Gln＋Ala209Gly＋Leu211ValGln200Asn＋Thr202Ser＋Tyr203Ala＋Thr207Gln＋Thr214GlyVal197Asn＋Asn198Ser＋Thr202Pro＋Thr207Gln＋Thr214GluPro195Gln＋Asn198Glu＋Gln200Ser＋Tyr203Cys＋Gly213GlnGly196Gln＋Asn198Ser＋Thr202Asn＋Tyr203Ser＋Gly205ProAla194Thr＋Thr207Pro＋Tyr208Ile＋Ala209Pro＋Thr214AsnAsn198Ser＋Val199Pro＋Thr202Asn＋Tyr203Gly＋Asn212GluAla194Asn＋Thr202Asn＋Ser210Asp＋Leu211Val＋Asn212SerVal197Ser＋Val199His＋Thr202Asn＋Ser210Asp＋Leu211CysGly196Pro＋Val197Cys＋Val199Met＋Tyr203Ser＋Leu211AsnAla194Gly＋Gly196Pro＋Val199His＋Thr202Pro＋Ser210GluGly196Ser＋Val197His＋Asn198Glu＋Thr202Pro＋Asn212SerAla194His＋Gly196Asn＋Val197Gln＋Thr202Ser＋Gly205GluPro195Gly＋Gly196Pro＋Asn198Ser＋Gln200Ser＋Thr202ProVal197Asn＋Asn198Asp＋Gln200Asn＋Thr202Gln＋Tyr208AlaPro195Asn＋Thr202Ser＋Pro204Asp＋Tyr208Ala＋Ala209SerTyr203His＋Pro204Asn＋Gly205Asp＋Thr207Gln＋Leu211GlyAla194Asn＋Pro195Gln＋Gly196Gln＋Pro204Asn＋Gly213GluVal197Thr＋Val199His＋Ser206Glu＋Gly213Asn＋Thr214AsnGly196Gln＋Val197Asn＋Tyr203Thr＋Pro204Ser＋Thr214AspGly196Asn＋Val199Gly＋Gln200Ser＋Tyr203Asn＋Gly213GluAla194Thr＋Gln200Asn＋Thr202Ser＋Gly205Gln＋Gly213AspPro195Ser＋Asn198Ser＋Val199Gly＋Tyr203Val＋Thr207GluAla194Asn＋Pro195Gly＋Thr202Gly＋Tyr203Gly＋Leu211AspPro195Ser＋Val197Ser＋Tyr203Cys＋Gly205Glu＋Ala209ThrGly196Gln＋Val197Gly＋Gly205Ser＋Tyr208Leu＋Thr214AspAla194Gln＋Gly196Asn＋Tyr203Pro＋Pro204Ser＋Gly213AspGly196Gln＋Thr202Gly＋Tyr203Cys＋Pro204Gln＋Ser206GluVal197Pro＋Val199Met＋Tyr203Val＋Pro204Gly＋Thr214AspGly196Pro＋Gln200Asn＋Tyr203Cys＋Ala209Thr＋Thr214AsnAsn198Gln＋Pro204Asn＋Thr207Gln＋Gly213Pro＋Thr214AspPro195Gly＋Val197Ser＋Val199Ala＋Thr207Asp＋Leu211GlyPro195Gln＋Asn198Gln＋Ser206Asp＋Thr207Ser＋Ala209SerAla194Ser＋Tyr203Leu＋Ala209Asn＋Ser210Asp＋Thr214SerVal199His＋Thr202Ser＋Gly205Asn＋Leu211Pro＋Asn212GlnGly196Ser＋Asn198Ser＋Thr207Asn＋Tyr208Gln＋Ser210GluPro195Asn＋Asn198Gln＋Thr207Ser＋Gly213Asn＋Thr214GlyGly196Gln＋Gln200Ser＋Pro204Asn＋Gly205Gln＋Gly213AspPro204Asn＋Ser206Glu＋Tyr208Leu＋Ala209Gly＋Asn212GlnVal197Thr＋Val199Gly＋Gln200Asp＋Thr202Pro＋Thr207GlyPro195Gln＋Val199Ser＋Thr202Asn＋Gly205Asp＋Ala209GlnGly196Gln＋Tyr203Thr＋Tyr208Cys＋Leu211Ala＋Thr214GluAsn198Gln＋Gln200Ser＋Pro204Gln＋Asn212Ser＋Gly213ProVal197Pro＋Val199Asn＋Thr202Gln＋Tyr203Asn＋Gly213SerAla194Ser＋Gln200Ser＋Thr202Gln＋Thr207Gln＋Ala209GluThr202Gly＋Tyr203Thr＋Tyr208Pro＋Gly213Asp＋Thr214GluVal197Gln＋Pro204Gly＋Gly205Ser＋Ala209Glu＋Ser210AspAla194Ser＋Val199Ala＋Tyr208Ala＋Ala209Glu＋Ser210GluVal199Ala＋Gln200Ser＋Gly205Pro＋Asn212Glu＋Gly213AspAla194Pro＋Thr202Ser＋Pro204Glu＋Gly205Asp＋Ala209ProPro204Glu＋Gly205Glu＋Thr207Gly＋Tyr208Ser＋Leu211IlePro195Gln＋Val197Thr＋Pro204Glu＋Gly205Asp＋Leu211ValVal199His＋Gly205Asp＋Ser206Glu＋Ala209Gln＋Asn212GlnTyr203Ala＋Gly205Glu＋Ser206Asp＋Tyr208Ile＋Asn212GlnPro195Asn＋Gly205Glu＋Ser206Asp＋Tyr208Ser＋Ala209SerVal197His＋Pro204Gly＋Gly205Glu＋Ser206Asp＋Tyr208CysThr202Asn＋Pro204Ser＋Gly205Asp＋Ser206Asp＋Tyr208ThrAla194Asn＋Thr202Ser＋Tyr203Gly＋Ser206Glu＋Thr207GluPro195Ser＋Gly196Gln＋Val199Thr＋Ser206Glu＋Thr207AspGly196Pro＋Ser206Asp＋Thr207Glu＋Tyr208Thr＋Leu211HisAla194His＋Gly196Gln＋Pro204Asn＋Tyr208Asp＋Ala209AspVal199Ala＋Tyr203Pro＋Tyr208Asp＋Ala209Asp＋Asn212GlnVal199Ala＋Thr202Ser＋Thr207Glu＋Tyr208Asp＋Thr214SerGly196Asn＋Thr202Gln＋Pro204Ser＋Thr207Asp＋Tyr208AspPro195Gln＋Gln200Glu＋Thr202Pro＋Tyr203Pro＋Ser210AspGly196Pro＋Val197Thr＋Asn198Gln＋Gln200Asp＋Ser210AspGly196Gln＋Asn198Glu＋Thr207Pro＋Tyr208Ser＋Ser210GluAla194Pro＋Asn198Glu＋Thr207Pro＋Ala209His＋Ser210GluAsn198Glu＋Val199Gln＋Pro204Ser＋Tyr208Asn＋Ser210AspPro195Gly＋Asn198Glu＋Thr207Gly＋Tyr208Cys＋Ser210AspAsn198Asp＋Val199Thr＋Leu211Asp＋Asn212Ser＋Thr214GlyGly196Pro＋Asn198Gln＋Gln200Glu＋Tyr203Asn＋Ala209GluGly196Pro＋Tyr203Thr＋Pro204Asp＋Ser206Asp＋Ala209GlnPro195Asn＋Gly196Pro＋Asn198Asp＋Ala209Pro＋Asn212GluPro195Asn＋Gly196Gln＋Gly205Asp＋Thr207Glu＋Leu211ThrThr202Asn＋Pro204Asp＋Thr207Asp＋Tyr208Met＋Thr214ProTyr203Gly＋Pro204Asp＋Thr207Asp＋Asn212Gln＋Thr214GlyPro195Ser＋Tyr203His＋Pro204Glu＋Thr207Asp＋Gly213GlnAsn198Glu＋Gln200Glu＋Tyr203Cys＋Pro204Gly＋Asn212GlnVal197Glu＋Val199Gly＋Thr202Gly＋Tyr203Ser＋Gly213GluPro204Gly＋Thr207Asn＋Ala209His＋Ser210Asp＋Asn212AspPro195Gly＋Gly196Gln＋Thr202Gln＋Ser210Asp＋Asn212AspThr202Asn＋Tyr203Asn＋Gly205Pro＋Ser210Asp＋Asn212AspThr202Gln＋Tyr203Gln＋Ser210Asp＋Asn212Asp＋Gly213GlnGly196Ser＋Ser210Asp＋Leu211Gly＋Asn212Asp＋Thr214GlnGly196Pro＋Ala209Ser＋Ser210Glu＋Leu211Asn＋Asn212AspGly205Ser＋Ala209Pro＋Ser210Glu＋Asn212Asp＋Thr214ProPro195Gly＋Tyr203Gly＋Ser210Asp＋Asn212Asp＋Thr214SerPro195Gln＋Val197Asn＋Val199Cys＋Tyr208Glu＋Ser210AspVal197Ser＋Tyr203Gly＋Pro204Glu＋Thr207Ser＋Tyr208AspPro195Ser＋Gly196Asn＋Val197Asp＋Val199Gly＋Ser210GluGly196Pro＋Val197Asp＋Thr202Pro＋Ser210Glu＋Thr214ProVal197Glu＋Val199His＋Tyr208Thr＋Leu211Gln＋Thr214GluAsn198Asp＋Val199Asn＋Thr207Asn＋Ala209His＋Gly213AspVal197His＋Asn198Glu＋Tyr203Gln＋Tyr208Cys＋Gly213AspPro195Asn＋Val197Asn＋Asn198Glu＋Thr202Pro＋Gly213AspPro195Ser＋Asn198Glu＋Tyr203Ser＋Gly205Ser＋Ala209AspPro195Gly＋Gly196Asn＋Asn198Asp＋Val199Met＋Ala209GluPro195Gln＋Asn198Glu＋Thr202Asn＋Pro204Gly＋Ala209AspVal197Glu＋Gln200Glu＋Thr202Gly＋Tyr208Ala＋Gly213GlnVal197Asp＋Gln200Asp＋Thr207Ser＋Tyr208Ile＋Ala209SerVal197Asp＋Gln200Glu＋Ala209Ser＋Leu211Gly＋Asn212SerVal197Glu＋Gln200Asp＋Thr202Asn＋Gly205Gln＋Asn212GlnAla194Pro＋Gly196Pro＋Val197Met＋Gln200Glu＋Asn212AspGly196Gln＋Gln200Glu＋Thr202Pro＋Tyr203Ile＋Asn212GluAla194His＋Gln200Asp＋Asn212Glu＋Gly213Pro＋Thr214AsnVal199Pro＋Gln200Glu＋Pro204Gly＋Asn212Glu＋Gly213SerAsn198Ser＋Gln200Asp＋Gly205Pro＋Thr207Ser＋Asn212GluAla194Asn＋Gln200Asp＋Gly205Asn＋Tyr208Asn＋Asn212AspPro195Asn＋Gln200Ser＋Thr202Pro＋Pro204Asp＋Ala209GluPro195Ser＋Thr202Asn＋Tyr203Cys＋Pro204Glu＋Ala209AspAla194His＋Asn198Ser＋Thr202Asn＋Pro204Glu＋Ala209AspAla194His＋Pro204Asn＋Ser206Glu＋Ala209Glu＋Thr214SerPro195Gly＋Val197Cys＋Ser206Glu＋Ala209Glu＋Asn212SerThr202Gly＋Ser206Glu＋Thr207Ser＋Tyr208Met＋Ala209GluAla194Gly＋Gln200Glu＋Thr207Asp＋Leu211Ser＋Thr214ProPro195Asn＋Val199Ser＋Gln200Glu＋Gly205Asn＋Thr207AspAla194Gln＋Asn198Gln＋Gly205Pro＋Ser210Asp＋Gly213AspGly196Ser＋Thr207Gly＋Ser210Glu＋Leu211Thr＋Gly213AspAsn198Gln＋Pro204Gln＋Thr207Asp＋Tyr208Thr＋Ser210GluAla194Ser＋Gly196Gln＋Thr207Glu＋Ser210Glu＋Leu211ThrThr207Glu＋Ser210Glu＋Asn212Ser＋Gly213Ser＋Thr214SerVal197Cys＋Thr207Glu＋Ala209His＋Ser210Glu＋Asn212SerAsn198Glu＋Val199Met＋Tyr208Ala＋Gly213Gln＋Thr214AspAla194Gln＋Asn198Glu＋Ala209Pro＋Leu21lHis＋Thr214AspAsn198Asp＋Thr207Pro＋Leu211Gly＋Asn212Ser＋Thr214AspAla194Ser＋Gly196Asn＋Asn198Asp＋Pro204Gln＋Thr214Glu表28第6环状区-六突变的突变体Ala194Gln＋Val197Ser＋Thr202Ser＋Tyr203Ser＋Ala209Ser＋Asn212AspPro195Ser＋Val199Cys＋Gly205Ser＋Leu211Thr＋Asn212Glu＋Gly213ProVal197Thr＋Asn198Ser＋Val199Pro＋Gln200Ser＋Thr202Ser＋Leu211AsnThr202Gly＋Pro204Ser＋Gly205Asn＋Tyr208Asp＋Gly213Gln＋Thr214ProVal197Gly＋Gln200Glu＋Thr202Gly＋Tyr208Ala＋Leu211Val＋Asn212SerAla194Pro＋Gly196Ser＋Val199Cys＋Tyr203Ala＋Pro204Ser＋Ala209AspPro195Asn＋Val199Pro＋Thr202Asn＋Tyr203Gly＋Thr207Asp＋Gly213AsnAla194Gln＋Thr202Ser＋Tyr203Ile＋Pro204Asn＋Gly205Asp＋Tyr208SerAla194Gln＋Val197His＋Ala209Ser＋Ser210Asp＋Leu211Gly＋Asn212GlnThr202Gly＋Gly205Pro＋Ser210Asp＋Leu211Ala＋Gly213Ser＋Thr214GlnAla194His＋Gln200Ser＋Thr202Gly＋Pro204Ser＋Tyr208His＋Gly213GluAla194Asn＋Pro195Gln＋Gly205Asn＋Tyr208Ser＋Leu211Val＋Thr214ProGly196Pro＋Asn198Ser＋Gln200Ser＋Thr202Asn＋Tyr208Ile＋Thr214GluPro195Asn＋Gly205Pro＋Thr207Gln＋Ser210Glu＋Leu211Asn＋Gly213ProAla194Ser＋Val197Gly＋Asn198Asp＋Tyr203Cys＋Gly205Gln＋Leu211MetAla194Ser＋Val197Asp＋Val199Gln＋Tyr203Cys＋Gly205Ser＋Gly213GlnAla194Pro＋Thr202Gly＋Thr207Ser＋Tyr208Met＋Leu211Pro＋Gly213GluAla194Thr＋Pro195Asn＋Gly196Gln＋Gln200Ser＋Thr202Asn＋Pro204GluAla194Gln＋Gly196Asn＋Val197Ser＋Asn198Ser＋Val199Thr＋Ser210GluGly196Ser＋Val199Gly＋Tyr203Ile＋Pro204Asn＋Leu211Asp＋Thr214GlnAla194Asn＋Gln200Asn＋Pro204Gln＋Ala209Thr＋Leu211Asn＋Gly213SerAla194His＋Pro195Gly＋Gly196Asn＋Gln200Ser＋Tyr203Ser＋Asn212SerAla194Pro＋Val197Met＋Val199Asn＋Thr202Ser＋Ser206Asp＋Thr207AsnPro195Gln＋Thr202Pro＋Gly205Asn＋Ala209Pro＋Asn212Ser＋Thr214AsnPro195Asn＋Asn198Glu＋Val199His＋Pro204Asn＋Thr207Gly＋Leu211MetGly196Ser＋Pro204Asn＋Thr207Pro＋Asn212Asp＋Gly213Ser＋Thr214ProAla194Ser＋Pro195Ser＋Val197Cys＋Pro204Asn＋Thr207Glu＋Gly213GlnAla194His＋Pro195Gly＋Val197Gly＋Val199Gln＋Thr207Gly＋Asn212GluAla194Gln＋Pro1955er＋Val197Ser＋Tyr203Met＋Leu211Glu＋Gly213AsnAla194Asn＋Gly196Asn＋Pro204Asp＋Thr207Pro＋Tyr208Ser＋Thr214SerPro195Asn＋Thr202Gly＋Gly205Asp＋Thr207Gln＋Tyr208Thr＋Ala209AsnAla194His＋Pro195Gly＋Val197His＋Gly205Gln＋Ala209Glu＋Thr214AsnAla194Gln＋Asn198Gln＋Tyr203Met＋Ala209Gln＋Ser210Glu＋Leu211GlyAla194Pro＋Gly196Ser＋Gln200Ser＋Thr202Pro＋Thr207Asp＋Ala209GlyPro195Gln＋Gly196Ser＋Asn198Gln＋Gly205Ser＋Ser210Asp＋Leu211ValAsn198Gln＋Val199Met＋Gln200Glu＋Thr202Gly＋Gly205Asn＋Tyr208AlaAla194Pro＋Val199Ala＋Pro204Asn＋Thr207Gln＋Leu211Glu＋Gly213SerGly196Pro＋Val197Thr＋Val199His＋Gly205Asn＋Ser206Asp＋Thr207AsnPro195Asn＋Gly196Ser＋Thr202Asn＋Tyr203Gly＋Leu211Asn＋Asn212GluAla194Ser＋Pro195Gln＋Val197Cys＋Ser206Glu＋Tyr208His＋Asn212GlnPro195Gln＋Gly196Pro＋Val197Thr＋Ser210Asp＋Gly213Ser＋Thr214ProAla194Gly＋Pro195Ser＋Thr202Gly＋Pro204Glu＋Tyr208Thr＋Gly213ProAla194Asn＋Pro195Asn＋Gly196Asn＋Gly205Pro＋Ala209Asn＋Ser210AspPro195Ser＋Val199Gln＋Tyr208Asn＋Ala209Gln＋Leu211Gly＋Gly213GlnAsn198Asp＋Val199Ala＋Thr202Asn＋Ala209Pro＋Asn212Gln＋Thr214GlyPro195Asn＋Gly196Gln＋Val199Pro＋Ser206Glu＋Tyr208Ala＋Leu211ValAla194Gln＋Pro195Ser＋Asn198Asp＋Val199Ser＋Thr202Pro＋Ala209GlnPro195Gly＋Val199Met＋Gly205Glu＋Thr207Asn＋Ala209Pro＋Gly213ProPro195Asn＋Gly196Gln＋Gln200Asp＋Thr207Asn＋Ala209Gly＋Asn212SerGly196Gln＋Val197His＋Thr207Asn＋Ser210Asp＋Leu211Ala＋Gly213GlnAla194Gly＋Val197Thr＋Tyr203Met＋Gly205Glu＋Ala209Gly＋Thr214GlyGln200Glu＋Thr202Asn＋Tyr203Gly＋Thr207Asn＋Asn212Gln＋Gly213ProGly196Asn＋Thr202Ser＋Pro204Glu＋Leu211Pro＋Gly213Ser＋Thr214AsnPro195Gly＋Val197Asn＋Val199Gln＋Gln200Asn＋Thr207Gly＋Asn212SerPro195Gln＋Gly196Pro＋Val197His＋Tyr203Gly＋Ser206Asp＋Thr214SerGly196Gln＋Tyr203His＋Thr207Asn＋Leu211Asn＋Gly213Pro＋Thr214AspAla194Asn＋Asn198Ser＋Gln200Asn＋Thr202Ser＋Tyr203Ile＋Ala209ThrAla194His＋Val197Cys＋Gln200Ser＋Tyr203Gly＋Thr207Gly＋Ala209HisAsn198Gln＋Val199Thr＋Gln200Asp＋Pro204Asn＋Asn212Ser＋Gly213GlnGly196Pro＋Tyr203Gln＋Pro204Ser＋Thr207Asn＋Tyr208Met＋Leu211AspGly196Ser＋Val197Met＋Asn198Ser＋Thr202Asn＋Thr207Asp＋Asn212GlnPro195Gly＋Gly196Pro＋Thr202Asn＋Tyr203Ile＋Gly213Asp＋Thr214AsnPro195Gly＋Gly196Pro＋Gln200Glu＋Pro204Ser＋Tyr208Ile＋Gly213SerVal199Ala＋Thr202Gln＋Gly205Ser＋Ala209Asn＋Ser210Glu＋Leu211CysAla194His＋Pro195Ser＋Val199Thr＋Gln200Glu＋Tyr203Pro＋Tyr208CysGly196Gln＋Pro204Gly＋Ser206Asp＋Thr207Pro＋Ala209Gln＋Gly213SerPro195Asn＋Thr202Asn＋Pro204Asn＋Ala209Gln＋Asn212Asp＋Gly213GlnGly196Ser＋Asn198Gln＋Tyr203His＋Asn212Ser＋Gly213Asp＋Thr214GlyAla194Gln＋Val197Pro＋Val199Gln＋Gln200Asn＋Tyr208Glu＋Thr214SerVal199Cys＋Gln200Asn＋Pro204Gly＋Ser206Asp＋Thr207Ser＋Asn212SerAla194Gln＋Gly196Gln＋Val199His＋Thr202Pro＋Tyr203Thr＋Gly205SerAsn198Gln＋Val199Gly＋Thr202Ser＋Ser210Asp＋Asn212Ser＋Gly213GlnVal199Thr＋Gln200Asn＋Pro204Ser＋Tyr208Val＋Asn212Gln＋Thr214AspAla194His＋Gly196Gln＋Val199Gln＋Thr202Gly＋Pro204Asp＋Leu211CysAla194Ser＋Val197Gly＋Pro204Gln＋Ala209Pro＋Ser210Glu＋Thr214GlyAla194Gln＋Gly196Pro＋Tyr203Gln＋Thr207Pro＋Tyr208Pro＋Ala209AsnGly196Ser＋Thr202Asn＋Tyr208Ile＋Ala209Gln＋Leu211Ala＋Gly213GluPro195Asn＋Gly196Pro＋Val197Glu＋Gln200Asn＋Tyr203Ser＋Tyr208SerGly196Ser＋Val199His＋Gly205Asp＋Tyr208Val＋Ala209Gly＋Thr214ProVal197Ser＋Val199Met＋Gln200Asn＋Thr202Ser＋Tyr208Cys＋Gly213GlnAla194Ser＋Gly196Gln＋Val199Pro＋Leu211Val＋Gly213Glu＋Thr214GlyAla194Gln＋Asn198Gln＋Val199Cys＋Thr207Gly＋Leu211Asp＋Asn212GluAla194Thr＋Asn198Gln＋Thr202Asn＋Pro204Ser＋Gly213Glu＋Thr214AspAla194Thr＋Pro195Ser＋Val199Ala＋Ala209Glu＋Ser210Glu＋Leu211CysTyr203Leu＋Gly205Ser＋Ala209Glu＋Ser210Glu＋Asn212Ser＋Gly213AsnVal197Glu＋Asn198Glu＋Tyr203Pro＋Thr207Gly＋Ala209Ser＋Asn212SerVal197Asp＋Asn198Asp＋Val199His＋Thr202Ser＋Pro204Gln＋Ala209AsnGly196Asn＋Val197Glu＋Asn198Asp＋Gln200Ser＋Pro204Asn＋Thr207GlyPro195Gln＋Thr202Ser＋Tyr203Ser＋Thr207Gln＋Asn212Asp＋Gly213GluGly196Gln＋Tyr203His＋Gly205Ser＋Leu211Asn＋Asn212Asp＋Gly213GluAsn198Ser＋Gly205Ser＋Tyr208Ile＋Asn212Glu＋Gly213Glu＋Thr214ProAla194Ser＋Gly196Gln＋Pro204Glu＋Gly205Asp＋Tyr208Asn＋Thr214AsnAla194Ser＋Pro195Gly＋Val199Cys＋Pro204Asp＋Gly205Glu＋Asn212GlnAla194Thr＋Val199Pro＋Gly205Asn＋Ala209Gln＋Ser210Glu＋Leu211AspThr202Gln＋Thr207Gly＋Ser210Asp＋Leu211Glu＋Gly213Gln＋Thr214SerThr202Gln＋Gly205Asp＋Ser206Glu＋Tyr208Met＋Gly213Gln＋Thr214AsnAla194Gly＋Gly205Glu＋Ser206Glu＋Ala209Gly＋Leu211Gln＋Thr214GlyGln200Ser＋Thr202Ser＋Tyr203Gly＋Gly205Asp＋Ser206Glu＋Thr214GlnVal197Asn＋Thr202Ser＋Gly205Glu＋Ser206Glu＋Ala209His＋Thr214SerAla194Gln＋Thr202Ser＋Tyr203Gly＋Ser206Asp＋Thr207Glu＋Gly213SerGly196Pro＋Val199Gln＋Tyr203Met＋Ser206Glu＋Thr207Asp＋Ala209ThrPro195Gly＋Gly196Pro＋Thr202Ser＋Pro204Gln＋Tyr208Asp＋Ala209GluPro195Gln＋Gly196Pro＋Val197Pro＋Gln200Asp＋Gly205Ser＋Ser210AspGly196Ser＋Asn198Glu＋Gly205Pro＋Ala209His＋Ser210Glu＋Asn212GlnAla194Thr＋Asn198Asp＋Thr202Ser＋Tyr203Cys＋Ala209Pro＋Ser210GluGly196Pro＋Asn198Glu＋Gly205Ser＋Ser210Glu＋Leu211Asn＋Thr214AsnAla194Pro＋Gly196Ser＋Asn198Glu＋Gly205Ser＋Thr207Pro＋Ser210GluGly196Ser＋Val197His＋Asn198Asp＋Gln200Ser＋Thr207Gln＋Ser210AspAsn198Glu＋Val199Met＋Gly205Pro＋Ala209His＋Ser210Glu＋Thr214ProVal197Ser＋Asn198Glu＋Pro204Ser＋Thr207Ser＋Ala209Pro＋Leu21lAspPro195Ser＋Gln200Asp＋Thr202Asn＋Tyr203Ser＋Ala209Asp＋Asn212SerGln200Asp＋Pro204Ser＋Gly205Pro＋Tyr208His＋Ala209Asp＋Leu211AsnPro204Glu＋Ser206Asp＋Tyr208Ile＋Leu211Gly＋Asn212Gln＋Thr214AsnGly196Ser＋Asn198Ser＋Val199Ala＋Pro204Asp＋Ser206Glu＋Tyr208SerPro195Asn＋Pro204Asp＋Ser206Asp＋Tyr208Ser＋Leu211Ser＋Asn212GlnAla194Gly＋Asn198Glu＋Gln200Ser＋Thr207Gly＋Ala209Asn＋Asn212GluAsn198Glu＋Gly205Gln＋Ala209Pro＋Leu211Cys＋Asn212Asp＋Gly213ProAla194Thr＋Val197Thr＋Asn198Gln＋Pro204Gly＋Gly205Asp＋Thr207AspAla194Asn＋Gly196Pro＋Pro204Asp＋Thr207Asp＋Tyr208Cys＋Gly213AsnAla194Thr＋Pro195Gln＋Gln200Ser＋Pro204Glu＋Thr207Asp＋Gly213ProVal197Pro＋Asn198Glu＋Gln200Asp＋Len211Ala＋Asn212Ser＋Gly213ProAsn198Asp＋Gln200Glu＋Tyr203Val＋Leu211Thr＋Asn212Ser＋Gly213ProPro195Gln＋Asn198Asp＋Val199Gln＋Gln200Glu＋Pro204Asn＋Ala209ProAla194Gly＋Gly196Asn＋Asn198Asp＋Gln200Asp＋Thr207Gly＋Asn212SerGln200Asp＋Thr202Asn＋Tyr203Ala＋Thr207Gln＋Leu211Glu＋Thr214ProVal197Asp＋Val199His＋Tyr203Val＋Tyr208His＋Leu211Asp＋Gly213ProPro195Asn＋Val197Gln＋Val199Gln＋Thr207Asp＋Ala209Glu＋Gly213SerAsn198Gln＋Gln200Asn＋Tyr203Ala＋Thr207Asp＋Tyr208Val＋Ala209AspVal197Ala＋Asn198Gln＋Pro204Ser＋Tyr208Thr＋Ser210Glu＋Asn212GluGly196Asn＋Gln200Asn＋Pro204Gln＋Ser210Asp＋Asn212Glu＋Thr214ProPro195Gln＋Val199Pro＋Ser210Asp＋Asn212Glu＋Gly213Asn＋Thr214GlyPro195Gly＋Gln200Ser＋Thr207Pro＋Tyr208Gln＋Ser210Glu＋Asn212GluPro195Gly＋Val199Cys＋Thr202Asn＋Tyr208His＋Ser210Glu＋Asn212AspVal199His＋Thr202Gly＋Pro204Gln＋Ala209Thr＋Ser210Asp＋Asn212AspAsn198Gln＋Thr202Pro＋Ala209Ser＋Ser210Glu＋Asn212Glu＋Gly213ProAla194Ser＋Val199Gln＋Tyr208Gly＋Ser210Asp＋Leu211Gln＋Asn212AspAla194Asn＋Val199Gly＋Gly205Ser＋Ala209His＋Ser210Glu＋Asn212GluAla194Ser＋Thr202Asn＋Pro204Gly＋Tyr208Glu＋Ser210Glu＋Gly213GlnPro195Gln＋Val199Asn＋Thr207Gly＋Tyr208Asp＋Ser210Asp＋Leu211IlePro195Gln＋Gln200Asn＋Tyr203Gln＋Tyr208Glu＋Ser210Glu＋Asn212GlnAla194His＋Tyr203Asn＋Tyr208Asp＋Ser210Asp＋Leu211His＋Thr214SerGly196Asn＋Val197Gln＋Thr202Asn＋Tyr208Glu＋Ala209Asn＋Ser210GluAla194Asn＋Gln200Ser＋Tyr203Met＋Ser206Glu＋Tyr208Glu＋Leu211ValAsn198Ser＋Pro204Gly＋Ser206Glu＋Tyr208Asp＋Asn212Gln＋Thr214GlyAla194Pro＋Gln200Asn＋Thr202Pro＋Tyr203Met＋Asn212Asp＋Thr214AspPro195Ser＋Asn198Glu＋Gln200Ser＋Thr202Asn＋Gly205Pro＋Gly213AspPro195Ser＋Asn198Glu＋Val199Ala＋Ala209Glu＋Leu211Asn＋Thr214GlyPro195Gly＋Asn198Glu＋Gln200Asn＋Thr202Pro＋Tyr203Ile＋Ala209GluVal197Ser＋Asn198Asp＋Tyr203Pro＋Tyr208Met＋Ala209Asp＋Leu211GlnPro195Ser＋Asn198Asp＋Gln200Ser＋Tyr208Met＋Ala209Asp＋Leu211MetGly196Asn＋Val197Asp＋Val199Gln＋Gln200Glu＋Gly205Asn＋Gly213ProGln200Glu＋Thr202Gln＋Tyr203Met＋Ala209Asn＋Leu211Ala＋Asn212AspAla194Pro＋Gly196Pro＋Gln200Asp＋Thr202Asn＋Tyr208Ser＋Asn212GluVal197Gly＋Val199Ser＋Gln200Glu＋Tyr203Gly＋Asn212Glu＋Gly213ProPro204Asp＋Thr207Gln＋Tyr208Leu＋Ala209Asp＋Asn212Ser＋Thr214AsnAla194Ser＋Pro195Gln＋Thr202Ser＋Pro204Asp＋Ala209Asp＋Thr214AsnPro195Asn＋Thr202Ser＋Tyr203Leu＋Tyr208Asp＋Ala209His＋Leu211AspAla194Asn＋Gly196Pro＋Ser206Glu＋Tyr208His＋Ala209Asp＋Thr214GlyPro204Gln＋Gly205Asn＋Ser206Glu＋Thr207Gly＋Ala209Asp＋Leu211CysVal197Gln＋Thr202Gln＋Pro204Gly＋Ser206Asp＋Ala209Asp＋Thr214SerAla194Gly＋Val197Gln＋Asn198Gln＋Tyr203Val＋Ser206Asp＋Ala209AspGly196Ser＋Val197Asn＋Gln200Asp＋Thr202Pro＋Thr207Asp＋Asn212GlnAla194Gly＋Gly196Ser＋Ser210Asp＋Leu211Ser＋Asn212Gln＋Gly213GluVal197Thr＋Tyr203His＋Thr207Gln＋Tyr208Pro＋Ser210Asp＋Gly213AspAsn198Ser＋Val199Cys＋Tyr203Val＋Gly205Ser＋Ser210Asp＋Gly213AspVal197His＋Val199Asn＋Pro204Gln＋Ser210Asp＋Asn212Ser＋Gly213AspGly196Ser＋Val197Ala＋Gln200Asn＋Thr202Pro＋Ser210Glu＋Gly213GluAsn198Glu＋Tyr203Gly＋Gly205Gln＋Thr207Pro＋Tyr208Val＋Thr214GluAla194Gly＋Gly196Asn＋Asn198Asp＋Tyr203Leu＋Tyr208His＋Thr214GluPro195Gly＋Asn198Glu＋Val199Cys＋Ala209Gln＋Asn212Gln＋Thr214AspAsn198Asp＋Val199Asn＋Thr202Asn＋Pro204Gln＋Asn212Gln＋Thr214AspVal199Pro＋Gln200Asp＋Tyr203Asn＋Leu211Pro＋Gly213Glu＋Thr214AsnPro195Ser＋Gly196Pro＋Val199Ala＋Gln200Asp＋Asn212Gln＋Gly213AspAla194Thr＋Pro195Gly＋Gly196Pro＋Gln200Glu＋Gly205Asn＋Gly213GluVal199His＋Gln200Glu＋Pro204Ser＋Leu211His＋Gly213Glu＋Thr214GlnAsn198Asp＋Gln200Ser＋Gly205Ser＋Tyr208Asp＋Leu211Ala＋Asn212SerAla194Ser＋Asn198Asp＋Thr202Ser＋Tyr208Asp＋Ala209Pro＋Gly213GlnGln200Asn＋Tyr203Pro＋Pro204Glu＋Gly205Pro＋Ser210Asp＋Leu211ProVal197Ser＋Tyr203Pro＋Pro204Asp＋Ala209Gly＋Ser210Asp＋Thr214AsnGly196Asn＋Pro204Glu＋Ser210Glu＋Leu211Pro＋Asn212Ser＋Thr214Gly表29第6环状区-七突变的突变体Pro195Ser＋Asn198Ser＋Val199Met＋Ser206Glu＋Thr207Ser＋Gly213Asn＋Thr214GlnAla194Gln＋Pro195Asn＋Val197Ser＋Thr202Ser＋Tyr203Ser＋Ala209Ser＋Asn212AspVal197Asp＋Gln200Asn＋Thr202Pro＋Gly205Gln＋Thr207Gly＋Tyr208Met＋Leu211ValAla194Ser＋Pro195Gly＋Asn198Ser＋Thr202Pro＋Ser206Glu＋Tyr208Ala＋Thr214GlnAla194Pro＋Pro195Asn＋Thr202Gly＋Tyr203Thr＋Ser206Glu＋Leu211Ser＋Thr214ProGly196Gln＋Val197Thr＋Gln200Asn＋Tyr203Cys＋Gly205Ser＋Ser210Glu＋Leu211ValAla194His＋Val199Ser＋Thr202Gly＋Thr207Asn＋Leu211Met＋Gly213Glu＋Thr214ProVal199Gly＋Tyr203Cys＋Pro204Gly＋Thr207Asn＋Ala209His＋Asn212Asp＋Gly213AsnAla194Gln＋Gly196Pro＋Tyr203Ser＋Pro204Ser＋Tyr208Ser＋Leu211Glu＋Thr214ProGly196Gln＋Gly205Asn＋Thr207Asp＋Tyr208Met＋Asn212Gln＋Gly213Ser＋Thr214ProGly196Gln＋Val199Cys＋Ser206Glu＋Thr207Pro＋Ala209Pro＋Leu211Ile＋Gly213SerThr202Pro＋Tyr203Val＋Pro204Asn＋Ser206Asp＋Asn212Ser＋Gly213Ser＋Thr214GlyAla194Thr＋Pro195Asn＋Gly196Pro＋Gln200Asn＋Pro204Ser＋Asn212Glu＋Gly213GlnAsn198Gln＋Gln200Ser＋Pro204Gln＋Gly205Asn＋Thr207Glu＋Asn212Ser＋Gly213SerAla194Ser＋Gly196Ser＋Asn198Ser＋Tyr203Gly＋Gly205Pro＋Ser206Glu＋Gly213ProAla194Thr＋Gly196Gln＋Val197Pro＋Thr202Gln＋Pro204Ser＋Ala209Gly＋Asn212GluPro195Gln＋Gly196Pro＋Asn198Gln＋Pro204Ser＋Gly205Glu＋Tyr208Gln＋Gly213AsnVal199Thr＋Tyr203Gln＋Pro204Gln＋Gly205Asn＋Thr207Glu＋Leu211Thr＋Thr214GlnVal197Thr＋Asn198Glu＋Thr202Gly＋Gly205Pro＋Thr207Pro＋Tyr208Met＋Gly213GlnPro195Gly＋Gly196Ser＋Thr202Gly＋Tyr203Gln＋Pro204Gln＋Tyr208Pro＋Thr214GlnPro195Asn＋Asn198Asp＋Gln200Ser＋Pro204Gly＋Gly205Gln＋Tyr208Gly＋Ala209AsnPro195Asn＋Val199His＋Pro204Asn＋Ala209Pro＋Leu211Thr＋Asn212Ser＋Thr214ProAsn198Gln＋Val199Thr＋Gln200Asp＋Pro204Asn＋Leu211Val＋Asn212Ser＋Gly213GlnPro195Ser＋Gly196Ser＋Val197Met＋Val199Gly＋Gly205Pro＋Ser210Glu＋Asn212GlnAla194Thr＋Pro195Gln＋Asn198Gln＋Thr202Asn＋Thr207Asp＋Tyr208Thr＋Gly213SerPro195Gln＋Gly196Asn＋Gly205Glu＋Thr207Pro＋Tyr208Asn＋Ala209His＋Asn212SerAsn198Gln＋Thr202Pro＋Thr207Pro＋Leu211Pro＋Asn212Glu＋Gly213Asn＋Thr214SerAla194Pro＋Val197Met＋Pro204Ser＋Thr207Gln＋Tyr208Asn＋Ala209Gln＋Thr214GlyAla194Asn＋Val197Cys＋Pro204Asn＋Tyr208Thr＋Ala209Pro＋Ser210Glu＋Leu211SerGly196Asn＋Val197Gly＋Tyr203Pro＋Gly205Asp＋Leu211Ala＋Gly213Asn＋Thr214SerAla194Thr＋Pro195Asn＋Tyr203Pro＋Gly205Ser＋Ser210Glu＋Leu211Ala＋Gly213ProAla194His＋Gly196Asn＋Val197Met＋Asn198Ser＋Ser206Asp＋Leu211Asn＋Asn212SerAla194Thr＋Gly196Ser＋Val199Ser＋Thr202Asn＋Ala209Pro＋Leu211Pro＋Gly213GlnGly196Gln＋Val199His＋Thr202Pro＋Tyr203Thr＋Pro204Asp＋Gly205Ser＋Ala209AsnVal197Cys＋Val199Gly＋Thr202Gln＋Tyr203His＋Gly205Glu＋Tyr208Ile＋Thr214GlnAla194Gly＋Val197Cys＋Asn198Asp＋Gln200Ser＋Tyr203Leu＋Ala209Gln＋Asn212GlnAla194Gly＋Tyr203Cys＋Gly205Gln＋Ser206Glu＋Tyr208Met＋Leu211Cys＋Thr214GlyGly196Ser＋Val199Gln＋Thr202Ser＋Tyr203His＋Pro204Gln＋Tyr208Leu＋Gly213ProAsn198Gln＋Thr202Asn＋Tyr203Ala＋Pro204Glu＋Tyr208Gly＋Ala209Gln＋Thr214GlnAla194Asn＋Pro195Gln＋Pro204Gly＋Ser206Glu＋Thr207Ser＋Tyr208Ala＋Asn212SerAla194Pro＋Pro195Gln＋Val197Ser＋Asn198Ser＋Gln200Glu＋Thr202Ser＋Gly205SerPro195Ser＋Gly196Ser＋Val199Gly＋Thr202Gly＋Pro204Glu＋Leu211Ala＋Thr214AsnAla194Thr＋Gly196Pro＋Asn198Ser＋Pro204Asn＋Tyr208Gly＋Ser210Asp＋Thr214AsnAla194Pro＋Val199Asn＋Thr202Gln＋Pro204Gln＋Gly205Asp＋Thr207Ser＋Ala209ProAla194Pro＋Pro195Gly＋Val197Cys＋Val199Gln＋Tyr203Cys＋Tyr208His＋Gly213GluVal197Asp＋Val199Ser＋Gly205Pro＋Thr207Gly＋Leu211Thr＋Asn212Gln＋Gly213SerAla194Pro＋Pro195Ser＋Gln200Asn＋Thr202Pro＋Tyr208His＋Gly213Asp＋Thr214GlyAla194Thr＋Val197Asn＋Val199Pro＋Gln200Asp＋Thr202Ser＋Thr207Asn＋Tyr208LeuPro195Ser＋Gly196Gln＋Val197Ala＋Thr202Gln＋Tyr203Gly＋Tyr208Cys＋Gly213AsnAla194Gln＋Val199Gln＋Thr202Gly＋Pro204Asn＋Gly205Glu＋Ala209His＋Leu211HisPro195Asn＋Val199Thr＋Tyr203Gly＋Gly205Gln＋Ser206Asp＋Asn212Ser＋Gly213ProAla194Gln＋Pro195Ser＋Gly196Gln＋Thr202Gln＋Ala209His＋Asn212Ser＋Thr214GluGly196Pro＋Val197Cys＋Tyr203Pro＋Gly205Ser＋Ser206Asp＋Leu211Ile＋Thr214GlyGly196Asn＋Val197Gly＋Val199His＋Thr202Ser＋Ala209His＋Asn212Glu＋Thr214GlnGly196Gln＋Gln200Ser＋Thr202Gln＋Gly205Gln＋Ala209Ser＋Gly213Asp＋Thr214GluVal199Gly＋Gln200Asn＋Thr202Gln＋Gly205Ser＋Ala209Glu＋Ser210Asp＋Leu211AsnVal199Pro＋Gln200Asn＋Tyr203Val＋Ala209Asp＋Ser210Asp＋Leu211Ser＋Gly213ProAla194Gly＋Tyr203Val＋Gly205Asn＋Thr207Gly＋Ala209Glu＋Ser210Asp＋Leu211ProPro195Gln＋Thr202Gly＋Tyr203Val＋Ala209Glu＋Ser210Glu＋Gly213Ser＋Thr214GlnPro195Asn＋Val197Ser＋Tyr208Thr＋Ala209Glu＋Ser210Asp＋Asn212Ser＋Thr214GlnAla194Asn＋Gly196Ser＋Val199Gly＋Thr202Pro＋Tyr203Leu＋Asn212Glu＋Gly213AspVal197Cys＋Asn198Ser＋Gln200Ser＋Thr202Asn＋Thr207Gln＋Asn212Asp＋Gly213AspGly196Pro＋Thr202Asn＋Pro204Asp＋Gly205Glu＋Ala209His＋Leu211His＋Gly213SerPro195Gly＋Gly196Gln＋Tyr203Thr＋Thr207Asn＋Ser210Asp＋Leu21lAsp＋Asn212SerAla194Gly＋Gly196Pro＋Thr202Gln＋Tyr203Val＋Gly205Ser＋Ser210Asp＋Leu211AspAla194His＋Val199Cys＋Tyr203Ala＋Tyr208Gly＋Ser210Asp＋Leu211Asp＋Gly213ProAla194His＋Gly196Ser＋Val197Cys＋Val199Ser＋Gly205Glu＋Ser206Glu＋Tyr208SerAla194His＋Gly196Ser＋Val197Thr＋Val199Cys＋Gly205Asp＋Ser206Glu＋Thr214GlnThr202Asn＋Tyr203Thr＋Gly205Glu＋Ser206Asp＋Ala209Gln＋Leu211Thr＋Thr214GlyAla194Pro＋Val197Met＋Asn198Ser＋Val199Asn＋Thr202Ser＋Ser206Asp＋Thr207GluPro195Asn＋Val199Met＋Thr202Gly＋Ser206Glu＋Thr207Asp＋Tyr208Thr＋Asn212SerPro195Gly＋Gly196Asn＋Val199Gln＋Tyr203His＋Ser206Glu＋Thr207Asp＋Asn212GlnPro195Asn＋Val199Ala＋Gly205Pro＋Ser206Glu＋Thr207Asp＋Tyr208Gly＋Asn212SerPro195Asn＋Gly196Ser＋Val197Ala＋Pro204Ser＋Ser206Glu＋Thr207Asp＋Asn212GlnPro195Asn＋Val197Thr＋Ser206Asp＋Thr207Asp＋Leu211Thr＋Gly213Ser＋Thr214SerAla194Thr＋Val197Cys＋Asn198Ser＋Thr207Asp＋Tyr208Asp＋Asn212Ser＋Gly213AsnAla194Thr＋Gln200Asp＋Thr202Gln＋Ala209His＋Ser210Asp＋Leu211Gly＋Thr214GlyPro195Gln＋Gln200Asp＋Tyr203Ala＋Gly205Asn＋Tyr208Ser＋Ser210Asp＋Gly213AsnAla194Thr＋Val199Pro＋Gln200Glu＋Thr207Gly＋Ser210Glu＋Leu211Ser＋Gly213AsnVal197Ser＋Gln200Glu＋Tyr203Leu＋Pro204Asn＋Ala209Gly＋Ser210Asp＋Leu211AsnGly196Gln＋Asn198Gln＋Val199Pro＋Gln200Asp＋Thr202Gly＋Pro204Gly＋Ser210AspAsn198Gln＋Val199Cys＋Gln200Glu＋Gly205Gln＋Ser210Asp＋Gly213Ser＋Thr214GlyAsn198Gln＋Gln200Glu＋Gly205Pro＋Thr207Ser＋Ala209Asn＋Ser210Glu＋Thr214AsnGln200Glu＋Thr202Asn＋Thr207Ser＋Tyr208Pro＋Ser210Glu＋Gly213Gln＋Thr214GlyAla194Pro＋Pro195Ser＋Gly196Asn＋Val197Asp＋Val199Thr＋Pro204Gly＋Asn212GluGly196Asn＋Asn198Glu＋Tyr203Gly＋Pro204Gln＋Ser210Asp＋Leu211Val＋Asn212SerGly196Pro＋Val197Gly＋Asn198Asp＋Thr202Pro＋Tyr203Val＋Thr207Asn＋Ser210GluPro195Asn＋Gly196Pro＋Asn198Asp＋Thr202Ser＋Pro204Gln＋Ser210Asp＋Gly213GlnPro195Ser＋Val197Pro＋Asn198Glu＋Val199Met＋Thr207Ser＋Ser210Asp＋Thr214GlnAla194Ser＋Gly196Asn＋Val197Met＋Asn198Asp＋Val199Thr＋Tyr208Met＋Leu211GluAsn198Asp＋Val199Gln＋Pro204Ser＋Leu211Asp＋Asn212Ser＋Gly213Pro＋Thr214AsnPro195Gln＋Asn198Asp＋Thr202Pro＋Gly205Gln＋Tyr208Met＋Leu211Glu＋Gly213AsnGly196Pro＋Gln200Asp＋Thr202Ser＋Tyr203Thr＋Tyr208His＋Ala209Glu＋Leu211IleAla194Thr＋Gln200Asn＋Thr202Pro＋Pro204Asp＋Ser206Glu＋Thr207Asn＋Ala209SerGly196Pro＋Pro204Asp＋Gly205Ser＋Ser206Asp＋Tyr208Gly＋Gly213Gln＋Thr214GlyPro195Gln＋Val197Gln＋Gln200Ser＋Pro204Glu＋Ser206Asp＋Ala209Gly＋Thr214GlnAla194Thr＋Pro195Gly＋Val197Pro＋Pro204Asp＋Ser206Glu＋Leu211Val＋Gly213SerPro195Asn＋Gly196Ser＋Pro204Glu＋Ser206Asp＋Thr207Gly＋Tyr208Val＋Ala209ThrPro195Asn＋Gly196Pro＋Val197Gln＋Asn198Asp＋Tyr203Gln＋Tyr208Asn＋Asn212AspGly196Gln＋Val197Thr＋Asn198Asp＋Val199Gly＋Thr202Pro＋Tyr208Asn＋Asn212GluPro195Asn＋Asn198Asp＋Val199Asn＋Thr202Gln＋Ala209Pro＋Asn212Asp＋Gly213GlnAla194Gly＋Val197Met＋Asn198Asp＋Gln200Glu＋Thr202Asn＋Pro204Asn＋Tyr208AsnPro195Ser＋Val197Gln＋Asn198Glu＋Val199Cys＋Gln200Asp＋Gly205Ser＋Ala209ProPro195Gln＋Gly196Pro＋Asn198Glu＋Val199Ser＋Gln200Asp＋Gly205Ser＋Thr214AsnAla194Gln＋Pro195Ser＋Asn198Glu＋Gln200Asp＋Thr207Gln＋Tyr208Leu＋Asn212GlnAla194Pro＋Pro195Asn＋Gln200Glu＋Pro204Gln＋Thr207Gln＋Tyr208Asp＋Gly213GlnVal199Thr＋Gln200Asp＋Tyr208Glu＋Ala209Thr＋Leu211Ser＋Asn212Ser＋Gly213AsnGly196Pro＋Val197Glu＋Asn198Gln＋Val199Met＋Tyr208Met＋Gly213Asp＋Thr214AsnPro195Gly＋Val197Glu＋Val199Thr＋Gln200Ser＋Gly205Asn＋Ala209Gly＋Leu211GluVal197Met＋Val199Cys＋Gly205Pro＋Tyr208Ile＋Leu211Asp＋Asn212Ser＋Gly213AspPro195Gln＋Asn198Ser＋Val199Gln＋Pro204Ser＋Ser210Asp＋Leu211Cys＋Asn212GluPro195Ser＋Gly196Pro＋Asn198Gln＋Pro204Gln＋Ala209Thr＋Ser210Glu＋Asn212GluAla194Gly＋Asn198Gln＋Val199Gln＋Thr207Ser＋Ser210Asp＋Leu211Gly＋Asn212AspAla194Gln＋Val197Gly＋Thr202Ser＋Pro204Ser＋Ser210Glu＋Asn212Glu＋Gly213SerAla194Asn＋Asn198Ser＋Thr202Pro＋Ser210Glu＋Leu211Cys＋Asn212Asp＋Gly213SerPro195Gly＋Val197Thr＋Thr202Asn＋Gly205Ser＋Ser210Asp＋Asn212Glu＋Gly213SerPro195Gln＋Gly205Gln＋Ala209Gln＋Ser210Asp＋Leu21lAsn＋Asn212Glu＋Thr214GlnVal199His＋Pro204Asn＋Tyr208Ile＋Ala209Thr＋Ser210Glu＋Leu211Gln＋Asn212AspVal197Asn＋Asn198Gln＋Tyr203Pro＋Pro204Asn＋Ser210Glu＋Asn212Glu＋Thr214GlnAla194Pro＋Gly196Ser＋Gln200Ser＋Gly205Gln＋Tyr208Gly＋Ser210Glu＋Asn212GluPro195Ser＋Val197Gly＋Asn198Gln＋Val199Gln＋Gln200Ser＋Ser210Glu＋Asn212AspAsn198Gln＋Gln200Asn＋Thr207Gly＋Tyr208Glu＋Ser210Glu＋Asn212Ser＋Thr214AsnPro195Gly＋Thr202Ser＋Tyr208Glu＋Ala209Asn＋Ser210Glu＋ Asn212Gln＋Thr214AsnVal199Thr＋Thr202Ser＋Gly205Pro＋Ser206Glu＋Tyr208Glu＋Leu211His＋Asn212SerAla194Gln＋Gly196Pro＋Asn198Gln＋Tyr203Ile＋Ser206Asp＋Tyr208Asp＋Gly213GlnAla194Ser＋Pro195Ser＋Thr202Ser＋Gly205Gln＋Ser206Asp＋Tyr208Asp＋Asn212GlnPro195Gly＋Val197His＋Tyr203Gln＋Thr207Pro＋Asn212Asp＋Gly213Ser＋Thr214AspPro195Gly＋Val197Met＋Thr202Asn＋Tyr208Met＋Leu211Thr＋Asn212Glu＋Thr214AspVal197Ser＋Asn198Gln＋Thr202Ser＋Pro204Glu＋Thr207Ser＋Tyr208Asp＋Ala209SerAla194Pro＋Val199Asn＋Pro204Asp＋Tyr208Glu＋Leu211Gly＋Asn212Gln＋Gly213SerPro195Gln＋Val197Glu＋Val199Gly＋Gln200Ser＋Thr207Ser＋Ser210Glu＋Leu211AsnPro195Ser＋Val197Asp＋Val199Cys＋Pro204Gly＋Tyr208Ser＋Ser210Glu＋Asn212SerAla194Gln＋Val197Asp＋Asn198Ser＋Thr202Asn＋Thr207Asn＋Ser210Glu＋Leu211HisVal197Asp＋Gln200Asn＋Tyr203Thr＋Tyr208Val＋Ser210Asp＋Asn212Ser＋Thr214SerAla194Pro＋Val197Asp＋Gln200Ser＋Thr202Gly＋Leu211Cys＋Gly213Asn＋Thr214GluPro195Ser＋Val197Glu＋Asn198Gln＋Tyr208Ile＋Ala209His＋Leu211Val＋Thr214GluPro195Gly＋Val197Asp＋Gln200Ser＋Thr202Pro＋Ala209Thr＋Gly213Gln＋Thr214AspPro195Asn＋Gly196Pro＋Asn198Asp＋Pro204Asn＋Gly205Asn＋Ala209Asn＋Gly213AspAla194Pro＋Asn198Asp＋Val199Gln＋Gln200Ser＋Thr207Pro＋Ala209Asp＋Gly213AsnVal197Asp＋Val199His＋Gln200Glu＋Thr202Pro＋Tyr203Ile＋Thr207Gly＋Thr214GlyVal197Glu＋Gln200Asp＋Tyr203Ile＋Pro204Gln＋Ala209Gly＋Leu211Ile＋Thr214ProPro195Asn＋Gly1965er＋Val199Met＋Gln200Asp＋Tyr203Ala＋Asn212Asp＋Gly213AsnAla194Thr＋Gly196Gln＋Gln200Glu＋Tyr203Val＋Tyr208Ala＋Leu211Ala＋Asn212GluVal197Asn＋Asn198Gln＋Pro204Asp＋Tyr208Met＋Ala209Glu＋Asn212Ser＋Gly213AsnPro195Asn＋Thr202Gly＋Tyr203Asn＋Pro204Glu＋Gly205Asn＋Thr207Ser＋Ala209GluAla194Pro＋Gly196Pro＋Tyr203Ala＋Thr207Gln＋Leu211Glu＋Gly213Pro＋Thr214GluAsn198Gln＋Val199Thr＋Thr202Pro＋Pro204Gln＋Ala209Thr＋Leu211Glu＋Thr214AspAla194His＋Val199Cys＋Gln200Glu＋Tyr203Ile＋Gly205Asn＋Thr207Glu＋Tyr208MetVal199Pro＋Gln200Asp＋Pro204Gln＋Gly205Asn＋Thr207Asp＋Tyr208Cys＋Thr214ProAla194His＋Pro195Gln＋Gly196Gln＋Val199His＋Gln200Glu＋Thr207Glu＋Asn212SerAla194Gly＋Asn198Gln＋Gln200Asn＋Tyr203Leu＋Ser210Asp＋Leu211Gly＋Gly213GluAsn198Gln＋Gln200Asn＋Thr202Ser＋Ser210Asp＋Leu211Ser＋Gly213Glu＋Thr214AsnPro195Asn＋Val197Met＋Tyr203Gly＋Gly205Pro＋Tyr208His＋Ser210Asp＋Gly213AspPro195Ser＋Gln200Asn＋Thr207Pro＋Ser210Glu＋Leu211Met＋Gly213Glu＋Thr214GlnVal197Gln＋Val199Ser＋Thr202Asn＋Tyr208Gly＋Ala209Gly＋Ser210Glu＋Gly213AspGln200Asn＋Pro204Gln＋Thr207Glu＋Ala209Ser＋Ser210Asp＋Leu211Thr＋Gly213AsnAla194Gln＋Thr202Gly＋Tyr203Asn＋Thr207Asp＋Ser210Asp＋Leu211Cys＋Gly213ProPro195Gln＋Val199Gln＋Gly205Gln＋Thr207Glu＋Tyr208Ile＋Ser210Asp＋Thr214GlyPro195Gly＋Val197His＋Val199Met＋Tyr203Thr＋Thr207Glu＋Ser210Asp＋Asn212GlnAla194Gln＋Pro195Gln＋Asn198Ser＋Val199Gln＋Pro204Gln＋Thr207Glu＋Ser210GluVal197Pro＋Asn198Glu＋Val199Thr＋Pro204Asn＋Leu211Cys＋Asn212Ser＋Thr214AspAla194Ser＋Val199Thr＋Gln200Glu＋Tyr203Cys＋Tyr208Gly＋Leu211Gly＋Gly213GluVal199Pro＋Gln200Asp＋Tyr203Asn＋Pro204Gly＋Leu211Pro＋Gly213Glu＋Thr214AsnAla194Gln＋Asn198Glu＋Thr207Ser＋Tyr208Glu＋Asn212Gln＋Gly213Gln＋Thr214GlnPro195Gln＋Asn198Asp＋Gln200Ser＋Gly205Ser＋Tyr208Asp＋Leu211Ala＋Asn212SerGly196Pro＋Val197Asp＋Asn198Gln＋Val199Ala＋Pro204Gln＋Ala209Asp＋Gly213AsnPro195Ser＋Asn198Gln＋Val199Gly＋pro204Glu＋Ala209Gly＋Ser210Asp＋Asn212GlnPro195Ser＋Gln200Ser＋Pro204Glu＋Thr207Pro＋Ala209Gln＋Ser210Asp＋Asn212GlnAla194His＋Gly196Ser＋Val197Gly＋Val199Asn＋Gln200Glu＋Pro204Glu＋Ala209ThrGly196Pro＋Asn198Ser＋Pro204Asn＋Tyr208Gly＋Ala209Asp＋Leu211Thr＋Gly213AspGly196Pro＋Val199Thr＋Gln200Asp＋Thr202Asn＋Tyr203Asn＋Ser206Glu＋Ala209SerPro195Ser＋Gly196Pro＋Val199Pro＋Gln200Glu＋Pro204Gln＋Ser206Asp＋Leu211AlaPro195Ser＋Gln200Asp＋Thr202Gln＋Pro204Gln＋Ser206Glu＋Tyr208Gly＋Ala209GlyGly196Asn＋Val199Ser＋Gln200Glu＋Thr202Pro＋Ala209Thr＋Leu211Cys＋Thr214AspVal197Asn＋Gln200Glu＋Tyr203Gln＋Gly205Pro＋Ala209Thr＋Leu211Val＋Thr214GluPro195Gly＋Tyr203Gly＋Ser206Glu＋Tyr208Gly＋Ala209Gln＋Ser210Glu＋Leu211IleGly196Asn＋Val197His＋Val199Ala＋Pro204Asn＋Ser206Asp＋Ser210Asp＋Leu211ThrAla194Gln＋Val197Ser＋Val199Pro＋Ser206Asp＋Ser210Glu＋Leu211Pro＋Thr214GlyAla194His＋Gln200Ser＋Thr202Asn＋Ser206Glu＋Tyr208Cys＋Ser210Asp＋Asn212SerVal197Thr＋Thr202Ser＋Pro204Gly＋Gly205Asn＋Ser206Asp＋Ser210Glu＋Leu211Pro表30第6环状区-八突变的突变体Pro195Ser＋Val197Ser＋Asn198Ser＋Val199Gly＋Tyr203Thr＋Thr207Asn＋Tyr208Met＋Asn212GluPro195Gln＋Gly196Gln＋Val199Gln＋Tyr203Gln＋Tyr208Met＋Ala209Gly＋Leu211Val＋Gly213AspPro195Gln＋Asn198Gln＋Val199His＋Pro204Gly＋Ser206Glu＋Thr207Gln＋Tyr208Ser＋Leu211GlnAla194Thr＋Val197Met＋Val199His＋Thr202Ser＋Tyr203Ile＋Thr207Gln＋Tyr208Pro＋Leu211GlyPro195Gln＋Val197Asn＋Val199Cys＋Tyr203Met＋Pro204Asn＋Gly205Glu＋Thr207Pro＋Tyr208SerGly196Ser＋Gln200Asn＋Pro204Asp＋Gly205Ser＋Thr207Pro＋Tyr208Met＋Gly213Gln＋Thr214SerVal197Gln＋Gln200Asn＋Tyr203His＋Pro204Asp＋Gly205Ser＋Tyr208Thr＋Ala209Gln＋Leu211ProPro195Gly＋Gly196Ser＋Val197Pro＋Val199Ser＋Thr202Asn＋Ala209Glu＋Leu211Ser＋Gly213ProAla194His＋Gly196Ser＋Val197Asn＋Asn198Ser＋Tyr203Leu＋Pro204Asn＋Thr207Ser＋Ala209ThrAla194Pro＋Gly196Pro＋Val199Met＋Gln200Asp＋Thr202Asn＋Thr207Asn＋Ala209Pro＋Gly213ProPro195Asn＋Gly196Pro＋Val199Thr＋Thr202Gln＋Thr207Gly＋Tyr208Ser＋Ser210Asp＋Asn212SerAla194Thr＋Pro195Asn＋Gly196Gln＋Thr202Pro＋Pro204Ser＋Gly205Pro＋Tyr208Gln＋Leu211AspAla194Gly＋Pro195Ser＋Val197Ala＋Thr202Gly＋Tyr203Gly＋Pro204Gln＋Gly205Ser＋Leu211GluPro195Asn＋Val197Glu＋Val199Thr＋Gln200Asn＋Thr202Pro＋Tyr203Ile＋Pro204Ser＋Leu211CysAla194His＋Pro195Gln＋Val199Cys＋Gln200Asp＋Tyr203Ala＋Thr207Ser＋Gly213Gln＋Thr214ProAla194Gln＋Gly196Gln＋Val199His＋Thr202Pro＋Tyr203Thr＋Pro204Asp＋Gly205Ser＋Ala209AsnPro195Ser＋Gly196Ser＋Gln200Asn＋Tyr203His＋Thr207Pro＋Leu211Met＋Gly213Glu＋Thr214GlnVal197Cys＋Asn198Ser＋Val199Ala＋Gln200Asp＋Tyr203His＋Leu211Asn＋Asn212Gln＋Gly213AsnAla194Gln＋Gly196Pro＋Asn198Gln＋Val199His＋Tyr203Val＋Thr207Ser＋Ala209Ser＋Leu211AsnPro195Gly＋Gly196Ser＋Val197Pro＋Thr207Gly＋Leu211Thr＋Asn212Ser＋Gly213Asn＋Thr214GluVal197Asn＋Thr202Gly＋Tyr203Ala＋Gly205Gln＋Tyr208Ile＋Ala209Thr＋Ser210Asp＋Leu211GlyAla194Asn＋Asn198Gln＋Val199Ser＋Thr202Gly＋Tyr203Ile＋Ala209Thr＋Ser210Asp＋Thr214SerPro195Ser＋Gly196Asn＋Val197Asn＋Asn198Ser＋Thr202Asn＋Tyr203Cys＋Tyr208Asn＋Leu211ThrAla194Gly＋Pro195Ser＋Gly196Gln＋Val199Ala＋Thr202Pro＋Thr207Ser＋Asn212Ser＋Thr214AspVal197Cys＋Val199Pro＋Thr202Pro＋Tyr203Ser＋Ser206Glu＋Thr207Gln＋Ala209Gly＋Thr214GlyAla194Gly＋Pro195Gln＋Gly196Gln＋Asn198Ser＋Val199Met＋Thr202Pro＋Thr207Glu＋Thr214GlyAla194His＋Pro195Gln＋Val197Gln＋Asn198Gln＋Tyr203Ser＋Ser206Glu＋Tyr208Ala＋Leu211AlaAla194Gly＋Gly196Ser＋Tyr203His＋Pro204Asn＋Ser206Asp＋Tyr208Ile＋Leu211Gln＋Asn212SerGly196Pro＋Gln200Ser＋Tyr203Gln＋Pro204Ser＋Gly205Ser＋Tyr208Cys＋Leu211Met＋Gly213GlnAla194Asn＋Tyr203Asn＋Gly205Glu＋Thr207Gln＋Ala209Thr＋Leu211His＋Gly213Pro＋Thr214GlnAla194Ser＋Pro195Asn＋Gly196Asn＋Val199Ser＋Pro204Ser＋Gly205Asn＋Ala209Asp＋Thr214GlnPro195Asn＋Gly196Gln＋Thr202Pro＋Gly205Ser＋Ser206Glu＋Tyr208Ser＋Ala209Pro＋Gly213GlnAla194Thr＋Pro195Gly＋Val197Asp＋Asn198Gln＋Gly205Ser＋Tyr208Val＋Asn212Gln＋Gly213SerPro195Gly＋Asn198Ser＋Tyr203Thr＋Pro204Asn＋Tyr208Pro＋Ala209Thr＋Asn212Ser＋Gly213SerAla194Thr＋Pro195Ser＋Gly196Asn＋Val197Met＋Gln200Ser＋Thr202Asn＋Tyr203His＋Tyr208GlnAla194His＋Asn198Ser＋Tyr203Gln＋Pro204Gln＋Ala209Pro＋Leu211Asp＋Asn212Glu＋Gly213AsnAla194Asn＋Pro195Ser＋Val197Gly＋Pro204Gly＋Gly205Asn＋Ala209Asp＋Ser210Asp＋Thr214GlnGly196Gln＋Val197Pro＋Gln200Asn＋Thr202Asn＋Gly205Gln＋Ala209Glu＋Ser210Asp＋Thr214ProVal197Ser＋Val199Ala＋Thr202Ser＋Tyr203Pro＋Gly205Asn＋Tyr208Ile＋Ala209Asp＋Ser210GluAsn198Ser＋Val199Met＋Gln200Ser＋Pro204Ser＋Tyr208Ala＋Ala209Glu＋Ser210Asp＋Leu211GlyAla194Gly＋Val197Asn＋Asn198Ser＋Gln200Ser＋Tyr203Thr＋Ala209Asp＋Ser210Asp＋Thr214ProAla194His＋Pro195Ser＋Asn198Ser＋Tyr203Cys＋Thr207Ser＋Ala209Asp＋Ser210Glu＋Gly213SerPro195Ser＋Val199Asn＋Gln200Ser＋Pro204Ser＋Thr207Gly＋Ala209Asp＋Ser210Asp＋Asn212SerAla194Gln＋Gly196Pro＋Val197Asp＋Asn198Glu＋Val199Ser＋Gly205Gln＋Ala209Gly＋Asn212SerPro195Gln＋Val197Asp＋Asn198Glu＋Val199Ala＋Gln200Ser＋Thr202Ser＋Thr207Gly＋Tyr208AlaPro195Ser＋Gly196Pro＋Gln200Asn＋Thr202Asn＋Tyr203Met＋Thr207Gly＋Asn212Asp＋Gly213AspGly196Asn＋Thr202Pro＋Pro204Gly＋Thr207Pro＋Leu211Cys＋Asn212Glu＋Gly213Glu＋Thr214ProGly196Asn＋Asn198Gln＋Gln200Ser＋Thr202Pro＋Tyr203Cys＋Pro204Asn＋Asn212Asp＋Gly213GluAsn198Ser＋Val199Gly＋Tyr203Ala＋Pro204Asp＋Gly205Glu＋Tyr208Met＋Ala209Ser＋Leu211ProVal197Cys＋Val199Asn＋Thr202Gln＋Pro204Asp＋Gly205Asp＋Ala209Gln＋Leu211Gln＋Asn212SerPro195Asn＋Val199Thr＋Thr202Gly＋Ala209His＋Ser210Glu＋Leu211Asp＋Asn212Ser＋Gly213AsnGly196Asn＋Val199His＋Tyr203Ala＋Gly205Asp＋Ser206Asp＋Thr207Gln＋Tyr208Gln＋Leu211AsnAla194Gly＋Gly196Gln＋Val199Gln＋Gln200Asn＋Pro204Gly＋Gly205Asp＋Ser206Asp＋Thr214SerAla194Gln＋Pro195Asn＋Val197Ala＋Gln200Asn＋Tyr203Ser＋Gly205Asp＋Ser206Asp＋Tyr208AlaAla194Gln＋Val197Gln＋Pro204Asn＋Gly205Asp＋Ser206Glu＋Tyr208Ala＋Ala209His＋Gly213AsnPro195Ser＋Gly196Asn＋Val197Thr＋Gln200Asn＋Pro204Ser＋Gly205Glu＋Ser206Glu＋Tyr208GlyVal197Gly＋Asn198Gln＋Val199Met＋Gln200Asn＋Tyr203His＋Ser206Asp＋Thr207Glu＋Thr214ProAla194Gln＋Pro195Gly＋Gly196Pro＋Thr202Pro＋Ser206Asp＋Thr207Asp＋Tyr208Val＋Ala209HisAla194Thr＋Pro195Ser＋Val197Gln＋Asn198Gln＋Thr202Ser＋Pro204Asn＋Ser206Glu＋Thr207AspAla194Gln＋Val199Met＋Thr202Ser＋Tyr203Gly＋Gly205Gln＋Ser206Asp＋Thr207Glu＋Gly213SerAla194Gly＋Pro195Asn＋Gly196Gln＋Tyr203Ala＋Ser206Asp＋Thr207Glu＋Ala209Thr＋Asn212GlnGly196Ser＋Gln200Ser＋Tyr203Leu＋Pro204Ser＋Ser206Glu＋Thr207Glu＋Ala209Ser＋Gly213SerAla194Ser＋Pro195Asn＋Thr202Gln＋Ser206Glu＋Thr207Glu＋Tyr208Gly＋Asn212Ser＋Thr214GlyAla194His＋Pro195Gln＋Val197Ala＋Thr202Asn＋Tyr203Pro＋Gly205Pro＋Tyr208Asp＋Ala209AspAla194Gln＋Pro195Gln＋Val197Asn＋Gln200Asp＋Ala209Gly＋Ser210Glu＋Leu211Ala＋Thr214AsnAla194Pro＋Val199Met＋Gln200Glu＋Thr202Gly＋Pro204Gln＋Gly205Asn＋Tyr208Gln＋Ser210AspGly196Pro＋Val197Cys＋Val199Asn＋Gln200Glu＋Gly205Pro＋Tyr208Val＋Ser210Glu＋Thr214ProAla194Thr＋Pro195Gly＋Gly196Gln＋Val197Asp＋Val199His＋Gln200Ser＋Thr202Ser＋Asn212AspAsn198Asp＋Val199Pro＋Gly205Asn＋Thr207Gly＋Tyr208Pro＋Ser210Asp＋Leu211Gly＋Gly213GlnAla194Gln＋Gly196Pro＋Asn198Glu＋Tyr203Gln＋Thr207Gln＋Tyr208Pro＋Ala209Pro＋Ser210GluAla194Pro＋Asn198Asp＋Val199His＋Gly205Asn＋Ser210Glu＋Leu211Asn＋Asn212Ser＋Thr214ProAsn198Glu＋Val199Pro＋Tyr203Asn＋Thr207Pro＋Ala209Asn＋Ser210Glu＋Asn212Ser＋Gly213GlnAla194Gly＋Pro195Asn＋Asn198Asp＋Tyr203Ala＋Gly205Gln＋Thr207Ser＋Ser210Glu＋Leu211ValAla194Thr＋Asn198Asp＋Thr202Gly＋Pro204Ser＋Gly205Ser＋Thr207Asn＋Ala209Pro＋Ser210GluAla194Gln＋Asn198Asp＋Tyr203Ala＋Thr207Gln＋Tyr208Pro＋Ser210Glu＋Leu211Pro＋Asn212SerAla194Thr＋Pro195Ser＋Asn198Asp＋Tyr203Cys＋Pro204Asn＋Thr207Gln＋Tyr208Gln＋Ser210GluPro195Ser＋Asn198Asp＋Val199Ser＋Gln200Ser＋Pro204Ser＋Ser210Glu＋Leu211Gly＋Asn212SerAla194Gln＋Pro195Ser＋Gly196Pro＋Gln200Glu＋Pro204Asn＋Gly205Pro＋Ala209Glu＋Leu211CysVal199His＋Gln200Ser＋Pro204Asp＋Gly205Ser＋Ser206Asp＋Tyr208Gly＋Gly213Gln＋Thr214GlyGly196Asn＋Asn198Ser＋Thr202Ser＋Pro204Asp＋Gly205Ser＋Ser206Asp＋Thr207Ser＋Tyr208ValVal197Ala＋Gln200Asn＋Pro204Glu＋Gly205Ser＋Ser206Glu＋Tyr208Val＋Leu211Ser＋Asn212SerPro195As＋Val197Thr＋Thr202Ser＋Tyr203Ile＋Pro204Asp＋Ser206Asp＋Thr207Gln＋Ala209GlnPro195Gly＋Val197Asn＋Val199Ser＋Gln200Asn＋Tyr203Asn＋Pro204Glu＋Ser206Glu＋Leu211MetGly196Ser＋Asn198Ser＋Val199Met＋Gln200Asn＋Pro204Glu＋Ser206Asp＋Leu211Val＋Thr214GlnAla194Gln＋Gly196Asn＋Asn198Asp＋Gln200Ser＋Thr202Asn＋Pro204Asn＋Asn212Glu＋Thr214AsnPro195Gly＋Asn198Glu＋Val199Asn＋Gln200Ser＋Thr202Gly＋Thr207Asn＋Ala209Thr＋Asn212AspPro195Asn＋Gly196Ser＋Asn198Glu＋Thr207Ser＋Tyr208Ser＋Ala209Pro＋Asn212Asp＋Thr214AsnAla194Pro＋Val197Ser＋Asn198Glu＋Gln200Asn＋Tyr203Cys＋Thr207Pro＋Tyr208Thr＋Asn212AspPro195Gly＋Gly196Ser＋Thr202Asn＋Pro204Asn＋Gly205Glu＋Thr207Asp＋Tyr208Thr＋Asn212SerVal197Gln＋Val199Asn＋Thr202Pro＋Gly205Glu＋Thr207Glu＋Tyr208Ala＋Leu211Ser＋Gly213AsnAla194Ser＋Pro195Ser＋Val197Gly＋Pro204Asp＋Gly205Ser＋Thr207Glu＋Tyr208Val＋Leu211GlyAla194Gln＋Pro195Gly＋Gly196Asn＋Val197Cys＋Asn198Gln＋Pro204Glu＋Thr207Asp＋Gly213GlnPro195Gly＋Asn198Asp＋Gln200Glu＋Tyr203Thr＋Gly205Asn＋Leu211Cys＋Asn212Ser＋Gly213GlnAla194Gly＋Pro195Asn＋Asn198Glu＋Gln200Glu＋Thr202Gln＋Gly205Ser＋Thr207Pro＋Gly213GlnPro195Gly＋Asn198Glu＋Val199Gln＋Gln200Asp＋Thr202Gly＋Pro204Gln＋Thr207Pro＋Leu211CysPro195Asn＋Gly196Gln＋Asn198Glu＋Gln200Asp＋Thr202Gly＋Tyr208Gln＋Leu21lSer＋Thr214SerVal197Asn＋Asn198Ser＋Gln200Glu＋Thr202Asn＋Tyr203Cys＋Pro204Ser＋Gly205Pro＋Tyr208GluAla194His＋Val199His＋Tyr203Asn＋Tyr208His＋Ala209Glu＋Leu211Asp＋Asn212Gln＋Gly213GlnGly196Gln＋Val197Ser＋Gln200Glu＋Tyr203Val＋Thr207Pro＋Ala209Thr＋Leu211Glu＋Asn212SerPro195Gln＋Val199Gly＋Gln200Asp＋Pro204Asn＋Gly205Asn＋Tyr208Ile＋Ala209Pro＋Leu211GluPro195Gln＋Asn198Ser＋Val199Asn＋Gln200Asp＋Gly205Pro＋Tyr208Ser＋Leu211Glu＋Thr214AsnAla194His＋Pro195Ser＋Val197Glu＋Val199Cys＋Thr202Pro＋Leu211Glu＋Asn212Gln＋Thr214AsnAla194Ser＋Pro195Gln＋Asn198Ser＋Gln200Asn＋Gly205Gln＋Leu211Asp＋Asn212Gln＋Gly213GluGly196Gln＋Val199Gln＋Tyr203Gly＋Gly205Asn＋Ala209His＋Leu211Asp＋Asn212Gln＋Gly213AspAsn198Gln＋Gln200Ser＋Tyr203Leu＋Gly205Asn＋Thr207Glu＋Ala209Asp＋Leu211Ser＋Gly213AsnAla194Ser＋Pro195Ser＋Val199Gln＋Gln200Ser＋Gly205Gln＋Ser210Asp＋Asn212Glu＋Thr214GlnPro195Asn＋Tyr203Cys＋Pro204Gln＋Tyr208Asn＋Ala209His＋Ser210Asp＋Asn212Asp＋Thr214GlyGly196Asn＋Val199Ser＋Thr202Gln＋Pro204Ser＋Thr207Gly＋Ser210Glu＋Leu211Thr＋Asn212AspGly196Gln＋Val197Cys＋Asn198Ser＋Pro204Gln＋Thr207Asn＋Ala209Gln＋Ser210Asp＋Asn212AspPro195Gly＋Gly196Pro＋Val197Cys＋Asn198Ser＋Gln200Asn＋Tyr208His＋Ser210Asp＋Asn212GluPro195Gln＋Asn198Ser＋Val199Gly＋Thr202Pro＋Thr207ser＋Tyr208Ile＋Ser210Asp＋Asn212AspAla194Asn＋Val197Met＋Val199Thr＋Ala209Asn＋Ser210Asp＋Leu211Val＋Asn212Glu＋Gly213GlnAla194Gln＋Pro195Asn＋Val199Asn＋Tyr203Ser＋Thr207Asn＋Ser210Asp＋Leu211Gly＋Asn212AspVal199Cys＋Gln200Ser＋Thr202Asn＋Gly205Asn＋Thr207Gly＋Tyr208Asn＋Ser210Glu＋Asn212AspAla194Thr＋Val199Cys＋Gln200Ser＋Gly205Ser＋Ser206Glu＋Tyr208Asp＋Leu211Ala＋Gly213AsnAla194Pro＋Val197Pro＋Gln200Ser＋Ser206Asp＋Thr207Pro＋Tyr208Asp＋Ala209Pro＋Thr214GlyGly196Gln＋Asn198Ser＋Val199Met＋Thr202Ser＋Pro204Asn＋Ser206Glu＋Tyr208Glu＋Asn212GlnPro195Asn＋Gly196Asn＋Val199Ala＋Gln200 ＋Ser206Asp＋Thr207Asn＋Tyr208Asp＋Ala209SerGly196Asn＋Tyr203Ala＋Pro204Ser＋Ser206Asp＋Tyr208Asp＋Ala209Ser＋Asn212Ser＋Thr214GlnVal197Gly＋Asn198Ser＋Tyr203Asn＋Pro204Gly＋Gly205Ser＋Thr207Pro＋Asn212Asp＋Thr214AspAla194Gln＋Val197Asp＋Asn198Gln＋Tyr203His＋Ser210Glu＋Leu211Ala＋Gly213Ser＋Thr214AsnAla194Pro＋Val197Glu＋Val199His＋Gln200Ser＋Tyr203His＋Pro204Asn＋Tyr208Thr＋Thr214GluGly196Asn＋Asn198Asp＋Val199Gln＋Gln200Ser＋Thr202Ser＋Pro204Ser＋Leu211Cys＋Gly213GluAsn198Asp＋Tyr203Met＋Pro204Gly＋Thr207Asn＋Tyr208Gln＋Ala209Glu＋Asn212Ser＋Thr214SerAsn198Glu＋Val199Gly＋Thr202Gly＋Thr207Gln＋Tyr208Cys＋Ala209Asp＋Leu211Thr＋Asn212GlnAla194His＋Val197Asp＋Asn198Gln＋Gln200Glu＋Thr202Asn＋Tyr208Pro＋Leu211Ser＋Asn212SerPro195Gly＋Gln200Asp＋Tyr203Gly＋Gly205Asn＋Leu211Ser＋Asn212Glu＋Gly213Gln＋Thr214GlnAla194Asn＋Gly196Ser＋Val199Pro＋Gln200Glu＋Thr207Gly＋Tyr208Asn＋Ala209Thr＋Asn212GluAla194Thr＋Val199Asn＋Gln200Glu＋Tyr203Gln＋Pro204Gly＋Ala209His＋Asn212Glu＋Thr214SerAla194Asn＋Gly196Pro＋Val197Ser＋Asn198Gln＋Tyr203Pro＋Pro204Asp＋Tyr208Asn＋Ala209AspPro195Ser＋Val199Cys＋Gln200Ser＋Pro204Glu＋Gly205Ser＋Tyr208Ala＋Ala209Glu＋Asn212GlnAsn198Gln＋Val199His＋Thr202Gly＋Pro204Glu＋Gly205Pro＋Tyr208Ala＋Ala209Glu＋Asn212GlnGly196Ser＋Asn198Ser＋Val199Gly＋Gln200Ser＋Gly205Ser＋Tyr208Asp＋Leu211Asp＋Thr214SerPro195Gln＋Gly196Asn＋Val197Gly＋Gln200Asn＋Pro204Asn＋Thr207Gly＋Ala209Asp＋Asn212GluAla194Ser＋Gln200Ser＋Gly205Ser＋Ala209Asp＋Leu211Gly＋Asn212Asp＋Gly213Gln＋Thr214AsnAla194Gln＋Pro195Gln＋Gly196Pro＋Tyr203Leu＋Pro204Ser＋Ser206Asp＋Ala209Glu＋Leu211ProThr202Asn＋Tyr203Ala＋Pro204Ser＋Ser206Asp＋Tyr208Cys＋Ala209Glu＋Leu21lCys＋Asn212SerGly196Ser＋Val199His＋Gln200Asn＋Gly205Asn＋Ser206Glu＋Ala209Asp＋Leu211His＋Asn212SerAla194Asn＋Val197Ala＋Gln200Ser＋Ser206Glu＋Thr207Pro＋Ala209Glu＋Asn212Gln＋Gly213AsnPro195Gln＋Val197Met＋Val199Thr＋Gln200Ser＋Tyr203Ala＋Ala209Ser＋Ser210Glu＋Gly213GluAsn198Ser＋Val199Asn＋Ala209Pro＋Ser210Glu＋Leu211Gly＋Asn212Ser＋Gly213Asp＋Thr214GlyAla194Gly＋Asn198Gln＋Tyr203Ala＋Gly205Asp＋Tyr208Gln＋Ala209Asp＋Leu211Ala＋Asn212GlnAla194Gly＋Gly196Gln＋Thr202Gln＋Tyr203Thr＋Thr207Asp＋Tyr208Thr＋Ser210Glu＋Asn212SerAla194Pro＋Val197Thr＋Tyr203Pro＋Pro204Ser＋Thr207Asp＋Ser210Asp＋Leu211Pro＋Thr214SerPro195Asn＋Val199Ala＋Thr202Ser＋Gly205Gln＋Thr207Asp＋Ala209His＋Ser210Glu＋Gly213AsnPro195Gly＋Gly196Ser＋Val197Pro＋Pro204Gly＋Thr207Glu＋Tyr208Gln＋Ala209His＋Ser210AspAla194Asn＋Pro195Asn＋Asn198Ser＋Val199His＋Thr207Asp＋Tyr208Val＋Ala209Pro＋Ser210GluAla194Gly＋Gly196Ser＋Asn198Asp＋Thr207Asn＋Ala209Gln＋Leu21lPro＋Gly213Gln＋Thr214AspAla194Gly＋Pro195Ser＋Val197His＋Asn198Asp＋Gln200Asn＋Pro204Ser＋Asn212Ser＋Thr214AspVal199Asn＋Gln200Glu＋Pro204Gly＋Gly205Asn＋Tyr208Ser＋Leu211Ala＋Asn212Ser＋Gly213GluAla194Gln＋Pro195Ser＋Gly196Asn＋Asn198Gln＋Gln200Glu＋Tyr203Cys＋Leu21lSer＋Gly213GluAla194Ser＋Gly196Asn＋Val197Thr＋Asn198Gln＋Gln200Glu＋Thr207Asn＋Tyr208His＋Gly213AspAla194Gln＋Val197Gln＋Asn198Asp＋Gln200Asn＋Gly205Gln＋Tyr208Asp＋Leu211Gly＋Thr214SerAla194Ser＋Asn198Ser＋Thr202Pro＋Pro204Asp＋Gly205Gln＋Thr207Asn＋Ser210Asp＋Asn212SerPro195Gln＋Gly196Asn＋Pro204Glu＋Gly205Asn＋Thr207Asn＋Tyr208Ser＋Ala209Pro＋Ser210AspAla194Thr＋Gly196Pro＋Val199Pro＋Gln200Asn＋Pro204Glu＋Ala209Thr＋Ser210Asp＋Leu211AlaAla194Ser＋Gly196Gln＋Val199Thr＋Pro204Glu＋Tyr208Ile＋Ala209Gln＋Ser210Asp＋Thr214GlyAla194His＋Val197Ser＋Val199Ser＋Gln200Ser＋Pro204Glu＋Thr207Gly＋Ala209Pro＋Ser210AspAla194Gln＋Val197Ala＋Asn198Ser＋Pro204Asp＋Tyr208Cys＋Ala209Gly＋Ser210Asp＋Asn212SerGly196Ser＋Val197Thr＋Asn198Ser＋Gln200Glu＋Thr202Pro＋Pro204Glu＋Ala209Thr＋Gly213SerPro195Asn＋Val197Pro＋Val199Gly＋Gln200Asp＋Pro204Glu＋Thr207Gly＋Leu211Thr＋Gly213SerAla194Gln＋Val197Asn＋Gln200Ser＋Thr202Gln＋Thr207Glu＋Ala209Gly＋Leu211Glu＋Gly213AsnAla194Ser＋Gly196Ser＋Val197Ser＋Asn198Gln＋Gly205Gln＋Ser210Glu＋Asn212Ser＋Thr214AspGly196Ser＋Val199Cys＋Gln200Ser＋Thr207Pro＋Tyr208Pro＋Ala209Gly＋Ser210Asp＋Thr214AspVal197Cys＋Val199Gly＋Tyr203Gly＋Tyr208Met＋Ala209Ser＋Ser210Glu＋Leu211Ala＋Thr214GluAla194Pro＋Pro195Ser＋Gly196Ser＋Val199Asn＋Thr202Gly＋Ser210Asp＋Asn212Gln＋Thr214GluAla194Asn＋Gly196Asn＋Gln200Ser＋Pro204Asn＋Gly205Ser＋Ala209Ser＋Ser210Asp＋Thr214AspAla194His＋Val197Cys＋Tyr208Ile＋Ser210Glu＋Leu211Gly＋Asn212Ser＋Gly213Asn＋Thr214AspGly196Gln＋Gln200Asn＋Thr202Pro＋Tyr203Cys＋Gly205Gln＋Tyr208Ser＋Ser210Glu＋Thr214AspAsn198Gln＋Val199Pro＋Gln200Glu＋Thr202Pro＋Pro204Ser＋Ser206Glu＋Tyr208Gln＋Thr214AsnPro195Gly＋Gln200Asp＋Tyr203Val＋Ser206Asp＋Tyr208Thr＋Leu211Gly＋Asn212Gln＋Gly213AsnGly196Gln＋Gln200Glu＋Thr202Gln＋Tyr203Val＋Ser206Glu＋Tyr208Val＋Ala209Asn＋Leu211GlyPro195Gly＋Gln200Glu＋Thr202Pro＋Gly205Ser＋Ser206Glu＋Ala209Gln＋Leu211His＋Asn212GlnAsn198Gln＋Gln200Glu＋Pro204Gly＋Gly205Gln＋Ser206Asp＋Tyr208Met＋Asn212Ser＋Thr214AsnAla194Pro＋Pro195Gln＋Gly196Ser＋Val197His＋Gln200Glu＋Gly205Ser＋Ser206Glu＋Leu211SerAla194Gln＋Gln200Asp＋Tyr203Gln＋Pro204Asn＋Ser206Glu＋Tyr208Ile＋Ala209Gly＋Gly213AsnAla194Gln＋Pro195Gln＋Val197His＋Gln200Glu＋Pro204Ser＋Ser206Asp＋Tyr208Gln＋Ala209AsnPro195Gln＋Val197His＋Val199Asn＋Gln200Asp＋Thr202Pro＋Ser206Glu＋Leu211Thr＋Thr214ProPro195Gly＋Asn198Gln＋Val199Cys＋Gln200Asp＋Thr202Gly＋Ser206Glu＋Tyr208Cys＋Gly213SerAla194Gly＋Pro195Asn＋Gly196Asn＋Pro204Asp＋Thr207Ser＋Leu211Glu＋Asn212Gln＋Thr214Asn表31第6环状区-九突变的突变体Ala194Gly＋Pro195Ser＋Val197Cys＋Val199Asn＋Gln200Asp＋Tyr203Pro＋Pro204Ser＋Gly205Ser＋Leu211GlnAla194Pro＋Gln200Asn＋Thr202Asn＋Tyr203Asn＋Pro204Ser＋Tyr208Asn＋Ala209Asn＋Ser210Asp＋Thr214GlnPro195Gly＋Val199Ala＋Gln200Ser＋Thr202Gly＋Gly205Pro＋Ser206Glu＋Ala209His＋Leu211Ala＋Gly213AsnPro195Gln＋Gly196Gln＋Val197Thr＋Val199His＋Thr202Gln＋Tyr203Gly＋Pro204Gly＋Gly205Glu＋Gly213GlnPro195Gly＋Val197Gly＋Val199Asn＋Pro204Asn＋Ser206Asp＋Thr207Ser＋Tyr208Gln＋Ala209Gly＋Thr214AsnAla194Thr＋Val197Asn＋Val199Pro＋Gln200Asp＋Thr202Ser＋Tyr203Asn＋Thr207Asn＋Tyr208Leu＋Ala209GlyPro195Asn＋Gly196Asn＋Asn198Gln＋Tyr203Gln＋Thr207Ser＋Ala209Thr＋Leu211Ala＋Asn212Asp＋Thr214AsnAla194Gln＋Pro195Gly＋Gly196Pro＋Val197Gly＋Gln200Asp＋Thr202Ser＋Pro204Gly＋Tyr208Thr＋Thr214GlnGly196Asn＋Val199Gly＋Tyr203Ser＋Pro204Ser＋Tyr208Gln＋Ala209His＋Asn212Ser＋Gly213Asp＋Thr214GlnVal197His＋Gln200Ser＋Thr202Asn＋Pro204Ser＋Thr207Pro＋Tyr208Gly＋Ala209His＋Ser210Glu＋Leu211AlaPro195Ser＋Val197His＋Gln200Glu＋Thr202Gln＋Pro204Ser＋Tyr208Val＋Ala209His＋Leu211Thr＋Thr214ProPro195Gly＋Gly196Pro＋Val199Thr＋Thr202Pro＋Tyr203Ile＋Thr207Gly＋Ala209Pro＋Ser210Asp＋Asn212GlnAla194Gly＋Pro195Gln＋Val197Asp＋Tyr203Cys＋Tyr208Val＋Ala209Gln＋Leu211Gly＋Asn212Gln＋Gly213GlnGly196Asn＋Asn198Gln＋Thr202Gly＋Thr207Asp＋Tyr208Gly＋Ala209Ser＋Leu211Thr＋Asn212Ser＋Thr214ProAla194Pro＋Gly196Asn＋Val197Met＋Gln200Glu＋Tyr203Gln＋Gly205Asn＋Tyr208Asn＋Ala209Thr＋Thr214GlnAla194Pro＋Gly196Gln＋Val197Asn＋Gly205Ser＋Ser206Asp＋Thr207Asn＋Ala209Ser＋Leu211Cys＋Thr214ProGly196Gln＋Val197Thr＋Gln200Ser＋Tyr203Asn＋Pro204Asn＋Gly205Pro＋Tyr208Ser＋Ser210Glu＋Asn212SerAla194Thr＋Pro195Ser＋Gly196Pro＋Val197Cys＋Gln200Glu＋Pro204Ser＋Gly205Asn＋Ala209His＋Leu211ProPro195Gln＋Gly196Asn＋Asn198Gln＋Val199Gln＋Gln200Asp＋Pro204Ser＋Gly205Asn＋Thr207Ser＋Tyr208SerAla194His＋Val197Asn＋Tyr203Gly＋Pro204Gln＋Thr207Gly＋Ala209Gln＋Leu211His＋Asn212Gln＋Gly213GlnAla194Ser＋Pro195Gln＋Val197Met＋Val199His＋Thr207Pro＋Tyr208Ala＋Asn212Gln＋Gly213Glu＋Thr214AsnGly196Pro＋Val197His＋Asn198Gln＋Gln200Glu＋Tyr203Ser＋Gly205Pro＋Thr207Ser＋Leu211Thr＋Gly213AsnGly196Ser＋Val197Asn＋Val199Thr＋Thr202Gly＋Tyr203Asn＋Pro204Asn＋Ala209Gln＋Leu211Pro＋Thr214AspAla194Thr＋Pro195Ser＋Val197Asn＋Tyr203Met＋Tyr208Ala＋Ala209Ser＋Asn212Gln＋Gly213Asp＋Thr214AspAla194Asn＋Gly196Gln＋Val197Thr＋Val199Ala＋Gly205Asn＋Ala209Pro＋Leu211Asn＋Gly213Glu＋Thr214AspPro195Asn＋Val199Met＋Gln200Asn＋Thr202Asn＋Tyr203Cys＋Ala209Ser＋Leu211Gln＋Gly213Asp＋Thr214GluPro195Asn＋Val197Met＋Asn198Ser＋Val199Asn＋Gln200Asn＋Tyr208Gln＋Leu211His＋Gly213Glu＋Thr214AspVal197His＋Val199Ala＋Gln200Ser＋Pro204Ser＋Ala209Glu＋ Ser210Glu＋Asn212Gln＋Gly213Pro＋Thr214SerGly196Gln＋Val197Asn＋Asn198Gln＋Thr202Gly＋Gly205Gln＋Ala209Asp＋Ser210Glu＋Leu211Met＋Thr214GlyVal197Cys＋Asn198Ser＋Val199Gln＋Tyr203Gly＋Pro204Gln＋Ala209Glu＋Ser210Glu＋Gly213Pro＋Thr214GlyPro195Asn＋Val199His＋Gln200Ser＋Pro204Gln＋Gly205Gln＋Ala209Glu＋Ser210Asp＋Leu211Gln＋Asn212GlnVal197Asp＋Asn198Glu＋Val199Met＋Thr202Gln＋Pro204Ser＋Gly205Gln＋Thr207Gln＋Tyr208His＋Asn212SerAla194Thr＋Val197Glu＋Asn198Asp＋Val199His＋Tyr203Asn＋Tyr208His＋Ala209Ser＋Gly213Ser＋Thr214AsnVal197Cys＋Asn198Ser＋Val199Pro＋Gln200Ser＋Tyr203Ala＋Tyr208Asn＋Asn212Asp＋Gly213Glu＋Thr214SerGly196Asn＋Val197Met＋Asn198Ser＋Thr202Gly＋Pro204Gln＋Ala209Ser＋Leu211His＋Asn212Asp＋Gly213AspAla194Thr＋Pro195Ser＋Gln200Asn＋Thr202Asn＋Tyr203Gln＋Tyr208Ser＋Leu211Ala＋Asn212Asp＋Gly213AspPro195Gly＋Asn198Ser＋Val199Pro＋Gly205Asp＋Ser206Asp＋Thr207Pro＋Tyr208Ala＋Asn212Ser＋Gly213SerPro195Gln＋Asn198Ser＋Gln200Asn＋Thr202Ser＋Tyr203Met＋Gly205Asp＋Ser206Glu＋Thr207Pro＋Thr214SerGly196Ser＋Val199Thr＋Gln200Asn＋Tyr203Ala＋Gly205Asp＋Ser206Asp＋Tyr208Leu＋Ala209Thr＋Gly213ProAla194Pro＋Gly196Ser＋Val197Asn＋Val199His＋Gln200Asn＋Gly205Glu＋Ser206Glu＋Asn212Gln＋Thr214GlnAla194Thr＋Val199Ser＋Thr202Asn＋Gly205Glu＋Ser206Asp＋Ala209Pro＋Leu211Pro＋Asn212Gln＋Gly213GlnAsn198Ser＋Thr202Ser＋Tyr203Ser＋Pro204Gly＋Gly205Asn＋Ser206Asp＋Thr207Glu＋Ala209Pro＋Thr214SerPro195Asn＋Gly196Asn＋Val197Pro＋Val199Ser＋Thr202Ser＋Ser206Glu＋Thr207Glu＋Leu211Ile＋Gly213SerThr202Ser＋Tyr203Thr＋Thr207Asp＋Tyr208Glu＋Ala209Pro＋Leu211Val＋Asn212Ser＋Gly213Ser＋Thr214AsnVal199Pro＋Gln200Ser＋Thr202Ser＋Thr207Asp＋Tyr208Asp＋Ala209Gly＋Leu211Met＋Asn212Ser＋Thr214ProAla194Gln＋Pro195Gly＋Val197Ala＋Gln200Glu＋Tyr203Thr＋Gly205Ser＋Ala209Pro＋Ser210Asp＋Thr214AsnAla194Pro＋Pro195Ser＋Val199Met＋Gln200Glu＋Thr202Gly＋Pro204Gln＋Gly205Asn＋Tyr208Gln＋Ser210AspAla194Gln＋Val199Pro＋Gln200Asp＋Pro204Gly＋Gly205Gln＋Thr207Pro＋Tyr208Asn＋Ala209Gln＋Ser210AspAla194Pro＋Gly196Pro＋Asn198Gln＋Gln200Asp＋Pro204Gly＋Gly205Asn＋Tyr208Ile＋Ser210Asp＋Asn212SerPro195Ser＋Asn198Ser＋Val199Ala＋Gln200Glu＋Tyr203Thr＋Gly205Gln＋Tyr208Met＋Ser210Glu＋Gly213ProPro195Asn＋Asn198Ser＋Gln200Asp＋Thr202Gln＋Tyr203Ala＋Gly205Pro＋Ser210Asp＋Leu211Ser＋Gly213SerPro195Asn＋Gly196Asn＋Val197Ala＋Asn198Ser＋Gln200Asp＋Tyr203Gln＋Pro204Asn＋Ala209Ser＋Ser210AspPro195Ser＋Val197Pro＋Asn198Gln＋Val199Pro＋Gln200Glu＋Thr202Gln＋Thr207Asn＋Ala209His＋Ser210GluPro195Ser＋Gly196Asn＋Asn198Gln＋Gln200Glu＋Thr202Ser＋Pro204Gly＋Thr207Asn＋Ser210Glu＋Thr214AsnAla194Ser＋Pro195Ser＋Val197His＋Val199Thr＋Gln200Glu＋Gly205Gln＋Thr207Gly＋Ser210Glu＋Gly213ProPro195Asn＋Val197Asp＋Asn198Ser＋Val199Asn＋Thr207Pro＋Tyr208Ala＋Leu211Val＋Asn212Glu＋Gly213SerPro195Asn＋Gly196Asn＋Val197Gln＋Asn198Glu＋Val199Thr＋Tyr203Asn＋Thr207Pro＋Ser210Glu＋Gly213GlnVal197Cys＋Asn198Asp＋Val199Ser＋Tyr203Gln＋Gly205Ser＋Thr207Ser＋Tyr208Leu＋Ser210Asp＋Leu211GlyAla194Pro＋Gly196Gln＋Val197Gly＋Asn198Glu＋Gln200Ser＋Tyr203Thr＋Pro204Ser＋Ala209His＋Ser210GluGly196Ser＋Val197His＋Asn198Glu＋Thr202Pro＋Tyr203Ile＋ Thr207Asn＋Tyr208Val＋Ser210Glu＋Thr214SerGly196Asn＋Val197Thr＋Asn198Asp＋Val199Ala＋Pro204Asn＋Ser210Asp＋Leu211Val＋Gly213Gln＋Thr214ProGly196Ser＋Asn198Glu＋Thr202Asn＋Gly205Ser＋Thr207Gln＋Tyr208Ile＋Ser210Glu＋Leu211Ser＋Asn212SerAla194Asn＋Gly196Pro＋Asn198Asp＋Thr202Gly＋Tyr208Ser＋Ala209Pro＋Ser210Glu＋Leu211Ala＋Thr214AsnAla194His＋Asn198Glu＋Val199Gly＋Gln200Asn＋Gly205Asn＋Ser210Glu＋Leu211His＋Asn212Gln＋Thr214SerAla194Gly＋Asn198Glu＋Gln200Asn＋Thr202Pro＋Tyr203Cys＋Thr207Gln＋Tyr208Met＋Ser210Asp＋Asn212GlnPro195Gly＋Gly196Gln＋Asn198Glu＋Gln200Ser＋Pro204Gln＋Tyr208Leu＋Ala209Thr＋Ser210Glu＋Leu211IleAla194Pro＋Pro195Gln＋Val197His＋Asn198Glu＋Thr202Gln＋Gly205Pro＋Thr207Asn＋Tyr208Val＋Leu211AspPro195Gln＋Asn198Glu＋Thr202Ser＋Tyr203Gly＋Pro204Ser＋Thr207Gln＋Ala209Pro＋Leu211Asp＋Gly213AsnVal199Asn＋Gln200Asp＋Thr202Gln＋Tyr203Gly＋Pro204Asn＋Thr207Gln＋Tyr208Ala＋Ala209Glu＋Asn212SerGly196Pro＋Val197Thr＋Pro204Asp＋Ser206Glu＋Ala209Gln＋Leu211Gln＋Asn212Ser＋Gly213Ser＋Thr214ProGly196Pro＋Asn198Ser＋Val199Thr＋Pro204Glu＋Ser206Asp＋Thr207Gln＋Ala209Gly＋Asn212Ser＋Thr214GlyAla194Asn＋Val197Gly＋Gln200Ser＋Thr202Ser＋Pro204Asp＋Ser206Asp＋Tyr208Gln＋Ala209Gln＋Leu211GlyAla194Pro＋Thr202Gln＋Tyr203Asn＋Pro204Asp＋Ser206Glu＋Tyr208Gly＋Ala209Ser＋Gly213Pro＋Thr214ProAla194Asn＋Pro195Gln＋Val197Thr＋Asn198Ser＋Pro204Asp＋Gly205Ser＋Ser206Asp＋Asn212Ser＋Thr214AsnPro195Gly＋Gln200Ser＋Thr202Gln＋Tyr203Pro＋Pro204Asp＋Gly205Asn＋Ser206Glu＋Thr207Asn＋Leu211HisAla194His＋Gly196Asn＋Val197Met＋Asn198Ser＋Pro204Asp＋Ser206Asp＋Leu211Asn＋Asn212Ser＋Thr214GlnAsn198Gln＋Val199Cys＋Gln200Ser＋Thr202Gln＋Pro204Asp＋Gly205Gln＋Ser206Asp＋Tyr208Leu＋Thr214ProAla194His＋Pro195Ser＋Gly196Pro＋Asn198Gln＋Pro204Glu＋Ser206Asp＋Thr207Pro＋Tyr208His＋Ala209AsnAla194Ser＋Asn198Asp＋Gln200Ser＋Thr202Gly＋Thr207Pro＋Tyr208Asn＋Ala209Gln＋Leu211Ser＋Asn212GluGly196Asn＋Asn198Gln＋Gln200Asn＋Tyr203Thr＋Pro204Ser＋Gly205Asp＋Thr207Glu＋Ala209Ser＋Thr214AsnVal197His＋Asn198Gln＋Val199Met＋Gln200Ser＋Pro204Glu＋Gly205Ser＋Thr207Glu＋Tyr208Val＋Thr214GlyVal197Asn＋Asn198Asp＋Val199Ala＋Gln200Glu＋Tyr203Cys＋Pro204Gly＋Gly205Asn＋Thr207Gln＋Gly213AsnPro195Asn＋Gly196Gln＋Asn198Asp＋Val199Pro＋Gln200Glu＋Tyr203Val＋Gly205Asn＋Tyr208Cys＋Ala209ProPro195Gly＋Gly196Pro＋Val197Asp＋Thr202Ser＋Pro204Gln＋Thr207Gly＋Tyr208Ile＋Asn212Gln＋Gly213GluAla194Pro＋Vall97Glu＋Tyr203His＋Gly205Pro＋Thr207Gly＋Ala209Asn＋Leu211His＋Gly213Asp＋Thr214GlyPro195Gln＋Val197Glu＋Asn198Gln＋Val199Ala＋Pro204Ser＋Gly205Gln＋Ala209Ser＋Gly213Glu＋Thr214SerPro195Asn＋Val197Asp＋Asn198Gln＋Val199Ser＋Gln200Ser＋Thr207Ser＋Tyr208Ile＋Asn212Ser＋Gly213AspGly196Pro＋Val197Glu＋Asn198Gln＋Val199Asn＋Pro204Gly＋Thr207Pro＋Leu211His＋Gly213Asp＋Thr214AsnAla194Ser＋Asn198Gln＋Thr202Pro＋Tyr203Met＋Pro204Ser＋Thr207Gln＋Ala209Asp＋Leu211Glu＋Thr214GlnGly196Pro＋Val197Pro＋Thr202Asn＋Tyr203Cys＋Tyr208Gln＋Ser210Asp＋Asn212Asp＋Gly213Pro＋Thr214SerPro195Gln＋Tyr203Thr＋Gly205Gln＋Ala209Gln＋Ser210Asp＋Leu211Asn＋Asn212Glu＋Gly213Asn＋Thr214GlnAla194Gly＋Pro195Ser＋Val197His＋Thr202Gln＋Pro204Gly＋Ser210Asp＋Leu211His＋Asn212Glu＋Thr214GlyPro195Asn＋Gly196Asn＋Val197Asn＋Thr202Ser＋Tyr203Val＋Gly205Ser＋Ser210Asp＋Asn212Glu＋Gly213ProGln200Ser＋Thr202Gly＋Tyr203Leu＋Thr207Pro＋Tyr208Pro＋Ser210Asp＋Leu211Pro＋Asn212Glu＋Thr214GlyAla194Gln＋Gly196Ser＋Val197Cys＋Val199Met＋Tyr203Leu＋Ala209Thr＋Ser210Asp＋Asn212Glu＋Thr214SerVal197Met＋Val199Ala＋Gln200Asn＋Tyr203Thr＋Thr207Gln＋Tyr208Glu＋Ser210Glu＋Asn212Ser＋Thr214GlyGly196Ser＋Val197Ala＋Val199Ser＋Thr202Gly＋Tyr203Cys＋Gly205Asn＋Ser206Glu＋Tyr208Asp＋Leu211IleAla194Gly＋Pro195Ser＋Val199His＋Thr202Asn＋Gly205Gln＋Ser206Asp＋Tyr208Glu＋Asn212Ser＋Thr214ProGly196Pro＋Val199Gln＋Thr202Ser＋Gly205Pro＋Ser206Asp＋Thr207Gln＋Tyr208Glu＋Leu211His＋Asn212SerVal197His＋Asn198Ser＋Gly205Ser＋Thr207Pro＋Tyr208Val＋Leu211Val＋Asn212Glu＋Gly213Ser＋Thr214GluPro195Ser＋Gly196Pro＋Val199Thr＋Tyr203Ser＋Tyr208Gly＋Ala209Asn＋Asn212Glu＋Gly213Ser＋Thr214GluPro195Asn＋Asn198Gln＋Gln200Asn＋Thr202Asn＋Thr207Asn＋Tyr208Gln＋Asn212Glu＋Gly213Ser＋Thr214GluAla194Asn＋Val199Ser＋Thr202Gly＋Pro204Asn＋Gly205Ser＋Ala209Asn＋Leu211Asn＋Asn212Asp＋Thr214GluAla194Ser＋Gly196Gln＋Pro204Asp＋Gly205Pro＋Thr207Pro＋Tyr208Glu＋Leu211Ile＋Gly213Asn＋Thr214SerPro195Ser＋Val197Asp＋Asn198Gln＋Gln200Asn＋Tyr203Pro＋Gly205Asn＋Ser210Asp＋Asn212Ser＋Thr214AsnAla194His＋Gly196Ser＋Val197Asp＋Asn198Ser＋Gln200Asn＋Thr202Ser＋Gly205Gln＋Ser210Glu＋Asn212SerAla194Thr＋Val197Glu＋Thr202Gln＋Pro204Gln＋Tyr208Gly＋Ala209Thr＋Ser210Glu＋Asn212Ser＋Thr214GlnGly196Ser＋Val197Asp＋Val199Pro＋Tyr203Cys＋Pro204Ser＋Thr207Asn＋Tyr208Asn＋Ser210Asp＋Asn212SerAla194Pro＋Pro195Ser＋Gly196Gln＋Val197Asp＋Val199Met＋Gln200Asn＋Leu211Met＋Gly213Pro＋Thr214AspPro195Asn＋Gly196Gln＋Asn198Gln＋Val199Ala＋Gln200Ser＋Tyr203Pro＋Gly205Glu＋Thr207Asn＋Tyr208AspGly196Pro＋Asn198Ser＋Val199Asn＋Thr202Gly＋Pro204Gln＋Gly205Asp＋Thr207Gln＋Tyr208Asp＋Thr214AsnVal197Ser＋Asn198Glu＋Gln200Asn＋Tyr203Gln＋Pro204Asn＋Gly205Pro＋Thr207Gly＋Ala209Thr＋Gly213GluAla194Thr＋Gly196Gln＋Val197His＋Asn198Asp＋Val199Ser＋Tyr203Pro＋Tyr208Cys＋Gly213Asp＋Thr214ProAsn198Asp＋Gln200Ser＋Tyr203Gln＋Pro204Ser＋Gly205Asn＋Thr207Asn＋Ala209Glu＋Leu211Gln＋Asn212SerPro195Gly＋Gly196Asn＋Asn198Glu＋Thr202Gln＋Tyr203Ala＋Pro204Ser＋Thr207Pro＋Ala209Glu＋Gly213GlnAla194Pro＋Asn198Glu＋Val199Thr＋Pro204Gly＋Thr207Gly＋Tyr208Ile＋Ala209Asp＋Leu211Met＋Asn212SerAla194Ser＋Val197Glu＋Val199Thr＋Gln200Asp＋Tyr203Gly＋Gly205Ser＋Tyr208His＋Ala209Thr＋Asn212SerGly196Gln＋Val197Glu＋Val199Pro＋Gln200Asp＋Gly205Gln＋Thr207Ser＋Ala209Thr＋Leu211Pro＋Thr214GlyVal197Gly＋Gln200Glu＋Thr202Gly＋Tyr203Leu＋Gly205Gln＋Thr207Gly＋Tyr208Leu＋Ala209Asn＋Asn212AspPro195Ser＋Gly196Asn＋Asn198Ser＋Val199His＋Gln200Glu＋Tyr203Pro＋Ala209Ser＋Asn212Asp＋Thr214AsnAsn198Ser＋Val199Met＋Gln200Glu＋Thr202Ser＋Tyr203Ile＋Gly205Pro＋Ala209Pro＋Asn212Glu＋Gly213GlnGly196Pro＋Val197Ser＋Val199Pro＋Gln200Glu＋Ala209Gly＋Leu211Gln＋Asn212Asp＋Gly213Pro＋Thr214AsnAla194Gln＋Gly196Ser＋Val197Pro＋Thr202Pro＋Pro204Asp＋Tyr208Gly＋Ala209Glu＋Gly213Asn＋Thr214ProAla194Pro＋Val197Ser＋Val199His＋Gln200Asn＋Pro204Gly＋Ser206Glu＋Thr207Asn＋Tyr208Leu＋Ala209GluGly196Asn＋Thr202Asn＋Pro204Asn＋Gly205Pro＋Ser206Glu＋Tyr208Asn＋Ala209Asp＋Leu211Met＋Thr214GlnPro195Gly＋Asn198Gln＋Val199Ser＋Gln200Ser＋Tyr203Thr＋Gly205Pro＋Ser206Glu＋Ala209Glu＋Thr214AsnAla194Thr＋Val197Ala＋Thr202Ser＋Tyr203Pro＋Gly205Asn＋Ser206Glu＋Ala209Glu＋Leu211Gln＋Thr214AsnPro195Ser＋Val197Ala＋Thr202Asn＋Pro204Gly＋Ser206Asp＋Thr207Asn＋Ala209Glu＋Asn212Ser＋Thr214SerAla194Thr＋Val199Asn＋Thr202Pro＋Tyr203Ile＋Pro204Asn＋Ser206Asp＋Thr207Asn＋Ala209Glu＋Thr214AsnAla194Ser＋Pro195Asn＋Val197Gly＋Val199Gly＋Gln200Glu＋Thr202Gln＋Pro204Asn＋Thr207Asp＋Leu211GlnAla194Gly＋Val197His＋Gln200Ser＋Pro204Asn＋Thr207Ser＋Ser210Glu＋Leu211Gly＋Gly213Glu＋Thr214AsnAla194Gln＋Gly196Gln＋Val197Met＋Val199His＋Thr202Pro＋Gly205Asn＋Ala209Ser＋Ser210Glu＋Gly213GluPro195Asn＋Val197Thr＋Val199Ala＋Gln200Asn＋Tyr203Asn＋Thr207Gly＋Ser210Asp＋Asn212Gln＋Gly213GluAla194His＋Val199Pro＋Gln200Asn＋Thr202Pro＋Tyr203Asn＋Gly205Glu＋Ala209Glu＋Asn212Gln＋Thr214AsnGly196Pro＋Asn198Gln＋Gln200Asn＋Tyr203Gln＋Pro204Gln＋Gly205Asp＋Ala209Glu＋Asn212Gln＋Thr214GlnPro195Gln＋Asn198Gln＋Val199Ala＋Gln200Asn＋Thr207Glu＋Tyr208Leu＋Ala209Ser＋Ser210Asp＋Gly213ProPro195Gly＋Gly196Gln＋Val199Ser＋Gln200Asn＋Pro204Ser＋Gly205Ser＋Thr207Glu＋Ser210Glu＋Thr214ProGly196Gln＋Gln200Ser＋Thr202Gly＋Tyr203Ile＋Thr207Asp＋Tyr208Ile＋Ser210Asp＋Leu211Asn＋Asn212SerAla194His＋Gly196Gln＋Val197Gly＋Thr207Glu＋Tyr208Val＋Ala209Gln＋Ser210Asp＋Leu211His＋Gly213SerAla194Gln＋Asn198Asp＋Val199Ser＋Gln200Ser＋Tyr203Ile＋Gly205Asn＋Thr207Asn＋Asn212Gln＋Thr214AspVal197Pro＋Asn198Glu＋Val199Thr＋Gln200Ser＋Pro204Asn＋Gly205Pro＋Leu211Cys＋Asn212Ser＋Thr214AspAla194Ser＋Val199Thr＋Gln200Glu＋Tyr203Cys＋Tyr208Gly＋Ala209His＋Leu211Gly＋Asn212Ser＋Gly213GluGly196Gln＋Asn198Gln＋Val199Met＋Gln200Asp＋Pro204Gln＋Tyr208Pro＋Asn212Gln＋Gly213Asp＋Thr214GlnVal197Asp＋Val199Gln＋Gln200Ser＋Thr202Pro＋Thr207Gln＋Ala209Glu＋Leu211Gly＋Asn212Ser＋Gly213GlnAla194Ser＋Gly196Pro＋Val197Glu＋Asn198Ser＋Val199Met＋Pro204Gly＋Thr207Gln＋Ala209Glu＋Gly213GlnVal197Asp＋Asn198Ser＋Pro204Gln＋Gly205Gln＋Thr207Gly＋Tyr208Met＋Ala209Glu＋Asn212Gln＋Thr214GlnAla194Ser＋Val197Glu＋Asn198Gln＋Tyr203Asn＋Gly205Asn＋Ala209Glu＋Asn212Gln＋Gly213Ser＋Thr214ProPro195Gly＋Val197Ser＋Asn198Ser＋Tyr203Leu＋Pro204Glu＋Ser210Asp＋Leu211Ser＋Asn212Ser＋Thr214GlnAla194Asn＋Pro195Gln＋Asn198Gln＋Val199Asn＋Gln200Asn＋Pro204Asp＋Thr207Pro＋Ser210Glu＋Gly213GlnAla194Pro＋Val197Ser＋Asn198Ser＋Thr202Pro＋Tyr203Val＋Pro204Glu＋Ser210Asp＋Asn212Ser＋Thr214AsnAla194Gln＋Pro195Ser＋Gly196Pro＋Val199Ser＋Pro204Asp＋Tyr20SCys＋Ala209Thr＋Ser210Glu＋Gly213AsnPro195Ser＋Tyr203Pro＋Pro204Asp＋Gly205Pro＋Thr207Gln＋Ala209Gly＋Ser210Asp＋Leu211Asn＋Thr214AsnGly196Ser＋Asn198Ser＋Thr202Pro＋Pro204Asp＋Tyr208Val＋Ala209Gln＋Ser210Asp＋Leu211Asn＋Asn212GlnGly196Ser＋Val199Met＋Tyr203Ala＋Pro204Glu＋Gly205Ser＋Thr207Ser＋Tyr208Leu＋Ala209Gln＋Ser210AspPro195Gly＋Val197Pro＋Thr202Asn＋Pro204Glu＋Gly205Ser＋Thr207Asn＋Tyr208Pro＋Ser210Glu＋Asn212SerAla194Asn＋Gly196Ser＋Asn198Ser＋Val199Met＋Gln200Asn＋Thr202Gly＋Thr207Gln＋Ala209Asp＋Gly213AspAla194Gln＋Pro195Gln＋Thr202Ser＋Pro204Asn＋Ala209Thr＋Ser210Glu＋Leu211Asn＋Gly213Gln＋Thr214GluGly196Gln＋Val197Pro＋Asn198Gln＋Val199Gly＋Ala209Pro＋Ser210Asp＋Leu211Cys＋Asn212Ser＋Thr214GluAla194Asn＋Pro195Gly＋Asn198Gln＋Val199His＋Tyr203Pro＋Thr207Gly＋Tyr208Met＋Ser210Asp＋Thr214AspAla194Pro＋Pro195Ser＋Gly196Asn＋Thr202Gly＋Pro204Ser＋Ala209His＋Ser210Glu＋Leu211Ser＋Thr214AspAla194Thr＋Gly196Pro＋Asn198Ser＋Thr207Asn＋Tyr208Ile＋Ser210Asp＋Leu211Val＋Gly213Gln＋Thr214GluAla194Gly＋Pro195Ser＋Asn198Ser＋Thr202Ser＋Gly205Ser＋Ala209Gln＋Ser210Asp＋Asn212Ser＋Thr214AspAla194Ser＋Pro195Gly＋Gly196Ser＋Val199Ala＋Tyr203Gln＋Pro204Gln＋Thr207Ser＋Ser210Glu＋Thr214GluGly196Ser＋Gln200Asn＋Thr202Pro＋Tyr203Leu＋Thr207Gln＋Ser210Asp＋Leu211Ala＋Gly213Pro＋Thr214AspAla194Gln＋Asn198Ser＋Val199Thr＋Gln200Ser＋Tyr203Thr＋Ala209Pro＋Ser210Asp＋Leu211Asn＋Thr214GluPro195Gln＋Gly196Gln＋Tyr203Cys＋Pro204Ser＋Gly205Pro＋Tyr208His＋Ala209Thr＋Ser210Asp＋Thr214GluGly196Gln＋Asn198Ser＋Val199Thr＋Gln200Ser＋Thr202Ser＋Thr207Ser＋Ala209His＋Ser210Glu＋Thr214GluPro195Asn＋Gly196Pro＋Asn198Gln＋Tyr203Ala＋Thr207Ser＋Ala209Pro＋Ser210Asp＋Leu211Thr＋Thr214AspAla194Ser＋Pro195Asn＋Val199Pro＋Pro204Asn＋Gly205Gln＋Thr207Gln＋Ser210Glu＋Gly213Ser＋Thr214AspGly196Ser＋Val197Asn＋Thr202Gln＋Tyr203Ser＋Pro204Ser＋Gly205Ser＋Ser210Glu＋Leu21lCys＋Thr214AspVal197Asn＋Gln200Asp＋Tyr203Cys＋Pro204Asn＋Gly205Asn＋Ser206Asp＋Thr207Ser＋Tyr208Leu＋Asn212GlnPro195Asn＋Gln200Asp＋Thr202Gln＋Tyr203Gln＋Pro204Ser＋Ser206Glu＋Thr207Pro＋Asn212Ser＋Gly213GlnAla194Gln＋Pro195Gln＋Gly196Asn＋Val199Ser＋Gln200Asp＋Thr202Ser＋Gly205Ser＋Ser206Glu＋Ala209SerPro195Asn＋Gly196Pro＋Val197Cys＋Val199Met＋Gln200Asp＋Ser206Glu＋Thr207Ser＋Ala209Gln＋Leu211ValPro195Gly＋Val199Pro＋Gln200Asp＋Thr202Ser＋Tyr203Gly＋Pro204Gln＋Ser206Asp＋Leu211Met＋Asn212SerGly196Pro＋Val197Cys＋Asn198Gln＋Gln200Glu＋Thr202Asn＋Ser206Asp＋Ala209Asn＋Leu211Cys＋Gly213SerPro195Gln＋Asn198Ser＋Gln200Glu＋Thr202Asn＋Pro204Gly＋Ser206Asp＋Thr207Ser＋Tyr208Ser＋Asn212GlnGly196Asn＋Asn198Ser＋Val199Met＋Gln200Asp＋Thr202Ser＋Tyr203His＋Pro204Asn＋Gly205Gln＋Ser206GluAla194Thr＋Asn198Gln＋Val199Ala＋Gln200Asp＋Thr202Asn＋Ser206Glu＋Thr207Ser＋Tyr208His＋Gly213GlnAla194Gln＋Gly196Gln＋Asn198Gln＋Gln200Asp＋Thr202Asn＋Ser206Glu＋Thr207Ser＋Tyr208Thr＋Gly213Gln表32第6环状区-十突变的突变体Ala194Ser＋Gly196Gln＋Val197Ala＋Asn198Gln＋Thr202Pro＋Pro204Ser＋Gly205Ser＋Thr207Pro＋Leu211Val＋Asn212AspPro195Ser＋Val197Gln＋Val199His＋Thr202Asn＋Tyr203Pro＋Gly205Gln＋Ala209Thr＋Ser210Glu＋Asn212Ser＋Thr214GlnAla194Gly＋Asn198Ser＋Val199Ala＋Thr202Gln＋Tyr203Leu＋Pro204Gly＋Gly205Pro＋Ala209His＋Gly213Ser＋Thr214AspAla194His＋Gly196Gln＋Thr202Gly＋Tyr203Val＋Gly205Asn＋Thr207Gln＋Tyr208Cys＋Ala209Ser＋Gly213Gln＋Thr214AsnAla194Thr＋Pro195Ser＋Gly196Asn＋Asn198Glu＋Gln200Asn＋Thr202Gly＋Pro204Ser＋Asn212Gln＋Gly213Pro＋Thr214GlyAla194Ser＋Pro195Asn＋Gly196Asn＋Val199Thr＋Thr202Gln＋Pro204Asp＋Thr207Asn＋Tyr208Ser＋Leu211Asn＋Asn212GlnPro195Asn＋Gly196Ser＋Val197Gly＋Asn198Ser＋Thr202Gly＋Gly205Pro＋Thr207Pro＋Tyr208Asp＋Ala209Ser＋Thr214AsnAla194His＋Pro195Gln＋Gly196Asn＋Asn198Gln＋Val199His＋Pro204Ser＋Gly205Asn＋Thr207Ser＋Tyr208Ser＋Thr214SerAla194Pro＋Pro195Asn＋Val197Asn＋Val199Cys＋Gln200Ser＋Thr202Pro＋Pro204Ser＋Ser206Glu＋Tyr208His＋Leu211IleAla194Thr＋Val197Met＋Asn198Asp＋Val199Gly＋Gln200Ser＋Tyr203Gln＋Pro204Asn＋Thr207Gln＋Ala209Pro＋Leu211AlaAla194Pro＋Pro195Gly＋Val197Asn＋Asn198Ser＋Val199His＋Tyr208His＋Ala209Asp＋Leu211Thr＋Asn212Gln＋Thr214GlnPro195Ser＋Val199Met＋Thr202Ser＋Tyr203Ile＋Gly205Gln＋Ser206Asp＋Tyr208His＋Leu211Pro＋Asn212Gln＋Gly213ProAla194Thr＋Val197Ser＋Val199Asn＋Thr202Gln＋Tyr203Met＋Gly205Pro＋Ser206Glu＋Tyr208Ile＋Ala209Asn＋Gly213SerAla194Thr＋Val197Gln＋Asn198Gln＋Val199Gly＋Thr202Ser＋Pro204Ser＋Gly205Pro＋Thr207Asn＋Ala209Pro＋Thr214AsnAla194His＋Pro195Asn＋Gly196Asn＋Val199Thr＋Gln200Asn＋Thr202Asn＋Tyr203Ala＋Gly205Asp＋Thr207Gly＋Thr214ProAla194Ser＋Pro195Gly＋Gly196Ser＋Val197Pro＋Asn198Gln＋Val199Thr＋Thr202Pro＋Tyr203Met＋Pro204Gln＋Gly205GlnAla194Thr＋Pro195Gln＋Val197Asp＋Gln200Ser＋Thr202Pro＋Pro204Ser＋Gly205Gln＋Thr207Gly＋Tyr208His＋Ala209GlyAla194Pro＋Pro195Ser＋Asn198Ser＋Val199Gln＋Gln200Asp＋Thr202Gln＋Tyr203Asn＋Pro204Gly＋Leu211Val＋Thr214ProPro195Ser＋Gly196Gln＋Val199Pro＋Gln200Asn＋Tyr203Gln＋Thr207Asp＋Ala209His＋Leu211His＋Asn212Gln＋Thr214GlyGly196Pro＋Asn198Ser＋Val199Gly＋Gln200Ser＋Thr202Asn＋Tyr203Ser＋Tyr208Leu＋Ala209His＋Leu211Asp＋Asn212AspGly196Asn＋Val199Ala＋Gln200Ser＋Thr202Gln＋Tyr203His＋Pro204Asn＋Gly205Pro＋Asn212Gln＋Gly213Glu＋Thr214GluGly196Pro＋Val199Ser＋Gln200Ser＋Thr202Pro＋Pro204Gln＋Ala209Glu＋Ser210Glu＋Asn212Ser＋Gly213Ser＋Thr214SerVal197Thr＋Asn198Gln＋Val199Gln＋Pro204Gly＋Thr207Pro＋Ala209Asp＋Ser210Glu＋Asn212Ser＋Gly213Ser＋Thr214SerAla194Pro＋Pro195Gln＋Val197Ala＋Val199Gln＋Gln200Ser＋Pro204Gln＋Gly205Pro＋Tyr208Met＋Ala209Asp＋Ser210GluPro195Gly＋Val197Gly＋Asn198Gln＋Thr202Gly＋Tyr203Ala＋Gly205Pro＋Ala209Glu＋Ser210Glu＋Leu211Ser＋Gly213GlnPro195Ser＋Gly196Asn＋Asn198Gln＋Gln200Ser＋Thr202Gly＋Pro204Gln＋Tyr208Thr＋Leu211Cys＋Asn212Glu＋Gly213GluGly196Gln＋Val197Ser＋Gln200Ser＋Thr202Pro＋Pro204Asp＋Gly205Asp＋Tyr208Gln＋Ala209Pro＋Asn212Ser＋Thr214GlnAla194Pro＋Pro195Gln＋Gly196Gln＋Pro204Asp＋Gly205Glu＋Tyr208Ala＋Ala209Ser＋Leu211Asn＋Gly213Asn＋Thr214SerAla194Thr＋Val197Cys＋Val199Met＋Gln200Ser＋Gly205Pro＋Tyr208Thr＋Ser210Glu＋Leu211Asp＋Asn212Ser＋Gly213ProAla194Gly＋Val197His＋Thr202Ser＋Tyr203Asn＋Thr207Pro＋Tyr208Cys＋Ser210Glu＋Leu211Glu＋Asn212Gln＋Gly213ProVal197His＋Asn198Ser＋Gln200Ser＋Thr202Asn＋Gly205Ser＋Tyr208Gln＋Ser210Glu＋Leu211Glu＋Asn212Ser＋Thr214ProAla194Thr＋Gly196Asn＋Asn198Gln＋Gln200Asn＋Thr202Pro＋Tyr203Thr＋Gly205Asn＋Ser210Glu＋Leu211Glu＋Asn212SerAla194Gly＋Gly196Gln＋Asn198Ser＋Val199Cys＋Pro204Asn＋Gly205Asp＋Ser206Asp＋Thr207Gln＋Ala209Gly＋Gly213AsnPro195Gln＋Val197Met＋Val199Ala＋Gln200Ser＋Gly205Asp＋Ser206Asp＋Thr207Asn＋Tyr208Gln＋Leu211Pro＋Asn212SerAla194His＋Pro195Gln＋Asn198Gln＋Thr202Pro＋Gly205Glu＋Ser206Asp＋Thr207Pro＋Leu211Thr＋Gly213Ser＋Thr214GlnGly196Pro＋Asn198Gln＋Val199Gly＋Gln200Ser＋Thr202Asn＋Pro204Asn＋Gly205Glu＋Ser206Asp＋Thr207Gly＋Thr214GlnPro195Gln＋Val197His＋Asn198Gln＋Thr202Gly＋Gly205Asp＋Ser206Glu＋Thr207Ser＋Tyr208Ala＋Asn212Gln＋Gly213AsnAla194Asn＋Gly196Asn＋Val197Ser＋Asn198Ser＋Val199Met＋Thr202Asn＋Tyr203Ala＋Gly205Asp＋Ser206Asp＋Thr214AsnAla194Thr＋Pro195Asn＋Gly196Gln＋Val199Thr＋Gln200Asn＋Thr202Ser＋Pro204Asn＋Gly205Asp＋Ser206Glu＋Thr207AsnPro195Ser＋Gly196Ser＋Val197Pro＋Asn198Gln＋Gln200Asn＋Tyr203Leu＋Gly205Asp＋Ser206Glu＋Asn212Ser＋Thr214ProPro195Ser＋Gly196Pro＋Val197Ser＋Gln200Ser＋Thr202Pro＋Tyr203Val＋Pro204Gly＋Gly205Glu＋Ser206Glu＋Tyr208MetAla194Gly＋Gly196Gln＋Gln200Ser＋Thr202Ser＋Pro204Asn＋Ser206Asp＋Thr207Asp＋Ala209Gln＋Leu211Met＋Thr214ProGly196Asn＋Val197Ser＋Val199Ala＋Tyr203Ile＋Ser206Asp＋Thr207Glu＋Ala209Ser＋Leu211Ala＋Asn212Ser＋Thr214AsnAla194Ser＋Pro204Asn＋Gly205Ser＋Ser206Asp＋Thr207Glu＋Tyr208Met＋Ala209Asn＋Leu211Ile＋Asn212Gln＋Gly213AsnAla194Gln＋Asn198Gln＋Val199Asn＋Thr202Gln＋Tyr203Ile＋Gly205Gln＋Ser206Asp＋Thr207Glu＋Tyr208Ser＋Gly213SerGly196Gln＋Asn198Gln＋Thr202Gln＋Pro204Ser＋Gly205Ser＋Ser206Glu＋Thr207Asp＋Leu211Ile＋Gly213Pro＋Thr214ProPro195Asn＋Gly196Gln＋Val197Gln＋Thr202Asn＋Gly205Ser＋Ser206Glu＋Thr207Glu＋Tyr208Ile＋Gly213Gln＋Thr214AsnAsn198Ser＋Thr202Ser＋Tyr203Ile＋Gly205Asn＋Thr207Asn＋Tyr208Asp＋Ala209Glu＋Leu211Ile＋Gly213Ser＋Thr214SerVal199His＋Gln200Glu＋Tyr203Met＋Pro204Gly＋Thr207Pro＋Ala209Thr＋Ser210Glu＋Leu211Asn＋Asn212Ser＋Gly213ProAla194His＋Gly196Ser＋Asn198Ser＋Gln200Glu＋Thr202Gln＋Tyr203Gln＋Gly205Pro＋Thr207Gly＋Ser210Glu＋Gly213AsnGly196Pro＋Val199Asn＋Gln200Glu＋Pro204Gly＋Tyr208His＋Ser210Asp＋Leu211Gln＋Asn212Ser＋Gly213Gln＋Thr214AsnPro195Gly＋Gly196Pro＋Gln200Asp＋Tyr203Asn＋Pro204Asn＋Gly205Asn＋Thr207Asn＋Tyr208Ile＋Ser210Asp＋Gly213GlnAla194Gln＋Pro195Gln＋Gly196Gln＋Val197Gln＋Gln200Asp＋Thr207Gly＋Ala209Asn＋Ser210Asp＋Leu211Gly＋Gly213AsnVal197Thr＋Val199Ser＋Gln200Glu＋Tyr203Asn＋Pro204Gly＋Thr207Asn＋Tyr208Gly＋Ser210Glu＋Asn212Ser＋Thr214AsnGly196Gln＋Val197Gln＋Val199Asn＋Gln200Glu＋Tyr203Asn＋Thr207Gln＋Ala209Gly＋Ser210Glu＋Asn212Gln＋Gly213GlnPro195Asn＋Val197Met＋Val199Cys＋Gln200Asp＋Thr202Asn＋Thr207Ser＋Tyr208Gly＋Ala209Gln＋Ser210Glu＋Leu211AsnAla194Asn＋Val199Gln＋Gln200Asp＋Tyr203Gly＋Pro204Ser＋Gly205Pro＋Tyr208Ile＋Ser210Glu＋Leu211Asn＋Gly213SerAla194His＋Pro195Gln＋Val197Glu＋Asn198Ser＋Val199Ser＋Tyr203His＋Thr207Pro＋Leu211Gln＋Asn212Glu＋Gly213SerGly196Ser＋Val197Glu＋Asn198Gln＋Val199Cys＋Tyr203Pro＋Pro204Gly＋Ala209Gln＋Asn212Glu＋Gly213Gln＋Thr214AsnGly196Ser＋Val197Gly＋Asn198Asp＋Gln200Ser＋Tyr203Pro＋Tyr208Ala＋Ala209Gly＋Ser210Glu＋Leu211Ile＋Thr214AsnGly196Pro＋Asn198Asp＋Val199Gln＋Gln200Asn＋Pro204Gln＋Gly205Pro＋Thr207Gly＋Ser210Asp＋Asn212Gln＋Gly213SerPro195Asn＋Gly196Ser＋Val197Gln＋Asn198Glu＋Gln200Ser＋Pro204Gly＋Tyr208Gln＋Ala209Asn＋Ser210Glu＋Gly213ProAsn198Asp＋Val199Met＋Gln200Asn＋Thr202Gln＋Pro204Asn＋Gly205Gln＋Tyr208Ala＋Ala209His＋Ser210Glu＋Thr214ProAla194Gln＋Val197Ala＋Asn198Glu＋Gln200Asn＋Thr202Gly＋Pro204Gln＋Gly205Gln＋Tyr208Thr＋Ser210Asp＋Leu211ValVal197Ser＋Asn198Asp＋Val199Asn＋Gln200Ser＋Pro204Ser＋Thr207Pro＋Tyr208Leu＋Ser210Glu＋Asn212Gln＋Thr214SerAla194Thr＋Gly196Asn＋Val197Ser＋Asn198Glu＋Val199Asn＋Gln200Ser＋Thr202Ser＋Thr207Ser＋Ser210Glu＋Gly213SerGly196Pro＋Val197Met＋Asn198Asp＋Val199Gly＋Gln200Asn＋Tyr203Pro＋Gly205Asn＋Tyr208Val＋Ser210Asp＋Leu211HisGly196Gln＋Asn198Asp＋Gln200Asn＋Tyr203Asn＋Gly205Pro＋Thr207Ser＋Tyr208Gln＋Ala209Pro＋Leu211Glu＋Asn212GlnPro195Asn＋Val197Pro＋Asn198Glu＋Val199Cys＋Tyr203Asn＋Thr207Gly＋Tyr208Gln＋Ala209Ser＋Leu211Glu＋Gly213AsnAla194Gln＋Gly196Gln＋Val199Met＋Gln200Asp＋Tyr203Cys＋Thr207Ser＋Ala209Asp＋Asn212Ser＋Gly213Gln＋Thr214GlnPro195Asn＋Gly196Pro＋Asn198Ser＋Gln200Glu＋Thr202Ser＋Thr207Gln＋Tyr208Ala＋Ala209Glu＋Leu211Ala＋Asn212GlnPro195Gly＋Gly196Asn＋Val199Gln＋Thr202Gly＋Pro204Glu＋Gly205Gln＋Ser206Glu＋Thr207Ser＋Leu211Gln＋Asn212GlnPro195Gly＋Gln200Asn＋Tyr203Leu＋Pro204Glu＋Ser206Asp＋Tyr208Gln＋Ala209Gln＋Asn212Gln＋Gly213Pro＋Thr214GlnAla194His＋Pro195Ser＋Val199Thr＋Thr202Gly＋Tyr203Cys＋Pro204Glu＋Ser206Glu＋Leu211His＋Asn212Ser＋Gly213ProPro195Gln＋Val197Gln＋Asn198Gln＋Gln200Ser＋Pro204Glu＋Gly205Ser＋Ser206Asp＋Ala209Gly＋Asn212Ser＋Thr214GlnAla194Thr＋Asn198Asp＋Val199Ala＋Gln200Asn＋Gly205Pro＋Tyr208Ser＋Ala209Gly＋Leu211Val＋Asn212Glu＋Gly213GlnAla194Gly＋Pro195Gln＋Val199Cys＋Tyr203Leu＋Pro204Ser＋Gly205Glu＋Thr207Asp＋Ala209Gln＋Asn212Gln＋Thr214GlyPro195Gln＋Val197Gln＋Gln200Ser＋Thr202Gln＋Pro204Glu＋Gly205Ser＋Thr207Asp＋Leu211His＋Asn212Ser＋Thr214SerAla194Asn＋Pro195Ser＋Val197Ser＋Asn198Gln＋Thr202Asn＋Pro204Asp＋Thr207Glu＋Leu211His＋Asn212Ser＋Gly213SerAla194Thr＋Pro195Ser＋Val197Gln＋Asn198Glu＋Val199Met＋Gln200Asp＋Pro204Gly＋Gly205Ser＋Ala209Asn＋Thr214AsnPro195Gln＋Gly196Pro＋Val197Met＋Asn198Asp＋Val199Ala＋Gln200Glu＋Thr202Pro＋Tyr203Ile＋Tyr208Cys＋Leu211ThrAla194Gln＋Gly196Asn＋Asn198Glu＋Val199Ala＋Gln200Asp＋Thr202Ser＋Tyr203Cys＋Thr207Asn＋Tyr208Met＋Gly213AsnAla194Thr＋Val197Pro＋Val199His＋Gln200Asp＋Thr202Gln＋Tyr203Met＋Gly205Ser＋Thr207Asn＋Tyr208Asp＋Asn212SerPro195Asn＋Val197Asp＋Asn198Gln＋Val199Gly＋Gln200Ser＋Thr207Ser＋Tyr208Ile＋Ala209Asn＋Asn212Ser＋Gly213AspPro195Asn＋Val197Pro＋Asn198Gln＋Gln200Glu＋Gly205Gln＋Tyr208Gly＋Ala209Gly＋Leu211Glu＋Asn212Ser＋Thr214SerAla194Thr＋Gly196Ser＋Asn198Ser＋Gln200Asp＋Thr202Gly＋Tyr203Met＋Gly205Gln＋Tyr208Ala＋Leu211Asp＋Thr214AsnPro195Ser＋Asn198Ser＋Val199Thr＋Thr202Asn＋Gly205Asn＋Thr207Ser＋Ala209Gly＋Leu211Glu＋Asn212Gln＋Gly213AspPro195Gln＋Gly196Gln＋Asn198Ser＋Thr202Pro＋Pro204Asn＋Thr207Asp＋Tyr208Gly＋Ala209Asp＋Leu211Ile＋Asn212SerAla194Gly＋Pro195Gly＋Asn198Ser＋Gln200Asn＋Thr202Asn＋Tyr203Ala＋Tyr208Val＋Ser210Glu＋Asn212Asp＋Thr214GlyAla194Gln＋Pro195Gln＋Asn198Gln＋Gln200Asn＋Thr202Gly＋Tyr203Leu＋Gly205Gln＋Ala209Gln＋Ser210Glu＋Asn212GluPro195Gln＋Val197Gly＋Val199Gly＋Thr202Asn＋Gly205Pro＋Thr207Asn＋Ala209Asn＋Ser210Glu＋Leu211Met＋Asn212GluAla194Thr＋Pro195Gly＋Gly196Pro＋Asn198Ser＋Thr207Pro＋Tyr208Cys＋Ala209Ser＋Ser210Glu＋Leu211Ser＋Asn212AspAla194His＋Pro195Ser＋Gly196Gln＋Val197Ala＋Val199Asn＋Tyr203Gln＋Thr207Asn＋Tyr208His＋Ser210Asp＋Asn212GluAla194Gln＋Pro195Asn＋Gly196Ser＋Val197Met＋Pro204Ser＋Gly205Pro＋Thr207Ser＋Tyr208Ala＋Ser210Glu＋Asn212GluAla194Ser＋Val199Thr＋Thr202Ser＋Tyr203Ala＋Pro204Asn＋Gly205Pro＋Ser210Asp＋Leu211Ala＋Asn212Asp＋Thr214ProPro195Asn＋Thr202Gln＋Pro204Ser＋Thr207Gly＋Tyr208Glu＋Ala209Asn＋Ser210Glu＋Leu211Asn＋Asn212Gln＋Gly213AsnGly196Asn＋Val197Thr＋Val199Thr＋Pro204Ser＋Thr207Gln＋Tyr208Asp＋Ser210Asp＋Leu211Thr＋Asn212Gln＋Thr214AsnAla194Asn＋Pro195Asn＋Val199Cys＋Gln200Asn＋Pro204Gln＋Thr207Ser＋Tyr208Glu＋Ser210Asp＋Leu211Asn＋Asn212GlnAla194Thr＋Gly196Pro＋Val199His＋Gln200Ser＋Gly205Asn＋Ser206Glu＋Tyr208Glu＋Ala209Thr＋Leu211Met＋Thr214SerAla194Ser＋Pro195Gln＋Asn198Gln＋Val199His＋Pro204Asn＋Gly205Gln＋Ser206Glu＋Tyr208Asp＋Ala209Thr＋Gly213AsnGly196Gln＋Val197Met＋Val199Ser＋Gln200Ser＋Thr202Asn＋Pro204Asn＋Tyr208Thr＋Ala209Gln＋Asn212Glu＋Thr214AspVal197Ala＋Asn198Ser＋Gln200Ser＋Thr202Gly＋Pro204Gln＋Thr207Gln＋Tyr208Gln＋Ala209His＋Asn212Glu＋Thr214AspAla194Pro＋Gly196Ser＋Val197Asp＋Val199Asn＋Thr202Asn＋Tyr203Val＋Thr207Gln＋Ala209Asn＋Ser210Glu＋Asn212GlnPro195Ser＋Val197Glu＋Asn198Gln＋Val199Gln＋Tyr203Leu＋Tyr208Val＋Ala209Asn＋Asn212Gln＋Gly213Pro＋Thr214GluPro195Gln＋Gly196Pro＋Val197Asn＋Asn198Glu＋Gln200Ser＋Pro204Asn＋Gly205Asn＋Leu211Cys＋Asn212Gln＋Gly213AspPro195Gln＋Gly196Pro＋Val197Thr＋Asn198Asp＋Thr202Pro＋Tyr203Gln＋Thr207Gly＋Ala209Glu＋Gly213Ser＋Thr214SerAla194Ser＋Pro195Gln＋Val197Pro＋Asn198Asp＋Val199Ala＋Tyr203Thr＋Pro204Gln＋Ala209Asp＋Leu211Val＋Asn212SerPro195Gly＋Val197Glu＋Asn198Gln＋Gln200Asp＋Tyr203His＋Pro204Ser＋Thr207Gly＋Tyr208His＋Gly213Pro＋Thr214GlyPro195Gly＋Gly196Pro＋Val199Thr＋Gln200Glu＋Tyr203Met＋Pro204Gln＋Gly205Pro＋Tyr208Ser＋Asn212Asp＋Gly213ProGly196Pro＋Val197Ser＋Val199Pro＋Gln200Glu＋Thr207Gly＋Ala209Gly＋Leu211Gln＋Asn212Asp＋Gly213Pro＋Thr214AsnAla194His＋Pro195Gln＋Gly196Asn＋Val197Ser＋Gln200Glu＋Tyr203Ala＋Pro204Asn＋Gly205Ser＋Thr207Pro＋Asn212GluAla194Ser＋Pro195Asn＋Gly196Pro＋Asn198Ser＋Val199Thr＋Tyr208Ile＋Leu211Glu＋Asn212Gln＋Gly213Ser＋Thr214GluAla194Pro＋Pro195Asn＋Asn198Ser＋Val199Met＋Tyr203Met＋Gly205Pro＋Thr207Pro＋Tyr208Ser＋Leu211Glu＋Thr214GluAla194His＋Pro195Gln＋Gly196Gln＋Val199Pro＋Tyr203His＋Thr207Asn＋Ala209Asp＋Leu211Ala＋Asn212Glu＋Thr214AsnAla194His＋Gly196Gln＋Thr202Ser＋Tyr203Gly＋ Pro204Gly＋Gly205Ser＋Ser206Asp＋Thr207Gln＋Ala209Asp＋Thr214ProPro195Gln＋Thr202Ser＋Pro204Gln＋Gly205Pro＋Ser206Asp＋Tyr208Met＋Ala209Glu＋Asn212Gln＋Gly213Pro＋Thr214SerGly196Gln＋Val197Pro＋Thr202Asn＋Tyr203Gly＋Gly205Pro＋Ser206Glu＋Thr207Ser＋Ala209Glu＋Leu211Gln＋Asn212SerAla194Asn＋Pro195Ser＋Gly196Pro＋Val199Ala＋Thr202Gln＋Pro204Gln＋Ser206Asp＋Ala209Glu＋Leu211His＋Asn212SerGly196Ser＋Asn198Ser＋Val199Ser＋Gln200Asn＋Tyr203Asn＋Gly205Gln＋Ser206Glu＋Ala209Glu＋Leu211Ser＋Gly213GlnAla194Thr＋Val197Cys＋Val199Asn＋Pro204Gly＋Ser206Asp＋Thr207Gly＋Tyr208Gly＋Ala209Glu＋Leu211Ile＋Thr214GlnAla194Gly＋Pro195Ser＋Gln200Ser＋Thr202Gly＋Tyr203Leu＋Pro204Gly＋Gly205Pro＋Ser206Asp＋Ala209Asp＋Thr214GlyPro195Ser＋Gly196Pro＋Val199His＋Gln200Asp＋Thr202Pro＋Gly205Pro＋Thr207Asp＋Ala209Thr＋Asn212Gln＋Gly213GlnAla194Gln＋Gly196Asn＋Asn198Gln＋Val199Pro＋Gln200Asp＋Thr202Gly＋Tyr203Thr＋Gly205Pro＋Thr207Asp＋Ala209GlnPro195Asn＋Val197Thr＋Val199Ala＋Gln200Asn＋Tyr203Asn＋Gly205Pro＋Thr207Gly＋Ser210Asp＋Asn212Gln＋Gly213GluAla194Asn＋Pro195Gln＋Val197Asn＋Val199Asn＋Pro204Gly＋Thr207Pro＋Ser210Asp＋Asn212Ser＋Gly213Glu＋Thr214ProAla194Gln＋Pro195Asn＋Val197Gly＋Tyr203Met＋Pro204Ser＋Tyr208Gln＋Ala209Asn＋Ser210Glu＋Asn212Ser＋Gly213GluAla194His＋Gly196Ser＋Tyr203Asn＋Pro204Gly＋Gly205Pro＋Ala209Ser＋Ser210Glu＋Leu211Pro＋Asn212Ser＋Gly213AspGly196Pro＋Gln200Asn＋Thr202Asn＋Tyr203His＋Gly205Asp＋Tyr208Ile＋Ala209Glu＋Leu211Met＋Gly213Pro＋Thr214ProVal197His＋Val199Asn＋Gln200Asn＋Thr202Ser＋Tyr203Ser＋Gly205Glu＋Tyr208Ser＋Ala209Asp＋Gly213Pro＋Thr214ProAla194Asn＋Val197Pro＋Gln200Ser＋Thr202Gln＋Tyr203Met＋Gly205Gln＋Thr207Asp＋Ser210Asp＋Leu211Ser＋Asn212SerAla194Thr＋Asn198Ser＋Tyr203Asn＋Gly205Pro＋Thr207Asp＋Ala209Gln＋Ser210Glu＋Leu211Val＋Asn212Ser＋Thr214SerAsn198Gln＋Val199Pro＋Gln200Asn＋Thr202Gln＋Tyr203Asn＋Thr207Asp＋Ala209Pro＋Ser210Glu＋Asn212Ser＋Gly213AsnAla194Ser＋Pro195Asn＋Asn198Gln＋Val199Ala＋Gln200Asp＋Tyr203His＋Ala209Thr＋Leu211Ser＋Gly213Asp＋Thr214SerAla194Pro＋Pro195Asn＋Gly196Pro＋Val197Ser＋Gln200Asp＋Tyr208Cys＋Leu211Gln＋Asn212Ser＋Gly213Asp＋Thr214GlyAla194Gln＋Pro195Ser＋Gly196Gln＋Asn198Asp＋Gln200Ser＋Thr202Ser＋Tyr208Asp＋Ala209Thr＋Leu211His＋Thr214AsnAla194Asn＋Pro195Gln＋Val197Thr＋Asn198Asp＋Pro204Asn＋Thr207Ser＋Tyr208Asp＋Leu211Ile＋Gly213Gln＋Thr214GlnAla194Gln＋Pro195Asn＋Val199Asn＋Tyr203Ser＋Pro204Asp＋Thr207Asn＋Ser210Asp＋Leu21lGly＋Asn212Ser＋Gly213SerAla194Asn＋Pro195Asn＋Thr202Gly＋Pro204Asp＋Tyr208Ile＋Ala209Gly＋Ser210Asp＋Leu211Val＋Asn212Gln＋Thr214SerAla194Asn＋Gly196Ser＋Asn198Ser＋Val199Pro＋Tyr203His＋Pro204Glu＋Gly205Gln＋Tyr208Met＋Ala209Thr＋Ser210GluAla194Thr＋Asn198Ser＋Val199Thr＋Thr202Asn＋Pro204Glu＋Gly205Asn＋Thr207Gln＋Ala209His＋Ser210Glu＋Thr214GlyAla194His＋Pro195Gln＋Asn198Ser＋Val199Gly＋Tyr203Gln＋Pro204Asp＋Thr207Gly＋Ser210Glu＋Asn212Gln＋Gly213SerPro195Asn＋Tyr203Ser＋Pro204Glu＋Gly205Pro＋Tyr208His＋Ala209Gly＋Ser210Glu＋Leu211Pro＋Gly213Pro＋Thr214AsnGly196Asn＋Asn198Ser＋Gln200Ser＋Thr202Gly＋Pro204Asp＋Gly205Asn＋Ala209Thr＋Ser210Glu＋Gly213Pro＋Thr214SerGln200Ser＋Thr202Gln＋Tyr203Met＋Pro204Asp＋Gly205Pro＋Tyr208Cys＋Ala209Gly＋Ser210Asp＋Leu211Thr＋Gly213GlnAla194Pro＋Gly196Asn＋Gln200Ser＋Tyr203Ser＋Pro204Glu＋Thr207Gly＋Ala209Asn＋Ser210Asp＋Leu211Val＋Thr214SerVal199His＋Gln200Ser＋Thr202Ser＋Pro204Asp＋Gly205Asn＋Tyr208Pro＋Ser210Asp＋Leu211Pro＋Gly213Pro＋Thr214ProAla194Ser＋Pro195Asn＋Gly196Ser＋Val197Ser＋Asn198Gin＋Val199Ser＋Pro204Glu＋Tyr208Leu＋Ser210Asp＋Leu211SerAla194His＋Gly196Pro＋Val199Gly＋Gln200Asp＋Pro204Glu＋Tyr208Gln＋Ala209Gly＋Leu211Ser＋Asn212Ser＋Thr214AsnAla194Gln＋Val197Pro＋Val199Asn＋Gln200Asp＋Thr202Gln＋Pro204Asp＋Gly205Asn＋Tyr208His＋Ala209Asn＋Leu211IlePro195Ser＋Val197Met＋Asn198Ser＋Gln200Asp＋Thr202Gly＋Tyr203Gln＋Pro204Glu＋Thr207Pro＋Asn212Ser＋Thr214GlyPro195Ser＋Val199Met＋Gln200Ser＋Pro204Gln＋Thr207Ser＋Ala209Glu＋Leu211Ala＋Asn212Gln＋Gly213Glu＋Thr214GlyPro195Gln＋Gly196Pro＋Asn198Ser＋Val199Gly＋Tyr203Asn＋Gly205Asn＋Thr207Pro＋Ala209Glu＋Leu211Val＋Gly213AspAla194Asn＋Pro195Gln＋Asn198Ser＋Val199Ser＋Gly205Asn＋Thr207Glu＋Ala209Ser＋Leu211Glu＋Asn212Gln＋Thr214SerAla194Ser＋Asn198Ser＋Gln200Ser＋Thr202Gly＋Tyr203Leu＋Thr207Gly＋Ala209His＋Ser210Glu＋Asn212Ser＋Thr214AspGly196Ser＋Gln200Asn＋Thr202Pro＋Tyr203Leu＋Pro204A＋Thr207Gln＋Ser210Asp＋Leu211Ala＋Gly213Pro＋Thr214AspGly196Ser＋Asn198Gln＋Tyr203Gly＋Pro204Gln＋Gly205Pro＋Thr207Pro＋Tyr208His＋Ser210Glu＋Gly213Gln＋Thr214AspVal197Thr＋Val199Thr＋Gln200Ser＋Thr202Ser＋Gly205Asn＋Thr207Pro＋Tyr208Cys＋Ser210Glu＋Gly213Ser＋Thr214AspGly196Gln＋Val197Thr＋Gln200Asp＋Thr202Asn＋Tyr203His＋Ser206Glu＋Thr207Ser＋Tyr208Pro＋Leu211Pro＋Asn212GlnVal197Met＋Val199Gln＋Gln200Glu＋Thr202Gln＋Pro204Gln＋Gly205Asn＋Ser206Asp＋Thr207Gln＋Leu211Val＋Thr214SerAla194Thr＋Pro195Gly＋Gly196Asn＋Val197Ser＋Gln200Glu＋Thr202Pro＋Gly205Pro＋Ser206Glu＋Leu211Met＋Asn212SerProl95Ser＋Gly196Gln＋Val199Cys＋Gln200Glu＋Ser206Glu＋Tyr208Thr＋Leu211Gln＋Asn212Gln＋Gly213Pro＋Thr214AsnGly196Gln＋Val197Gly＋Asn198Gln＋Gln200Glu＋Gly205Asn＋Ser206Glu＋Thr207Pro＋Ala209Asn＋Asn212Gln＋Gly213SerPro195Gln＋Val197Gly＋Gln200Glu＋Thr202Gln＋Tyr203Val＋Gly205Gln＋Ser206Asp＋Tyr208Met＋Ala209Pro＋Thr214GlyAla194Ser＋Pro195Asn＋Val197Thr＋Val199Thr＋Gln200Glu＋Thr202Pro＋Tyr203Ser＋Gly205Pro＋Ser206Glu＋Thr214GlyPro195Ser＋Gln200Glu＋Thr202Gln＋Tyr203Gly＋Gly205Pro＋Ser206Asp＋Thr207Gly＋Ala209His＋Leu211His＋Thr214GlyAla194Asn＋Pro195Gly＋Gly196Asn＋Val197Cys＋Val199Asn＋Gln200Glu＋Tyr203Pro＋Pro204Gln＋Ser206Asp＋Leu211AlaAla194Thr＋Pro195Asn＋Gly196Gln＋Val197Asn＋Gln200Glu＋Thr202Ser＋Ser206Glu＋Ala209His＋Leu211Ala＋Thr214SerVal199Gly＋Gln200Glu＋Thr202Gln＋Pro204Asn＋Gly205Pro＋Ser206Asp＋Thr207Ser＋Ala209Gly＋Leu211Cys＋Asn212SerAla194Thr＋Val197Ser＋Val199Gln＋Gln200Asp＋Thr202Gln＋Pro204Gln＋Ser206Glu＋Ala209Gly＋Leu211His＋Gly213GlnAla194Gly＋Gly196Asn＋Asn198Gln＋Gln200Glu＋Gly205Pro＋Ser206Asp＋Ala209His＋Leu211Pro＋Gly213Gln＋Thr214GlyGly196Ser＋Val199Ser＋Thr202Ser＋Pro204Asp＋Thr207Asn＋Tyr208Leu＋Leu211Glu＋Asn212Gln＋Gly213Ser＋Thr214ProAla194Pro＋Pro195Asn＋Val197Pro＋Thr202Pro＋Tyr203Thr＋Thr207Gln＋Tyr208Glu＋Asn212Glu＋Gly213Ser＋Thr214SerGly196Asn＋Gln200Ser＋Gly205Pro＋Thr207Gln＋Tyr208Asp＋Ala209Ser＋Leu211Ser＋Asn212Glu＋Gly213Pro＋Thr214ProAla194Gly＋Val199Pro＋Gln200Asp＋Thr202Pro＋Tyr203Cys＋Pro204Gly＋Ala209Gln＋Leu211Cys＋Asn212Gln＋Thr214AspGly196Ser＋Val197Gly＋Gln200Asp＋Tyr203Cys＋Pro204Ser＋Thr207Gln＋Leu211Ala＋Asn212Gln＋Gly213Ser＋Thr214AspAla194Pro＋Val197Thr＋Thr202Gln＋Tyr203Met＋Pro204Asn＋Ser206Asp＋Tyr208His＋Ala209His＋Ser210Asp＋Thr214AsnGly196Asn＋Val197Cys＋Thr202Asn＋Tyr203Met＋Gly205Pro＋Ser206Asp＋Thr207Gly＋Ala209Pro＋Ser210Asp＋Gly213GlnAla194Thr＋Val197Ser＋Val199His＋Thr202Gln＋Tyr203Ser＋Ser206Asp＋Thr207Ser＋Ser210Asp＋Gly213Pro＋Thr214ProAla194Thr＋Pro195Ser＋Gly196Asn＋Val197Pro＋Asn198Gln＋Thr202Gln＋Ser206Glu＋Ser210Glu＋Asn212Ser＋Thr214GlyPro195Asn＋Gly196Ser＋Val197Ser＋Asn198Gln＋Thr202Asn＋Tyr203Met＋Ser206Glu＋Ser210Asp＋Asn212Gln＋Thr214Gln表33多个环状区的双突变的突变体Leu 94Gly＋Gln200GluGln 57Ser＋Asn 60SerVal 93Gln＋Gly213AspTyr102Cys＋Thr207GlySer154Glu＋Asn198GlnLeu124Ile＋Asn198GlnAla209Gly＋Ser210GluGln 57Asp＋Leu 94GlyLeu 94Ala＋Ser154GluSer101Asp＋Leu211ThrAla131Glu＋Gly196GlnSer128Glu＋Pro204AsnSer103Asp＋Pro129AsnGly157Asn＋Thr207SerGly 98Glu＋Gly155GlnGln200Asp＋Ala209ProAsn 60Glu＋Asn198GlnLeu 94Ile＋Ser 99AspLeu124Ile＋Ser210AspSer126Glu＋Thr202SerVal 93Ala＋Pro127AspGly157Pro＋Ser210AspVal197Gly＋Thr207AspGln185Glu＋Ala194ThrSer 97Glu＋Tyr203AsnGly100Pro＋Thr207AspSer210Asp＋Leu211MetTyr161Asn＋Thr207AspLeu124Ser＋Pro127AsnGly 61Ser＋Thr207ProGly 98Glu＋Gln200SerGln200Asn＋Asn212GluAsp 58Glu＋Ile105CysGly 61Pro＋Ala209ThrLeu124Asp＋Pro204AsnGln 57Glu＋Asn198SerGln185Ser＋Asn212AspPhe183His＋Val197GluAsn 60Glu＋Gln200SerGly 59Asn＋Gly 61GluGly155Asp＋Ile159CysGly 63Gln＋Leu211GlyThr 64Pro＋Tyr161AspLeu 94Gln＋Pro127GlyThr207Gly＋Gly213GluGln185Asp＋Tyr203SerGly 63Asn＋Ala209AsnTyr208Met＋Gly213AsnSer210Glu＋Leu211AsnTyr102Ala＋Pro129AsnGly125Glu＋Leu211GlySer103Asp＋Thr207SerSer128Glu＋Leu211HisGly155Gln＋Leu211HisGln 57Asn＋Gln200AspSer103Asp＋Gly157GlnGly125Ser＋Thr207GlyIle105Pro＋Val197GluPro204Asp＋Ala209GlnTyrl61Asp＋Thr207AsnAsn198Ser＋Ser210AspGly 95Ser＋Gln200AspGly 59Gln＋Ala156GluGly 98Gln＋Tyrl02CysAla181His＋Thr207GluSer154Glu＋Pro204SerGly205Asn＋Thr207GlnSer210Glu＋Leu211ProLeu 94Gln＋Ser210GluAla209Asp＋Leu211ProAsn 60Glu＋Leu211HisAla 96Asn＋Thr214GlyAsn 60Glu＋Ala194GlySer103Asp＋Ala181HisGly125Glu＋Ala209AsnSer158Asp＋Asn198SerGly1555er＋Asn1985erSer104Asp＋Gly152AsnSer182Asp＋Leu211CysVal 93Ser＋Gly125GluAsn198Glu＋Leu211SerGly100Ser＋Ser103AspAla181Gln＋Gly213GlnThr 64Gly＋Ser126AspIle159Cys＋Gln200AspThr207Glu＋Leu211AsnSer104Glu＋Pro204GlySer130Asp＋Pro204AsnPro129Gln＋Gln185AsnAsn 60Glu＋Gly157SerGly125Pro＋Val197MetAsn 60Glu＋Ile105CysPro127Gln＋Val197ThrSer101Asp＋Leu211AlaGln 57Ser＋Ser 99GluPhe183Pro＋Asn198GluAsn198Glu＋Thr202ProVal197Gln＋Leu211HisGly 61Ser＋Gln200SerGly 61Asp＋Asn198GlnGly 61Glu＋Phe183SerThr207Asn＋Asn212GluTyr203Thr＋Tyr208GlyAsn198Ser＋Leu211CysAsn153Asp＋Leu211SerGln 57Asn＋Gly213ProThr 64Ser＋Ser210AspPro204Asp＋Leu211IleGln 57Asp＋Gly 61ProGly155Gln＋Ser210AspAla156Gly＋Thr207AsnGly 61Gln＋Ala181ThrIle159Cys＋Asn198GlnPro129Gly＋Leu211GluAsn198Glu＋Thr207SerGly 98Ser＋Ser128AspAla131Asn＋Val193SerGln185Ser＋Gln200AsnSer210Asp＋Leu211HisGly125Glu＋Ala156GlnSer 97Asp＋Gly125GlnAsn 60Gln＋Gly155ProSer126Glu＋Thr207SerPhe183Met＋Thr207ProSer154Glu＋Phe183IleVal 93Gln＋Ser210AspVal 93Gln＋Tyr208HisAsn 60Ser＋Asn198GluGly152Gln＋Gln200AspPro127Asn＋Gly152AsnGly155Gln＋Thr214AsnAla 96Ser＋Tyr208HisTyr161Thr＋Ser210AspGly100Pro＋Pro204GlnGln 57Asp＋Ala194AsnAsn 60Gln＋Thr202AsnGly157Pro＋Gly213AsnVal 93Thr＋Leu211AlaSer103Glu＋Leu211ThrSer101Asp＋Leu124IleLeu 94Glu＋Pro204GlnGln200Glu＋Ala209HisSer101Glu＋Gly152GlnAsn198Asp＋Gly205ProGly 61Ser＋Gln200AsnSer158Asp＋Val197AsnGly 61Glu＋Phe183IleAsp 58Glu＋Tyr203SerGly213Ser＋Thr214GlyGln 57Asp＋Asn198GlnTyr102Leu＋Ser158GluAla 96Gln＋Leu211AsnVal 93Asp＋Thr202AsnLeu 94Ser＋Asn198AspGly196Ser＋Asn212AspGly155Asn＋Gly157AspVal 93Asn＋Ser130AspLeu 94Ile＋Ser210AspVal197Ala＋Ser210AspSer104Asp＋Gly205ProAsn153Asp＋Ala181GlyGln200Asn＋Leu211GluLeu 94Asn＋Ser210GluGln185Asp＋Val197GlnTyr102Ser＋Thr207AspGly 61Glu＋Gly 95AsnGly 61Asp＋Pro129GlnSer 99Asp＋Thr207SerSer126Glu＋Gly152GlnVal197His＋Ser206GluGly 61Asn＋Gly155GluGly155Asn＋Asn198GluAla209Asn＋Ser210GluSer128Asp＋Thr207GlyAla209Thr＋Ser210AspGln185Asn＋Gly213ProSer126Glu＋Leu211GlySer 97Asp＋Gly205SerLeu 94Thr＋Leu124ThrAsn 60Gln＋Leu211GlyPro129Asp＋Gly155ProAsp 58Glu＋Gly157SerThr202Pro＋Thr207GluAla156Ser＋Tyr208AspGly 98Glu＋Tyr161GlnIle105Cys＋Gln200AsnGln200Glu＋Pro204GlyGly 98Pro＋Ala131AsnGly 59Asp＋Thr207GlnTyr102Gln＋Ala209GlnGly 98Glu＋Ile105HisSer101Asp＋Pro129GlnPro127Asp＋Gly213AsnSer104Glu＋Asn198GlnGln 57Ser＋Gln200AsnVal197Cys＋Gln200SerVal 93Ala＋Ala156GluGln200Asn＋Gly205GluLeu 94Ala＋Thr207AsnAla131Thr＋Gln200SerLeu 94Glu＋Tyr161ThrGln200Asn＋Ser210AspGln 57Asp＋Thr207AsnSer 99Glu＋Ala156ProLeu124Ile＋Asn198SerTyr161Gly＋Thr207GlnIle105Cys＋Thr214AspIle159His＋Ser206AspGly 63Gln＋Gly125AsnAsn198Asp＋Gly205SerSer104Glu＋Leu124GlnAla209Pro＋Ser210GluTyr161Val＋Asn198AspPro129Ser＋Gln200AspSer 97Asp＋Tyr203SerSer103Asp＋Phe183ThrTyr161Gly＋Val197AlaAsn198Gln＋Pro204GluLeu211Gly＋Asn212GluSer206Glu＋Leu211ThrGly152Pro＋Val197SerIle105Ala＋Ala156GlnGly 63Asn＋Tyr102ProSer182Glu＋Val197AsnThr 64Glu＋Ala 96GlnAsn 60Gln＋Ser206GluAsn 60Gln＋Ser128AspSer101Asp＋Ile159ProGly100Ser＋Thr207SerGly 63Asn＋Gly205SerAla181Glu＋Thr207AsnGly125Ser＋Tyr161SerGln200Glu＋Leu211AlaSer101Asp＋Tyr102LeuSer126Glu＋Thr207ProGly 59Asp＋Tyr102ThrGly100Gln＋Tyr102MetGly 59Ser＋Asn198AspAsn 60Ser＋Leu 94CysPro127Glu＋Gly205ProGln 57Glu＋Ala156AsnAla131Asn＋Thr207AspAla131Asp＋Thr202SerLeu124His＋Ala131AsnGly152Glu＋Leu211HisSerl01Glu＋Ala181ThrAsp 58Glu＋Gly205SerGly 61Asn＋Gln200AspPro129Gly＋Val197AspThr 64Gln＋Asn198SerVal 93Ala＋Ser154GluSer130Glu＋Tyr161ValPhe183Asp＋Leu211ThrSer206Glu＋Asn212GlnGly 63Ser＋Ala131GluPro127Asn＋Gln200AspSer130Asp＋Phe183ThrAla181Thr＋Pro195SerGly 95Ser＋Ala156GluTyr161Gln＋Tyr208IleGln 57Asp＋Leu211CysAsn153Gln＋Gly213AsnVal 93Thr＋Leu124SerSer101Glu＋Asn212GlnGly157Ser＋Asn198GluGly152Ser＋Ser182GluTyr161Gln＋Gly213SerPro195Gly＋Gln200AsnAsn 60Asp＋Leu211ProPro195Ser＋Tyr208GluGln200Asp＋Leu211AsnLeu124Val＋Thr207ProThr 64Pro＋Ser126GluThr 64Ser＋Tyr203ValSer 97Glu＋Asn198SerGly152Gln＋Leu211ProAsn198Asp＋Leu211AsnVal 93Glu＋Gly125GlnGly 98Asn＋Gln200AspGln200Asn＋Thr207AspAsn198Ser＋Gln200GluTyr102Leu＋Asn198GlnLeu124Cys＋Asn153SerAsn198Asp＋Leu211ProSer126Glu＋Tyr208MetAla 96Asn＋Gly100SerSer 99Glu＋Leu211CysSer 97Asp＋Leu211AlaSer182Glu＋Leu211HisIle159Cys＋Leu211GluGly152Asn＋Ser210AspVal197Ala＋Gln200GluAsn 60Ser＋Ser158GluSer104Glu＋Tyr208AsnPro204Asp＋Leu211GlyAsp 58Glu＋Val197GlySer182Glu＋Tyr208SerSer103Asp＋Ala194SerAla156Gln＋Thr207AspAsn198Ser＋Thr207SerGln 57Asp＋Thr207SerGly152Glu＋Thr207GlyAla131Ser＋Ile159AsnPro127Asn＋Gly152GluGln200Asn＋Gly213GluLeu124Asp＋Leu211AlaGly 98Glu＋Gly125AsnIle159Gly＋Thr207AsnPro127Gln＋Asn153AspSer103Glu＋Phe183TyrSer160Glu＋Pro204SerGly157Ser＋Tyr161HisAla156Pro＋Ser158GluGly100Glu＋Pro127AsnGln 57Asn＋Asn 60GluGly125Ser＋Ser210GluGly125Pro＋Ser128AspGly125Glu＋Val197GlyVal193Met＋Asn198SerSer160Glu＋Gly213AsnSer206Glu＋Ala209HisSer 97Glu＋Gly213SerGly100Asp＋Leu21lGlyPro129Gln＋Thr207GlyGly 95Gln＋Ser210AspGln185Ser＋Pro204GluPro129Glu＋Ala156SerSer126Asp＋Asn212SerGly 59Ser＋Ile159AlaAla 96Asn＋Ser 99GluSer104Asp＋Ile159ValGly 63Asn＋Ser210GluPhe183Pro＋Ser210GluAla 96Asp＋Tyr208IleGly 98Asp＋Thr207GlnVal 93Gln＋Gly125GluIle159Glu＋Thr207GlnLeu 94Ile＋Leu211ValGly157Asp＋Leu211HisGly 61Asp＋Leu211CysGln 57Asn＋Gly157AsnGly100Glu＋Val197SerGln200Glu＋Pro204AsnAla131Gly＋Leu211HisSer206Asp＋Leu211AsnAsn153Asp＋Ile159LeuGly152Ser＋Asn198GluLeu 94Met＋Ser130GluGln 57Glu＋Asn198GlnSer182Glu＋Leu211ThrVal199Ala＋Ser210GluThr207Gly＋Ser210GluThr 64Gly＋Gly 98AspGly 61Gln＋Ser 99AspGly155Glu＋Leu211SerLeu124Ser＋Thr207AspVal 93Ser＋Asn198GlnSer 99Asp＋Gly125AsnGln 57Asn＋Asn198GluSer 99Glu＋Ile159Met表34多个环状区的三突变的突变体Gln 57Ser＋Leu 94Gly＋Gln200GluAsn 60Ser＋Val 93Gln＋Gly213AspTyr102Cys＋Asn198Gln＋Thr207GlyLeu124Ile＋Ser154Glu＋Asn198GlnLeu 94Gly＋Ala209Gly＋Ser210GluGly155Asp＋Ala209Gln＋Asn212GlnPro129Asn＋Gly157Asn＋Thr207SerAsn198Gln＋Gln200Asp＋Ala209ProVal 93Ala＋Pro127Asp＋Thr202SerTyr161Asn＋Thr207Asp＋Leu211MetGly 61Ser＋Pro127Asn＋Thr207ProGly 61Pro＋Leu124Asp＋Pro204AsnGln 57Glu＋Gln185Ser＋Asn198SerGly 59Asn＋Asn 60Glu＋Gln200SerGly 63Gln＋Gln200Asp＋Leu211GlyTyr203Ser＋Thr207Gly＋Gly213GluGly 63Asn＋Gln185Asp＋Ala209AsnPro129Asn＋Ser210Glu＋Leu211AsnTyr102Ala＋Gly125Glu＋Leu211GlyGln 57Asn＋Gly155Gln＋Leu211HisIle105Pro＋Gly125Ser＋Thr207GlyAsn198Ser＋Thr207Asn＋Ser210AspGly 59Gln＋Gly 95Ser＋Ala156GluTyr102Cys＋Ser210Glu＋Leu211AsnGly205Asn＋Thr207Gln＋Ser210GluLeu 94Gln＋Ala209Asp＋Leu211ProAsn 60Glu＋Leu211His＋Thr214GlyAsn 60Glu＋Ala181His＋Ala194GlySer104Asp＋Gly155Ser＋Asn198SerGly152Asn＋Ser182Asp＋Leu211CysGly100Ser＋Ser103Asp＋Leu211SerThr 64Gly＋Ser126Asp＋Ile159CysSer130Asp＋Ala181Asn＋Pro204GlyPro129Gln＋Gly157Ser＋Gln185AsnAsn 60Glu＋Gly125Pro＋Val197MetIle105Cys＋Pro127Gln＋Val197ThrGln 57Ser＋Ser 99Glu＋Phe183ProGly 61Ser＋Val197Gln＋Leu211HisGly 61Asp＋Asn198Gln＋Gln200SerTyr203Thr＋Thr207Asn＋Tyr208GlyAsn153Asp＋Asn198Ser＋Leu211SerGln 57Asn＋Ser210Asp＋Gly213ProThr 64Ser＋Pro204Asp＋Leu211IleSer101Asp＋Ala156Gly＋Thr207AsnIle159Cys＋Ala181Thr＋Asn198GlnGly 98Ser＋Asn198Glu＋Thr207SerSer128Asp＋Ala131Asn＋Val193SerGln185Ser＋Gln200Asn＋Leu211HisSer126Glu＋Phe183Met＋Thr207SerSer154Glu＋Phe183Ile＋Thr207ProVal 93Gln＋Tyr208His＋Ser210AspVal 93His＋Pro127Asn＋Gly152GlnGly155Gln＋Gly213Glu＋Thr214AsnGly100Pro＋Tyr161Thr＋Ser210AspGln 57Asp＋Ala194Asn＋Pro204GlnGly157Pro＋Thr202Asn＋Gly213AsnThr 64Gln＋Val 93Thr＋Leu211AlaGly 61Ser＋Gln200Asn＋Gly205ProSer158Asp＋Phe183Ile＋Val197AsnAsn198Asp＋Gly213Ser＋Thr214GlyGly152Ser＋Gly155Ser＋Ser206AspGly152Gln＋Gln185Ser＋Val197GlnLeu 94Ile＋Val197Ala＋Ser210AspAla181Gly＋Gln200Asn＋Leu211GluLeu 94Asn＋Val197Gln＋Ser210GluGly 61Asp＋Pro129Gln＋Thr207SerGln 57Asn＋Gly 61Asn＋Gly155GluGly 61Pro＋Ser128Asp＋Ala209ThrThr207Gly＋Ala209Thr＋Ser210AspSer128Asp＋Gln185Asn＋Gly213ProLeu 94Thr＋Leu124Thr＋Gly205SerAsn 60Gln＋Gly155Pro＋Leu211GlyAsp 58Glu＋Gly1575er＋Thr202ProGly 98Glu＋Ile105Cys＋Gln200AsnGly 98Pro＋Gln200Glu＋Pro204GlyGly 59Asp＋Ala131Asn＋Thr207GlnGly 98Glu＋Tyr102Gln＋Ala209GlnSer101Asp＋Ile105His＋Pro129GlnPro127Asp＋Asn198Gln＋Gly213AsnIle159Gly＋Val197Gly＋Asn198GlnGln 57Ser＋Gly155Glu＋Gln200AsnVal197Cys＋Asn198Glu＋Gln200SerLeu 94Ala＋Gln200Asn＋Gly205GluAla131Thr＋Gln200Ser＋Thr207AsnLeu124Ile＋Ala156Pro＋Asn198SerVal 93Glu＋Tyr161Gly＋Thr207GlnGly 63Gln＋Gly125Asn＋Ala209GluTyr161Gly＋Val197Ala＋Asn198GlnSer158Asp＋Asn198Ser＋Leu211GlyIle105Ala＋Gly152Pro＋Val1975erGly 63Asn＋Tyr102Pro＋Ala156GlnAla 96Gln＋Ser182Glu＋Val197AsnAsn 60Gln＋Ser101Asp＋Ile159ProGly100Ser＋Gly205Ser＋Thr207SerTyr161Ser＋Ala181Glu＋Thr207AsnGly125Ser＋Gln200Glu＋Leu211AlaGly 59Asp＋Tyr102Thr＋Thr207ProGly 59Ser＋Gly100Gln＋Tyr102MetAsn 60Ser＋Leu 94Cys＋Asn198AspPro127Glu＋Ala156Asn＋Gly205ProAsp 58Glu＋Ala181Thr＋Gly205SerThr 64Gln＋Val 93Ala＋Asn198SerPhe183Asp＋Leu211Thr＋Asn212GlnPro127Asn＋Ser130Asp＋Phe183ThrTyr161Gln＋Tyr208Ile＋Leu211CysLeu124Ser＋Asn153Gln＋Gly213AsnVal 93Thr＋Ser101Glu＋Asn212GlnTyr161Gln＋Gln200Asn＋Gly213SerGln200Asp＋Thr207Pro＋Leu211AsnThr 64Pro＋Leu124Val＋Ser126GluThr 64Ser＋Ser 97Glu＋Asn198SerGly152Gln＋Ser210Asp＋Leu211ProVal 93Pro＋Asn198Ser＋Gln200GluThr 64Gln＋Tyr102Leu＋Asn153SerAla 96Asn＋Gly100Ser＋Tyr208MetIle159Cys＋Ser182Glu＋Leu211HisAsn 60Ser＋Ser158Glu＋Tyr208AsnSer103Asp＋Ala194Ser＋Tyr208SerAla156Gln＋Asn198Ser＋Thr207AspAla131Ser＋Gly152Glu＋Ile159AsnPro127Asn＋Gln200Asn＋Gly213GluGly 98Glu＋Gly125Asn＋Leu211AlaIle159Gly＋Thr207Asn＋Ser210GluSer103Glu＋Phe183Tyr＋Pro204SerGly157Ser＋Ser160Glu＋Tyr161HisGln 57Asn＋Gly100Glu＋Pro127AsnGly125Pro＋Val197Gly＋Ser210GluVal193Met＋Asn198Ser＋Gly213AsnGly 95Gln＋Pro129Gln＋Thr207GlyGly 59Ser＋Ser126Asp＋Asn212SerGly 63Asn＋Ile159Val＋Ser210GluPhe183Pro＋Tyr208Ile＋Ser210GluLeu 94Ile＋Ile159Glu＋Thr207GlnGln 57Asn＋Gly157Asn＋Leu211CysAla131Gly＋Pro204Asn＋Leu211HisIle159Leu＋Ser206Asp＋Leu211AsnLeu 94Met＋Gly152Ser＋Asn198GluSer182Glu＋Val199Ala＋Leu211ThrLeu124Ser＋Thr207Asp＋Leu211SerVal 93Ser＋Gly125Asn＋Asn198GlnTyr102Pro＋Ala156Ser＋Phe183HisGly 98Gln＋Ser128Glu＋Ile159CysSer104Asp＋Ile105Pro＋Tyr161ThrTyr102Pro＋Ile105Thr＋Ala181GluThr 64Ser＋Ile105Glu＋Gln200AsnSer104Asp＋Gln200Ser＋Thr214ProThr 64Pro＋Ala156Glu＋Thr202GlyGly 63Gln＋Gly100Asp＋Leu211HisAsn 60Ser＋Gly 98Asp＋Leu211ThrTyr161Gly＋Val197Cys＋Ser206GluThr207Gln＋Tyr208Gln＋Ser210GluGly 98Ser＋Ile105Leu＋Ser210GluThr 64Gly＋Pro195Ser＋Thr214SerVal 93Met＋Ser154Asp＋Leu211MetLeu124Ser＋Asn198Ser＋Leu211AspThr 64Gly＋Ala156Ser＋Gly157ProGly 59Ser＋Gln200Asn＋Gly213GluGly 61Asn＋Leu124Ile＋Ala181ProAla 96His＋Gln185Asp＋Leu211AlaSer101Asp＋Asn198Ser＋Thr214SerLeu 94Met＋Asn153Asp＋Pro195GlnPro127Gln＋Gly152Asp＋Val193HisSer 97Asp＋Ile105Thr＋Thr207GlnGly 61Pro＋Ser160Glu＋Thr207GlnLeu124Val＋Pro204Gly＋Leu211IleSer 99Asp＋Asn198Gln＋Gln200SerSer101Glu＋Ala181Asn＋Ala209ProGly 61Asn＋Ala 96Thr＋Gly 98SerVal 93Pro＋Asnl98Asp＋Thr207GlnThr 64Pro＋Ala 96Pro＋Gly155GluThr 64Gly＋Ala181Thr＋Thr214ProThr 64Pro＋Leu 94Cys＋Tyr208MetGln 57Asn＋Leu 94Gln＋Thr214AsnLeu 94Gln＋Tyr161Pro＋Thr207GlyVal 93His＋Gln185Asn＋Thr207ProGly125Pro＋Asn198Glu＋Gln200SerAsn153Ser＋Ile159Cys＋Ser160AspThr 64Gly＋Gln200Glu＋Leu211CysLeu 94Val＋Pro195Gln＋Leu211AspGly 59Pro＋Gly100Ser＋Thr207ProTyr102Cys＋Gly157Gln＋Thr207ProSer 99Asp＋Leu124Ser＋Thr214AsnGly 59Gln＋Gln200Asp＋Thr207SerAla 96His＋Ala181Asp＋Leu211CysVal 93Met＋Ala181His＋Thr207SerPro129Gln＋Asn198Ser＋Thr207SerAsp 58Glu＋Gly 95Ser＋Ala194GlySer160Glu＋Thr207Gln＋Thr214GlyThr 64Asn＋Phe183Asp＋Thr207AsnSer158Asp＋Phe183Ile＋Val197HisGly 95Pro＋Asn153Ser＋Gln200GluGly157Asp＋Pro204Gly＋Thr207SerVal197Pro＋Asn198Gln＋Thr207GlnGly 61Pro＋Leu 94Glu＋Tyr161IlePro129Gln＋Gly155Pro＋Ser210GluVal 93Met＋Ser182Asp＋Gln200SerPro127Ser＋Leu211Met＋Gly213SerAla 96His＋Ser160Asp＋Tyr208GlyGly 98Pro＋Pro129Ser＋Pro204AspGly 59Ser＋Gly 63Pro＋Pro129AspGly100Pro＋Leu124Pro＋Thr207AsnSer103Asp＋Ile159Ala＋Thr202GlnGly 59Asn＋Ser130Glu＋Leu211ThrAla 96Gly＋Ala131Asn＋Gly155ProAsn153Ser＋Gln200Asn＋Ser210GluAsn153Glu＋Thr207Pro＋Leu211AlaPro127Asn＋Val197His＋Thr207AspGly100Asp＋Ala131Gln＋Tyr208ThrGly125Pro＋Phe183Pro＋Asn198GluAsn 60Ser＋Ile105Thr＋Thr207GluThr 64Ser＋Pro127Asp＋Ala131ThrSer 99Asp＋Leu124Asn＋Phe183ProThr 64Gln＋Ser101Asp＋Leu211SerPro195Gln＋Val197Gly＋Leu211SerGly100Asn＋Tyr102Gly＋Thr207ProAsn198Ser＋Gln200Asp＋Leu211ProTyr102Met＋Ser182Glu＋Leu211AlaGly 61Asn＋Gln200Asn＋Gly213AsnSer 99Asp＋Gly152Pro＋Gly157SerLeu124Ser＋Tyr161Met＋Gln200GluTyr102Met＋Leu211Glu＋Thr214SerAsp 58Glu＋Gly 98Asn＋Tyr161AsnIle159Ser＋Gln200Asn＋Tyr208AsnThr 64Asn＋Ala156Gly＋Leu211AspGln200Asn＋Ser210Glu＋Leu211ThrAsn 60Asp＋Gly155Pro＋Val197AsnAla156Gln＋Asn198Glu＋Gly205GlnGln 57Asp＋Gln200Ser＋Tyr208ThrLeu124Ile＋Ala209Glu＋Ser210AspGly 59Glu＋Asn 60Asp＋Leu211AlaAla131Gly＋Gly152Asp＋Asn153GluTyr203Val＋Ser210Glu＋Leu211GluGly152Asn＋Ser210Asp＋Leu211GluPhe183Val＋Ser210Asp＋Leu211AspTyr102Leu＋Ser210Asp＋Leu211GluGln200Asn＋Thr207Glu＋Tyr208GluAla156Ser＋Thr207Glu＋Tyr208AspPro129Asp＋Ser130Asp＋Asn198SerAsn198Ser＋Gln200Glu＋Ser210AspLeu124Cys＋Gln200Glu＋Ser210GluThr 64Gly＋Gln200Asp＋Ser210AspGly 61Asn＋Gln200Asp＋Ser210GluPhe183His＋Val197Glu＋Asn212AspAla 96Gly＋Ser101Glu＋Ser103GluSer126Glu＋Ser128Glu＋Asn198SerGly 61Pro＋Ser154Asp＋Ala156GluVal 93Asp＋Ser104Asp＋Leu211AlaAla 96Ser＋Gly125Asp＋Tyr161AspSer 97Asp＋Ser 99Glu＋Leu211CysSer 97Asp＋Ser 99Glu＋Asn153GlnAla 96Pro＋Gly152Asp＋Ser154AspGly 98Asn＋Asn198Asp＋Gln200AspAsn 60Ser＋Asn198Glu＋Gln200AspPro129Ser＋Asn198Asp＋Gln200AspGln 57Glu＋Gly 59Glu＋Leu211GlnSer128Asp＋Ser130Glu＋Ala209HisSer158Asp＋Ser160Glu＋Gln185AspPro127Gln＋Ser210Glu＋Asn212AspSer210Glu＋Leu211Asn＋Asn212AspVal197Asp＋Gln200Glu＋Ser210AspPhe183Glu＋Tyr208Val＋Gly213AspPro127Glu＋Gly152Glu＋Tyrl61AspLeu 94Glu＋Ser101Asp＋Leu124IleLeu 94Glu＋Ser101Asp＋Gln200AsnPro127Gln＋Asn153Glu＋Ala156GluAsp 58Glu＋Gly 61Gln＋Gly 95AspPro129Gly＋Val197Asp＋Ser210GluAsp 58Glu＋Gly 61Glu＋Gly 95GluVal197Glu＋Ser210Asp＋Gly213AspSer154Glu＋Ser182Glu＋Pro204SerSer101Glu＋Gly125Glu＋Ser126GluSer 99Asp＋Ser104Asp＋Ile105LeuSer128Glu＋Ala131Glu＋Gly196GlnGly 61Glu＋Ser 97Asp＋Ala131HisSer 97Asp＋Gly100Asp＋Ser104GluAsp 58Glu＋Pro204Asn＋Gly205AspGln185Glu＋Leu211Cys＋Gly213AspGln200Asp＋Thr207Glu＋Leu211AsnGly125Ser＋Gln200Asp＋Thr207GluGln200Glu＋Thr207Asp＋Leu211CysAsp 58Glu＋Val 93Gly＋Ser 97GluAsp 58Glu＋Ser 97Glu＋Tyr203AsnGly 61Glu＋Gly 95Asn＋Thr207AspAla 96Pro＋Thr207Asp＋Ser210AspPro204Asn＋Thr207Glu＋Ser210AspGly100Pro＋Thr207Asp＋Ser210AspGly 61Asp＋Thr207Asp＋Ser210AspAsp 58Glu＋Thr 64Asp＋Ser 97GluAsn153Asp＋Gly157Glu＋Ala181GluSer130Asp＋Ser160Asp＋Gly205GlnTyr102Asp＋Ser128Asp＋Ser160AspGln 57Asp＋Ser206Asp＋Thr207GluSer 99Asp＋Ser126Glu＋Gly152GlnAla181His＋Asn198Glu＋Thr214GluGln 57Asn＋Thr 64Glu＋Gln200GluThr 64Asp＋Leu124Thr＋Gln200AspGly 98Asp＋Gly125Glu＋Thr207GlnAsn198Glu＋Gln200Glu＋Pro204GluAsp 58Glu＋Gly 59Asp＋Gln200AspLeu124Asp＋Val197Asp＋Asn198AspPro129Glu＋Asn212Asp＋Gly213GluSer158Asp＋Ser210Asp＋Leu211GluSer101Glu＋Tyr102Glu＋Gln200AspSer103Asp＋Ser104Asp＋Asn198AspAsn 60Glu＋Ser103Glu＋Ser104GluSer182Asp＋Gln200Asp＋Ser210AspGly157Glu＋Gln200Glu＋Ser210AspGln 57Asp＋Gln200Asp＋Ser210GluSer 99Glu＋Gln200Asp＋Ser210AspSer182Glu＋Asn198Asp＋Gln200GluGln 57Ser＋Gln200Asp＋Gly213GluSer 99Glu＋Ser103Asp＋Ile159MetSer 99Asp＋Ser103Asp＋Gly213GlnGly155Glu＋Gln185Glu＋Asn198GluGly155Asp＋Gln185Glu＋Ser210AspGln 57Asp＋Thr 64Glu＋Ser210GluThr207Glu＋Leu211Glu＋Gly213AspSer126Asp＋Ser160Asp＋Ser210AspSer130Glu＋Asn198Asp＋Ser210AspSer126Asp＋Asn198Asp＋Ser210GluSer101Asp＋Asn198Glu＋Leu211GluGly155Glu＋Asn198Asp＋Leu211GluAsp 58Glu＋Asn 60Glu＋Ser103AspSer 99Glu＋Ser104Glu＋Ser126GluGly 59Asp＋Gly 61Gln＋Ser 99AspAsn 60Glu＋Ser158Glu＋Gln185GluGly100Glu＋Ser104Asp＋Ser210GluGly100Asp＋Ser104Asp＋Ser182GluSer128Glu＋Ser130Asp＋Asn198GluSer154Glu＋Gln200Asp＋Leu211GluSer103Asp＋Gln200Glu＋Leu211AspSer103Glu＋Gln200Glu＋Leu211AspSer126Asp＋Ala156Glu＋Ser182AspSer158Asp＋Ser160Glu＋Val197GluSer158Asp＋Ser160Asp＋Val197GluVal 93Gln＋Gly125Glu＋Ser158GluGln 57Asp＋Leu124Asp＋Ser126AspAsn 60Asp＋Ser210Glu＋Asn212GluGly157Asp＋Ser210Glu＋Asn212AspSer 99Glu＋Ser101Glu＋Ser210GluAsp 58Glu＋Asn198Glu＋Pro204GluTyr102Asp＋Ser130Glu＋Ser182AspAsn 60Asp＋Gly 95Glu＋Ser128AspLeu 94Glu＋Ser 97Asp＋Ser206GluGln 57Asp＋Ser126Glu＋Gly152AspLeu 94Glu＋Ser101Glu＋Leu211AspGly 59Glu＋Ser 97Glu＋Ser182GluGly100Asp＋Ser103Glu＋Ser206AspAsn 60Glu＋Gln185Glu＋Thr214GluAsn 60Glu＋Ser130Glu＋Pro204AspAsn 60Asp＋Asn198Asp＋Pro204AspSer 99Asp＋Ser182Glu＋Gln185AspPro204Asp＋Ser210Glu＋Leu211GlyTyr161Val＋Pro204Glu＋Ser210GluSer101Glu＋Asn198Asp＋Gly213GluLeu124Asp＋Ser154Glu＋Val193SerSer 99Glu＋Ala181Glu＋Asn212AspAla 96Asp＋Gly100Asp＋Ser154GluGly 61Asp＋Ser 99Asp＋Ser210GluAsp 58Glu＋Thr207Glu＋Ser210Asp表35多个环状区的四突变的突变体Gln 57Ser＋Asn 60Ser＋Leu 94Gly＋Gln200GluVal 93Gln＋Tyr102Cys＋Thr207Gly＋Gly213AspIle105Val＋Leu124Ile＋Ser154Glu＋Asn198GlnLeu 94Gly＋Ser103Glu＋Gln200Asn＋Ala209GlyAla156His＋Gln200Ser＋Thr207Pro＋Leu211ThrPro129Asn＋Gly155Asp＋Ala209Gln＋Asn212GlnLeu124Pro＋Ala194Asn＋Asn198Glu＋Gln200AsnGly 61Ser＋Asn198Gln＋Gln200Ser＋Leu211HisTyr203Thr＋Thr207Asn＋Tyr208Gly＋Asn212GluGln 57Asn＋Asn153Asp＋Leu211Ser＋Gly213ProAla131Asn＋Gln185Ser＋Val193Ser＋Gln200AsnAla181Gly＋Ala194Pro＋Asn198Ser＋Asn212AspSer101Asp＋Leu124Val＋Pro195Gln＋Thr207ProGly 95Asp＋Asn198Gln＋Gln200Ser＋Thr207SerAla 96Gln＋Tyr102Pro＋Ser182Glu＋Val197AsnAsn 60Gln＋Gly100Ser＋Ser101Asp＋Ile159ProGly 63Asn＋Asn153Asp＋Gly205Ser＋Thr207SerSer126Glu＋Ala131Pro＋Asn153Ser＋Phe183ThrAsn153Gln＋Gly205Pro＋Ser210Glu＋Gly213ProIle159Glu＋Ala194Ser＋Pro204Gly＋Leu211IleAsp 58Glu＋Val 93Ser＋Ala194Pro＋Gln200SerGly 95Asp＋Ile105Gln＋Phe183Cys＋Asn198GlnGly 59Pro＋Gly 98Asn＋Leu211Val＋Thr214AspGly 61Pro＋Val 93Pro＋Pro129Asn＋Gly213ProGln 57Asn＋Ala156Pro＋Ala181Pro＋Phe183GlyGly 95Gln＋Gly152Asp＋Phe183Met＋Asn198GlnGly 59Ser＋Gly 95Asn＋Pro127Gln＋Ser210GluVal197Cys＋Asn198Gln＋Thr207Glu＋Leu211CysPro129Asn＋Ser154Asp＋Phe183Pro＋Leu211CysGly 59Gln＋Gly100Glu＋Ala131Gly＋Ala181GlnGly 61Glu＋Tyr161Leu＋Pro204Ser＋Gly213ProAsp 58Glu＋Ile159Met＋Tyr161Gln＋Gly213AsnIle105Pro＋Leu124Asn＋Ile159Ser＋Asn198AspTyr102Pro＋Ala156Ser＋Phe183His＋Pro204AspTyr102Pro＋Ile105Pro＋Gln200Asn＋Pro204GluThr 64Ser＋Ile105Glu＋Gln200Ser＋Thr214ProGly 63Gln＋Gly100Asp＋Asn198Ser＋Leu211HisAsn 60Asp＋Ile105Leu＋Gly152Gln＋Ala209GlnGly152Gln＋Gln185Asn＋Gly205Ser＋Leu211AlaGln 57Ser＋Gly100Pro＋Pro127Gly＋Asn198GluLeu 94Cys＋Pro127Gly＋Val197Gly＋Asn198SerLeu 94Thr＋Tyr102Cys＋Leu124Cys＋Tyr208SerAla 96Ser＋Ala181Asn＋Pro204Ser＋Tyr208IleLeu124Val＋Asn198Gln＋Pro204Gly＋Leu211IleGly 61Asn＋Ala 96Thr＋Gly 98Ser＋Ser210AspThr 64Pro＋Ala 96Pro＋Gly155Glu＋Leu211CysGly157Asp＋Val197Pro＋Asn198Gln＋Pro204GlyPro129Gln＋Gly155Pro＋Val197His＋Ser210GluVal 93Met＋Ser182Asp＋Gln200Ser＋Gly213SerLeu 94Glu＋Pro127Ser＋Gln200Asn＋Leu211MetAla 96His＋Gly 98Pro＋Pro129Ser＋Pro204AspSer 97Asp＋Gly100Pro＋Leu124Pro＋Thr207AsnGly 59Asn＋Thr207Pro＋Ser210Glu＋Leu211ThrAla 96Gly＋Ala131Asn＋Gly155Pro＋Ser210GluAsn153Ser＋Val197Thr＋Gln200Asn＋Ser210GluAsn 60Ser＋Ile105Thr＋Ser182Glu＋Thr207SerGly 98Asn＋Tyr161Asn＋Gln200Asn＋Tyr208AsnLeu 94Gly＋Ser 99Glu＋Leu124Ser＋Ala181ProSer104Glu＋Ala131Pro＋Leu211Ser＋Gly213SerTyr102Asp＋Asn198Ser＋Thr207Gln＋Leu211AsnGly 98Pro＋Ser206Glu＋Leu211Gly＋Gly213ProAla131Asn＋Ala181Thr＋Tyr208Gln＋Ser210AspGly155Gln＋Tyr161Cys＋Gln185Ser＋Ala209GlnThr 64Asn＋Val 93Asn＋Ser126Glu＋Gln200AsnAla131Pro＋Phe183Ala＋Thr207Asp＋Thr214GlnSer 97Glu＋Leu124Pro＋Ala131Ser＋Pro204GlyGly 59Asp＋Tyr161His＋Thr207Gly＋Leu211MetGly 95Pro＋Gly 98Pro＋Ser101Glu＋Leu211CysGly 59Ser＋Thr 64Gly＋Leu124Ile＋Gly125GluThr 64Pro＋Ser101Glu＋Tyr161Gly＋Gln200AsnGly100Glu＋Pro195Ser＋Val197Ser＋Thr207AsnSer160Glu＋Tyr161Gln＋Pro204Ser＋Leu211GlnLeu124Met＋Tyr161Leu＋Ser182Glu＋Leu211HisIle105Thr＋Pro127Asn＋Gly157Asn＋Tyr161ProGln185Asn＋Thr207Gln＋Asn212Ser＋Thr214GlnVal 93Gln＋Leu 94Ser＋Gly 98Glu＋Thr214GlyAsn153Gln＋Ser154Glu＋Ala181Asn＋Leu211ValGly125Asn＋Ser154Asp＋Val197Ser＋Leu211ValIle159Asn＋Gln185G1u＋Val197Met＋Asn212GlnAla 96Pro＋Gly196Gln＋Val197Ala＋Asn198GlnAla131Glu＋Gly152Gln＋Ala194Ser＋Leu211IlePhe183Ser＋Asn198Asp＋Gln200Ser＋Thr207ProGly 95Gln＋Gly100Ser＋Ser130Glu＋Thr207GlnLeu124Cys＋Gly152Ser＋Tyr161Met＋Thr207GlnLeu 94Met＋Leu124Met＋Leu211Ala＋Asn212SerGly 61Asn＋Ala 96Ser＋Gly125Pro＋Gln200AspIle159Gln＋Val197Glu＋Gln200Asn＋Tyr208HisTyr161Asn＋Ser182Glu＋Ala209Ser＋Asn212SerGln 57Asn＋Gly152Ser＋Phe183Glu＋Gln185AsnAsn198Gln＋Thr207Glu＋Ala209Gly＋Gly213ProVal197Glu＋Tyr208Gly＋Ala209Pro＋Leu211ProAla131His＋Gly205Gln＋Tyr208Asp＋Leu211GlnGly 61Asn＋Val 93Cys＋Gly 98Gln＋Leu211GluLeu124Ala＋Val197Gln＋Ser210Glu＋Asn212SerPro127Gln＋Ile159Leu＋Ser160Glu＋Ala209SerAsp 58Glu＋Gly 59Gln＋Gly205Gln＋Leu211CysAsp 58Glu＋Phe183Tyr＋Asn198Ser＋Thr207GlnPro204Gly＋Ser206Asp＋Thr207Gln＋Leu211HisGln 57Asp＋Gly 98Ser＋Ile105Gln＋Tyr208MetSer103Glu＋Gly155Ser＋Ala209His＋Leu211ThrSerl04Glu＋Ala181Asn＋Asn198Gln＋Leu211ThrGln185Asn＋Val197Cys＋Gly205Asp＋Leu211CysGly 59Gln＋Tyr102Glu＋Pro127Gly＋Ala209ThrAla131Thr＋Asn153Ser＋Val193Gln＋Val197AspAla131Ser＋Asn153Ser＋Phe183Tyr＋Ala194SerPro129Gln＋Ala181Pro＋Ala209His＋Leu211AlaPro127Gly＋Gly196Asn＋Ser210Glu＋Leu211MetLeu 94Ile＋Gly100Asp＋Asn198Ser＋Tyr208LeuAsn153Ser＋Gly155Pro＋Gly205Asp＋Leu211SerGly 63Asn＋Gly152Pro＋Asn198Gln＋Thr207GluGly152Ser＋Ser160Glu＋Tyr161His＋Leu211MetPro129Asn＋Asn153Gln＋Gly155Gln＋Ser210AspGln 57Ser＋Thr 64Gln＋Val197Asn＋Gln200AspSer126Asp＋Asn153Gln＋Pro204Gln＋Leu211GlnGly 61Gln＋Ala 96Gln＋Phe183Leu＋Leu211ThrGly 63Asn＋Ala156Asn＋Asn198Ser＋Ser210GluGly 59Asp＋Gly205Pro＋Thr207Gln＋Leu211AsnGly 98Ser＋Leu124Pro＋Ile159Thr＋Val197GluGly205Pro＋Ala209Gln＋Ser210Glu＋Leu211ProPro129Ser＋Thr207Gly＋Ser210Glu＋Leu211AsnGln 57Asp＋Ala131Gln＋Leu211Met＋Asn212GlnThr 64Asn＋Tyr161Ile＋Phe183Asn＋Ser210AspGly155Gln＋Pro204Ser＋Ala209His＋Leu211AspGly100Gln＋Gly157Pro＋Ile159Leu＋Ser160AspGly 98Pro＋Ser103Asp＋Ala131Gly＋Leu211AsnGln 57Ser＋Gly 61Pro＋Ile159Gln＋Leu211AsnGly155Ser＋Gln185Asp＋Val199Met＋Thr207GlyAsp 58Glu＋Ala 96Gly＋Phe183Thr＋Thr207GlyVal 93Ala＋Ser126Glu＋Ala131Thr＋Thr207ProThr 64Gln＋Gly 98Pro＋Val197Thr＋Gln200GluGly152Glu＋Asn153Gln＋Gly155Pro＋Gly213ProSer154Glu＋Asn198Ser＋Gln200Ser＋Leu211SerIle105Thr＋Ile159Thr＋Phe183Val＋Leu211AspLeu124Pro＋Ser128Glu＋Ile159Met＋Leu211ProGln 57Asp＋Asp 58Glu＋Leu211Thr＋Thr214SerGly 59Asp＋Asn 60Asp＋Val197Pro＋Val199AlaAla 96Thr＋Leu124Cys＋Val197Asp＋Asn198GluThr 64Asn＋Ala156Gly＋Ser210Asp＋Leu211AspLeu124Ser＋Gln200Asn＋Gly205Asp＋Ser206GluGly 63Pro＋Ala156Asp＋Gly157Asp＋Ile159GlnThr 64Gly＋Gly100Glu＋Ser101Glu＋Leu211AsnGly 61Asn＋Asn153Gln＋Gln200Asp＋Ser210AspLeu124Ser＋Gln200Glu＋Ser210Glu＋Leu211GlnVal197Gln＋Gln200Glu＋Thr207Ser＋Ser210GluAla209Glu＋Ser210Glu＋Leu211G1u＋Gly213SerThr207Pro＋Ser210Glu＋Leu211Asp＋Asn212GluSer101Asp＋Ser104Asp＋Asn198Gln＋Leu211SerThr 64Asn＋Ala 96Asp＋Gly 98Asp＋Gly213AsnAsn198Glu＋Gln200Asn＋Ser210Asp＋Leu211ThrTyr161Val＋Asn198Glu＋Ser210Glu＋Gly213AsnVal 93Pro＋Asn198Glu＋Gln200Ser＋Ser210GluVal193Pro＋Asn198Asp＋Thr207Ser＋Ser210AspGln185Ser＋Asn198Asp＋Gln200Asp＋Ser210GluGly 95Ser＋Gly 98Gln＋Asn198Asp＋Leu211AspVal 93His＋Ala 96His＋Gln200Glu＋Ala209AspLeu 94Cys＋Ile159Leu＋Gln200Asp＋Ala209GluGln 57Asn＋Gly 61Asp＋Thr 64Asp＋Leu 94ValAla156Gln＋Asn198Glu＋Gly205Gln＋Asn212AspTyr102Asp＋Ser104Asp＋Phe183Gln＋Val197HisAla 96Ser＋Asn153Ser＋Gly155Asp＋Ser182AspAla 96Asn＋Asn153Glu＋Gln185Glu＋Thr214GlyAsp 58Glu＋Gly 61Glu＋Asn153Ser＋Leu211CysPro129Ser＋Asn198Asp＋Gln200Asp＋Leu211ProGly125Ser＋Asn198Asp＋Gln200Glu＋Leu211AlaAsn198Asp＋Gln200Asp＋Thr207Pro＋Leu211ProGly 98Pro＋Asn198Asp＋Gln200Glu＋Tyr203SerAsn198Glu＋Gln200Glu＋Tyr203Leu＋Thr214AsnGly100Asp＋Tyr102Asp＋Asn153Ser＋Tyr208LeuThr 64Gln＋Thr207Asn＋Ser210Glu＋Asn212GluPro127Gln＋Ile159Cys＋Ser210Glu＋Asn212AspVal 93Ala＋Ser 99Asp＋Ser101Glu＋Ile105AsnGly100Gln＋Gly155Gln＋Asn212Asp＋Thr214AspGly 95Ser＋Tyr102Leu＋Pro129Glu＋Ser160AspGly 59Ser＋Pro129Asp＋Ser160Asp＋Leu211ProThr 64Pro＋Ala 96Thr＋Asn153Glu＋Ser182AspAsp 58Glu＋Gly 61Gln＋Gly 95Asp＋Ala181ThrGly 63Pro＋Ser 99Asp＋Ser101Glu＋Ser103GluAsn153Gln＋Val197Asp＋Leu211Glu＋Thr214GluGly152Ser＋Ala181Pro＋Asn198Asp＋Gly213AspSer154Glu＋Gly155Pro＋Ser160Asp＋Asn198GlnGly152Asn＋Ser154Glu＋Ser158Glu＋Leu211AsnAsn 60Glu＋Leu 94Asp＋Tyr208Ala＋Gly213ProThr 64Asp＋Gln200Asp＋Ser210Asp＋Gly213ProSer103Asp＋Ser130Glu＋Ile159Ala＋Thr202GlnSer103Glu＋Ser130Glu＋Asn198Gln＋Pro204AsnThr207Pro＋Ser210Asp＋Leu211Glu＋Thr214GluSer210Glu＋Leu211Met＋Gly213Glu＋Thr214GluGly125Ser＋Gln200Asp＋Ser206Glu＋Thr207GluAla 96Pro＋Gly125Asp＋Ser154Asp＋Ala156AspSer 99Asp＋Ser104Asp＋Ile105Mer＋Gln200SerGly 59Gln＋Ser128Asp＋Ser154Glu＋Ser160AspAsp 58Glu＋Gly 98Asp＋Leu124Ser＋Tyr161MetGln185Glu＋Asn198Ser＋Leu211Cys＋Gly213AspGly 61Gln＋Gly 98Pro＋Gln200Asp＋Thr207GluGly100Gln＋Gln200Glu＋Tyr203Pro＋Thr207GluVal197Pro＋Gln200Asp＋Thr207Asp＋Ala209GlnAsp 58Glu＋Val 93Gly＋Ser 97Glu＋Tyr161IleAsp 58Glu＋Ser 97Asp＋Gln200Ser＋Tyr208ThrGly152Asp＋Gly157Glu＋Val199Ser＋Thr207SerVal197Gln＋Pro204Ser＋Thr207Asp＋Ser210AspGly125Asn＋Pro129Asn＋Thr207Glu＋Ser210GluSer101Asp＋Ser128Asp＋Pro129Glu＋Gly152SerLeu 94Ala＋Asn198Glu＋Gln200Asp＋Thr214GluAla 96Ser＋Ile105Glu＋Ser126Glu＋Ser160AspSer130Asp＋Ser158Glu＋Ser160Asp＋Asn198GlnLeu124AsP＋Phe183Glu＋Tyr208Val＋Gly213AspSer101Asp＋Ser126Glu＋Ala156Gly＋Thr207AsnAsp 58Glu＋Asn 60Asp＋Gly100Asp＋Ala181GlnAsp 58Glu＋Gly205Ser＋Thr207Asp＋Tyr208GluThr 64Gln＋Leu 94Glu＋Ser 97Asp＋Gly125AspSer158Glu＋Ile159Asp＋Leu211Val＋Gly213GluGln 57Asp＋Thr 64Asp＋Leu124Thr＋Gln200AspGly 61Glu＋Gln200Ser＋Gly205Glu＋Ser210GluAsp 58Glu＋Ile105Gly＋Leu211Asp＋Asn212GluGln 57Asp＋Asp 58Glu＋Gln200Glu＋Pro204SerGln 57Asp＋Asp 58Glu＋Ala 96His＋Ser160GluGly157Asp＋Gln200Ser＋Ala209Glu＋Ser210GluGly 59Glu＋Asn 60Asp＋Gly 95Asn＋Gln185GluSer101Asp＋Tyr102His＋Val197Glu＋Asn198GluGly152Asp＋Asn153Glu＋Ser210Asp＋Leu211MetGly 59Asp＋Tyr102Ile＋Ser210Asp＋Leu211AspSer101Glu＋Tyr102Asp＋Gln200Glu＋Leu211AsnGly 59Asn＋Ser130Glu＋Gly205Asp＋Ser206AspSer128Asp＋Tyr161Gly＋Ser206Asp＋Thr207GluAsn 60Gln＋Ser 97Glu＋Ser182Asp＋Phe183AspGly 98Glu＋Ser 99Asp＋Ser160Glu＋Thr207GlnGly 98Asp＋Thr202Gln＋Thr207Glu＋Tyr208AspSer154Glu＋Tyr161Leu＋Ser210Glu＋Asn212GluAsn 60Asp＋Ile105Ser＋Gln200Asp＋Ser210GluAla131Asp＋Gln200Glu＋Ser210Asp＋Leu211ProGln 57Asp＋Gln200Glu＋Ser210Asp＋Gly213ProPro129Gln＋Ser160Glu＋Gln200Asp＋Ser210GluPro129Asp＋Asn198Gln＋Gln200Asp＋Ser210GluGly 59Glu＋Thr 64Gly＋Gln200Glu＋Ser210GluAsn 60Glu＋Ala 96Glu＋Asn198Gln＋Ser210GluAsn 60Asp＋Ala 96Asp＋Ser130Glu＋Ala156ProGln 57Ser＋Ile159Ser＋Gln200Asp＋Gly213GluGly155Asp＋Gln185Glu＋Asn198Glu＋Tyr203HisSer104Glu＋Leu124Asp＋Gln185Ser＋Val197GlyGly 59Gln＋Ser101Glu＋Ser103Asp＋Asn198AspAsn 60Gln＋Ser126Asp＋Ser128Asp＋Ser210GluSer126Glu＋Ser128Asp＋Gly155Asn＋Asn212GluSer103Asp＋Asn198Glu＋Ser210Glu＋Thr214GlnSer160Glu＋Asn198Glu＋Ser210Asp＋Leu211SerAsp 58Glu＋Asn198Asp＋Thr207Pro＋Ser210AspGly 98Asn＋Ser103Glu＋Asn198Glu＋Ser210GluSer128Asp＋Gly155Pro＋Asn198Asp＋Ser210AspGly152Glu＋Asn198Asp＋Thr207Asn＋Ser210AspSer101Asp＋Pro129Gly＋Asn198Glu＋Leu211GluAsn198Glu＋Tyr208Asp＋Gly213Ser＋Thr214SerSer 97Glu＋Gly152Glu＋Ser154Asp＋Gln185SerThr 64Asp＋Ser210Asp＋Leu211Val＋Gly213GlnThr 64Glu＋Gly205Ser＋Ser210Asp＋Leu211AsnLeu 94Thr＋Ser126Glu＋Pro204Glu＋Thr207AspGly100Gln＋Tyr102Asp＋Pro204Asp＋Thr207AspSer101Glu＋Asn198Glu＋Gln200Glu＋Leu211AlaThr 64Asp＋Ser 99Glu＋Pro204Glu＋Thr207AsnGly155Pro＋Ser182Asp＋Gln200Asp＋Leu211GluGly100Asn＋Ser160Glu＋Gln200Glu＋Leu211GluAsp 58Glu＋Val 93Asn＋Ser206Glu＋Leu211MetAla131Pro＋Asn153Glu＋Ser182Asp＋Asn198GluAsp 58Glu＋Ala 96Glu＋Pro127Ser＋Gly157GluLeu 94Asp＋Tyr102Leu＋Thr207Glu＋Ala209GluAla 96Gln＋Ser158Asp＋Ser160Glu＋Val197GluGly100Asp＋Ser126Glu＋Ser154Asp＋Ile159GlyTyr102Asn＋Gln200Glu＋Thr207Glu＋Gly213AspVal 93Glu＋Pro127Ser＋Ser210Asp＋Asn212AspIle105Asp＋Ala131Glu＋Ala156Gln＋Thr202ProGln 57Ser＋Ser126Glu＋Gly152Asp＋Ser210GluSer126Glu＋Gly152Glu＋Asn198Glu＋Thr207AsnLeu 94Glu＋Ser101Asp＋Tyr102Leu＋Thr207GluLeu 94Asp＋Ser101Asp＋Ser154Asp＋Gly213AsnAsn 60Asp＋Thr 64Glu＋Ser103Glu＋Thr214AsnAsp 58Glu＋Val197Asp＋Ser210Glu＋Leu211ProSer160Asp＋Phe183Tyr＋Val197Asp＋Ser210AspPro129Glu＋Val197Glu＋Ser210Glu＋Leu211HisSer126Asp＋Ser130Glu＋Ser182Glu＋Gln200SerSer126Asp＋Ser130Glu＋Asn198Asp＋Leu211H1sThr 64Ser＋Pro204Asp＋Ser210Asp＋Leu211IleSer 97Asp＋Gly100Glu＋Gly152Pro＋Asn198GluSer 97Asp＋Gly100Glu＋Gln200Glu＋Tyr208GlyAsp 58Glu＋Gly 59Pro＋Ser154Asp＋Gly157AspGly155Gln＋Ala181Glu＋Gln185Asp＋Ser210AspGly125Glu＋Asn153Asp＋Gln200Asp＋Tyr208ProGln 57Ser＋Ser101Asp＋Ser154Asp＋Ser160AspSer104Asp＋Asn198Glu＋Ala209Asp＋Leu211MetLeu124Ile＋Gly125Pro＋Pro129Glu＋Ser158GluSer154Glu＋Ser158Asp＋Gly205Pro＋Thr207GluGly152Glu＋Ser158Asp＋Gln200Asp＋Thr207ProAla131Asp＋Gly152Asp＋Ser158Asp＋Val197ThrSer103Glu＋Gln200Glu＋Thr207Pro＋Asn212AspSer130Glu＋Gln200Glu＋Thr202Ser＋Asn212GluAsn 60Glu＋Leu 94Glu＋Ile105Leu＋Ser130GluSer 99Glu＋Pro127Asp＋Pro129Ser＋Ser154GluAsn153Glu＋Ile159Gly＋Leu211Glu＋Gly213AsnAla156Asp＋Ser160Glu＋Gly205Gln＋Leu211AspSer104Asp＋Ser130Asp＋Gly155Pro＋Gly157AsnGly 98Asp＋Ser101Asp＋Ser128Asp＋Ala181GlnAsn 60Asp＋Tyr102Gly＋Gln200Glu＋Thr207AspGly 59Pro＋Ala131Asp＋Ser154Asp＋Thr214AspSer103Glu＋Ser130Glu＋Asn198Glu＋Asn212GlnAsn 60Asp＋Gly 98Asn＋Ser103Glu＋Ser130AspSer160Asp＋Phe183Ser＋Ser210Glu＋Thr214AspAsp 58Glu＋Ser 99Asp＋Val197Pro＋Pro204AsnAsp 58Glu＋Gly 98Glu＋Gly155Gln＋Leu211AspThr 64Glu＋Leu 94Asp＋Gln185Ser＋Ser210AspAsn 60Asp＋Ser 99Glu＋Ala181Glu＋Val197AsnSer128Glu＋Ser206Asp＋Ala209Glu＋Asn212GlnGly100Asp＋Gln185Glu＋Leu211Ser＋Gly213GluSer126Glu＋Ser154Glu＋Asn198Gln＋Gly205GluGly125Glu＋Val197Thr＋Gln200Glu＋Thr207AspASn 60Glu＋Ile105Pro＋Ile159Val＋Leu211GluPro129Asn＋Gln200Glu＋Pro204Asp＋Thr207GlnAsn198Ser＋Gln200Asp＋Tyr203Ile＋Pro204GluGly 98Ser＋Ser103Glu＋Ala131Glu＋Ser210AspSer103Glu＋Ala131Asp＋Tyr161His＋Ser210GluGly 61Glu＋Gly125Pro＋Ser126Glu＋Thr207GluGlV152Asp＋Gly157Asp＋Pro204Asp＋Ala209ThrPro127Asp＋Asn153Glu＋Pro204Ser＋Ser206AspGln 57Asp＋Gln200Ser＋Thr207Glu＋Ser210GluSer154Asp＋Thr207Glu＋Ser210Glu＋Thr214GlnSer130Glu＋Pro204Asn＋Thr207Glu＋Ser210AspAsp 58Glu＋Val 93Ala＋Ser101Glu＋Ser130AspGly125Glu＋Ser130Asp＋Gln200Asp＋Leu211ThrLeu 94Glu＋Ser182Asp＋Thr207Gln＋Asn212GluSer 97Glu＋Ile159Asp＋Phe183Ile＋Thr214AspGly 95Glu＋Ser103Asp＋Pro129Asn＋Ala131ProAsn 60Gln＋Gly 61Glu＋Ser130Asp＋Ser210AspSer101Glu＋Ser126Asp＋Ala209Glu＋Asn212SerSer101Glu＋Ser126Asp＋Tyr161Leu＋Gly205GluAsn 60Ser＋Gly 98Asp＋Ser126Glu＋Leu211ThrVal 93Glu＋Ser 97Glu＋Pro129Ser＋G1n200GluThr 64Asn＋Gln200Asn＋Thr207Asp＋Leu211AspGly 98Glu＋Ser104Asp＋Ser126Asp＋Phe183GlyGly125Asp＋Gly157Asp＋Thr207Gln＋Gly213GluLeu 94Asp＋Ser128Asp＋Gly152Asp＋Leu211MetSer158Glu＋Ser182Glu＋Thr207Gly＋Tyr208CysGln200Asn＋Pro204Gln＋Ser210Glu＋Thr214GluThr 64Glu＋Ala131Asp＋Ser210Asp＋Leu211AlaAsp 58Glu＋Gly 59Pro＋Gly100Pro＋Thr207AspAsn198Gln＋Gln200Glu＋Ser206Glu＋Tyr208HisGly 95Pro＋Ser126Asp＋Ala131Glu＋Gln200GluSer 97Glu＋Ser101Glu＋Gln200Asp＋Pro204AsnSer 97Asp＋Ser101Asp＋Gly125Gln＋Ser210AspAla131Gly＋Ser154Asp＋Tyr161Glu＋Ser210AspSer126Glu＋Pro204Asp＋Thr207Gly＋Ser210AspGly 98Glu＋Ser104Asp＋Val197Glu＋Asn198SerSer104Asp＋Ser130Glu＋Ala156Thr＋Thr207AspSer104Asp＋Ser130Asp＋Ala131Thr＋Thr207AspPro127Ser＋Ser158Asp＋Gln200Ser＋Thr214GluAsn 60Asp＋Tyr102Ser＋Val197Asn＋Thr207AspSer101Asp＋Pro127Ser＋Ala131Glu＋Ala156GluAsp 58Glu＋Ser 99Asp＋Gln200Glu＋Tyr208SerAsp 58Glu＋Ser 99Asp＋Leu124His＋Asn198GluGly 61Glu＋Thr 64Ser＋Val 93Asp＋Tyr208GlyAsn 60Glu＋Gln185Ser＋Asn198Ser＋Ser206AspGly 61Pro＋Ser158Asp＋Alal81Glu＋Leu211AspTyr161Glu＋Gln200Glu＋Tyr203His＋Pro204GluGly 61Glu＋Gly157Asp＋Leu211Met＋Asn212AspGly 59Ser＋Gly 98Glu＋Tyr102Glu＋Ser158Asp
表36多个环状区的五突变的突变体Val 93Gln＋Tyr102Cys＋Ser154Glu＋Asn198Gln＋Thr207GlyLeu 94Gly＋Leu124Ile＋Asn198Gln＋Ala209Gly＋Ser210GluLeul24Asp＋Ala156His＋Gln200Ser＋Thr207Pro＋Leu211ThrPro129Asn＋Gly155Asp＋Gly157Asn＋Thr207Ser＋Asn212GlnGly 61Ser＋Gly 98Glu＋Pro127Asn＋Gln200Ser＋Thr207ProAla156Gln＋Gln185Ser＋Gln200Asn＋Ser210Asp＋Leu211HisGly 63Asn＋Tyr102Pro＋Ala156Gln＋Ser182Glu＋Val197AsnAla 96Pro＋Ile105Pro＋Thr207Ser＋Leu211Ile＋Thr214ProVal 93Asn＋Val197Gly＋Asn198Gln＋Ser210Asp＋Leu211SerVal 93Pro＋Leu 94Ile＋Ile159Asn＋Gln200Ser＋Thr207SerGly 61Pro＋Gly 95Pro＋Ser154Glu＋Ala181His＋Gly213ProGly157Ser＋Ser182Glu＋Pro195Gln＋Thr207Pro＋Leu211SerSer 97Glu＋Tyr102Met＋Asn198Ser＋Ala209Gln＋Leu211AlaLeu124His＋Ala131His＋Gly157Pro＋Asn198Asp＋Thr202SerGly 95Asp＋Ala181Pro＋Gln200Asn＋Leu211Ala＋Gly213GlnLeu 94Ile＋Gly 95Asn＋Ser158Glu＋Leu211Met＋Gly213SerGly 98Asn＋Gly125Pro＋Pro127Gly＋Tyr161Ile＋Tyr208MetAla 96Ser＋Ser154Glu＋Tyr161Gly＋Gly196Gln＋Val197GlnLeu 94Ser＋Gly 95Asn＋Pro127Asn＋Val197Ser＋Thr214AsnAsp 58Glu＋Val 93Gln＋Leu 94Met＋Gln200Asn＋Thr207GlnGly 95Asp＋Gly155Ser＋Val197Thr＋Thr207Gly＋Leu211ProGly 98Ser＋Ala131Glu＋Val197Gln＋Gln200Ser＋Pro204GlyGln 57Asn＋Val 93Thr＋Tyr102Ser＋Leu124Pro＋Ser126AspSer101Asp＋Tyr102Cys＋Leu124Val＋Phe183Gly＋Val199AsnAsp 58Glu＋Thr 64Gln＋Ile105Pro＋Gly125Gln＋Asn198SerGly155Glu＋Tyr161Val＋Pro204Gly＋Thr207Pro＋Leu211IleGly100Gln＋Ser182Asp＋Phe183Asn＋Asn198Ser＋Thr207GlnGly 98Glu＋Pro129Gly＋Gly155Pro＋Thr207Gly＋Thr214GlyLeu 94Met＋Glyl00Ser＋Gly196Gln＋Gln200Asn＋Asn212AspSer104Glu＋Gly152Gln＋Asn153Gln＋Ala194Gln＋Asn212SerPro129Gln＋Val197Thr＋Tyr208Ile＋Ser210Glu＋Thr214SerVal 93Gly＋Ala 96Ash＋Pro129Gln＋Ser160Asp＋Pro204GlyPro127Gln＋Ala131Asn＋Gly196Ser＋Val197Thr＋Ser206GluPro127Ser＋Ser128Asp＋Ala156His＋Ala194Gln＋Leu211AlaGly 95Gln＋Pro127Gly＋Val197Gly＋Gly205Asn＋Ser210GluAla131His＋Pro204Gly＋Thr207Asn＋Leu211Thr＋Gly213AsnLeu 94Gly＋Tyr102Gln＋Phe183Val＋Thr207Ser＋Leu211AspGly 61Asn＋Ile105Met＋Gly125Ser＋Val193Gly＋Asn198GluGly 59Pro＋Ala 96Gly＋Ser101Asp＋Tyr208His＋Leu211HisTyrl02Val＋Gln200Asp＋Pro204Ser＋Leu211Ile＋Thr214ProVal 93Thr＋Ser104Glu＋Pro195Ser＋Val199Pro＋Leu211ValIle159Gln＋Tyr161Ala＋Phe183Leu＋Ser210Glu＋Thr214GlyGly 59Asn＋Pro127Ser＋Ile159Ala＋Gln200Asn＋Leu211MetGln 57Glu＋Val 93His＋Gly125Gln＋Ala131Thr＋Ala156AsnSer126Asp＋Gly157Ser＋Asn198Gln＋Gln200Ser＋Thr207SerGly125Asn＋Pro129Ser＋Phe183Asp＋Thr207Gly＋Leu211GlyAsn 60Asp＋Leu 94Met＋Tyr102Pro＋Gly152Asn＋Thr207GlnIle105Leu＋Gly152Pro＋Ser158Glu＋Asn198Ser＋Thr214GlyAla 96Asn＋Ala131Thr＋Gly157Asn＋Thr207Gly＋Ser210GluVal 93Gln＋Leu124Met＋Alal81Gln＋Asn198Glu＋Gln200SerPro127Ser＋Ile159Thr＋Pro195Gly＋Asn198Ser＋Gln200GluAla 96Asn＋Val197His＋Asn198Ser＋Gln200Asn＋Asn212AspAsp 58Glu＋Leu 94Ala＋Leu124Pro＋Ala156Gly＋Leu211GlnIle105Thr＋Gly157Pro＋Phe183Pro＋Val197Asp＋Tyr208ValGly 95Glu＋Gly100Gln＋Tyr102Gly＋Tyr161His＋Tyr208CysThr 64Gly＋Val 93Asp＋Gly 95Pro＋Gly100Pro＋Asn198GlnGln 57Asn＋Asn 60Gln＋Ser101Glu＋Gly155Ser＋Pro204SerGly125Ser＋Gln185Asn＋Asn198Gln＋Ser206Glu＋Ala209HisGly 98Ser＋Pro129Asn＋Val197Glu＋Gln200Asn＋Tyr208AsnGly 98Asp＋Asn153Gln＋Gln200Asn＋Pro204Ser＋Leu211SerAla 96Thr＋Ala181Thr＋Val197Asn＋Gln200Asn＋Thr207ProGly 95Gln＋Pro129Asn＋Tyr161Ala＋Gln185Asp＋Thr207AsnGly 95Asn＋Gly152Gln＋Ser160Glu＋Ala194Asn＋Tyr208HisGln 57Asp＋Phe183Leu＋Thr207Ser＋Ala209Gln＋Leu211MetIle105His＋Gly125Asp＋Gly152Asn＋Pro204Asn＋Tyr208LeuGln 57Asn＋Leu 94Glu＋Ala 96Gln＋Pro127Gly＋Leu211ProAsn153Ser＋Tyr161Met＋Ala181Gln＋Asn198Ser＋Ser210AspGly 95Asn＋Gly 98Glu＋Pro129Asn＋Ala209Pro＋Thr214ProGly 61Asn＋Asn198Ser＋Ala209Glu＋Leu211Ser＋Gly213ProSer 99Asp＋Pro127Asn＋Pro195Asn＋Tyr208His＋Leu211CysAsn 60Gln＋Thr 64Asn＋Gly100Asn＋Ile105Leu＋Pro127AsnPro127Glu＋Ala194Gln＋Gln200Asn＋Thr207Ser＋Leu211CysThr 64Gly＋Leu 94Gly＋Val197Asn＋Gln2005er＋Ala209ProGly 59Gln＋Leu 94Ser＋Ala 96Ser＋Leu211Thr＋Thr214GluPro129Gly＋Ala194Asn＋Gln200Ser＋Leu211Gln＋Asn212AspAla131Gly＋Asn153Glu＋Thr207Asn＋Leu211Thr＋Gly213GlnGly 59Pro＋Thr 64Asn＋Asn198Gln＋Ala209Glu＋Ser210AspGly155Asn＋Ser160Asp＋Tyr161Glu＋Asn198Ser＋Leu211SerSer126Glu＋Pro127Glu＋Gln200Ser＋Thr207Ser＋Leu211ProAla131Gly＋Ala209Thr＋Ser210Asp＋Leu211Glu＋Thr214GlyGly 61Asn＋Ile159Ser＋Ser206Glu＋Thr207Asp＋Leu211AlaGly 98Glu＋Ser 99Glu＋Tyr161Leu＋Phe183Gln＋Asn198GlnGln 57Ser＋Val 93Asn＋Gly155Asn＋Gln200Glu＋Ser210GluThr 64Asn＋Gly 95Glu＋Ala 96Asp＋Gly 98Asp＋Gln200SerIle105Ser＋Ser210Glu＋Leu211Asp＋Asn212Asp＋Gly213SerAla 96Asn＋Gly100Ser＋Ile105Cys＋Asn198Asp＋Ser210GluAla 96Gln＋Gly 98Asn＋Asn198Asp＋Pro204Asn＋Ser210AspGly 98Gln＋Asn198Glu＋Gln200Glu＋Tyr208Asn＋Ser210AspIle105Pro＋Leu124Asn＋Ile159Ser＋Asn198Asp＋Leu211AspLeu 94Gln＋Tyr161Pro＋Asn198Glu＋Thr207Gly＋Leu211GluTyr161Gly＋Val197Ala＋Asn198Gln＋Pro204Glu＋Ser206AspLeu124Ala＋Asn198Asp＋Tyr208Gly＋Ser210Glu＋Asn212AspGln185Asn＋Asn198Asp＋Gln200Glu＋Thr207Gln＋Leu211AspPro127Gly＋Tyr161Ala＋Pro204Glu＋Thr207Asp＋Thr214SerGln 57Asn＋Leu124Ser＋Val197Glu＋Asn198Glu＋Gln200GluAsp 58Glu＋Ala 96Asp＋Val197Ala＋Gly205Ser＋Thr207GlySer158Asp＋Ser160Asp＋Pro195Ser＋Thr207Ser＋Leu211MetAsp 58Glu＋Asn 60Glu＋Thr 64Glu＋Gly157Asn＋Leu211AlaLeu124Glu＋Ser126Asp＋Ile159Met＋Phe183Val＋Val193HisLeu 94Ile＋Pro127Glu＋Pro129Asp＋Ser130Asp＋Tyr203HisLeu 94Cys＋Ser 99Asp＋Ser101Glu＋Asn198Gln＋Asn212GlnGly 61Asn＋Ser128Glu＋Ser160Glu＋Ala181Asn＋Thr207GlnTyr161Met＋Val197Asp＋Gln200Asp＋Ser210Glu＋Asn212GlnVal 93Ala＋Gly125Pro＋Ser154Glu＋Gly157Asp＋Ser158AspLeu 94Glu＋Ser101Asp＋Phe183Ala＋Gln200Asn＋Leu211MetLeu 94Asp＋Ser103Glu＋Ser104Asp＋Asn198Ser＋Tyr208GlyGly125Asp＋Asn163Glu＋Ser160Glu＋Tyr161Asn＋Asn198SerLeu 94Asn＋Pro127Asn＋Ser164Asp＋Gln186Glu＋Thr214AspGly 61Gln＋Thr 64Asn＋Asn198Glu＋Ser210Asp＋Gly213AspLeu124Gln＋Asn198Glu＋Gln200Asp＋Thr207Gly＋Tyr208GluLeu 94Glu＋Leu124Asp＋Pro129Asn＋Ala166Ser＋Thr207GlyGly 98Asn＋Ala156Pro＋Asn198Glu＋Gln200Asn＋Gly213AspGly 95Glu＋Gly 98Gln＋Ser 99Glu＋Tyr102Cys＋Ser104GluGly125Asn＋Gln200Asp＋Thr207Glu＋Ser210Asp＋Leu211HisGly 59Asp＋Ser 97Asp＋Ser 99Asp＋Ala131Pro＋Pro204GlyGln 57Ser＋Tyr102Gln＋Asn198Glu＋Ala209Glu＋Leu211SerAsn153Asp＋Ile159Asp＋Tyr161His＋Phe183His＋Gly213ProGly100Ser＋Gly155Asp＋Ile159Glu＋Val197Met＋Thr207ProPro127Ser＋Gln200Asp＋Thr207Gly＋Ser210Asp＋Gly213GluAla 96Thr＋Ser126Glu＋Gly152Glu＋Ser158Glu＋Thr207AsnLeu 94Ala＋Gly 95Asp＋Ser 97Asp＋Ser101Asp＋Thr202GlyGly152Ser＋Gly205Ser＋Thr207Glu＋Ser210Glu＋Leu211AspTyr161Pro＋Pro195Ser＋Thr207Asp＋Ser210Glu＋Leu211AspAsp 58Glu＋Ser 97Glu＋Ser 99Asp＋Gln185Asn＋Gly196SerThr 64Gly＋Ser101Glu＋Ile105Glu＋Ser126Asp＋Tyr161LeuThr 64Gly＋Ser 99Asp＋Ile105Glu＋Gly205Pro＋Leu211AlaAsn153Glu＋Ala194Thr＋Asn198Glu＋Tyr208Ile＋Gly213GluGly152Gln＋Gln200Asn＋Ser206Glu＋Ala209Asp＋Ser210GluSer103Glu＋Gly125Pro＋Ala131Glu＋Tyr161Cys＋Leu211ThrGly100Ser＋Gln200Asn＋Thr207Glu＋Ser210Asp＋Leu211GlnGly100Pro＋Asn198Asp＋Gln200Glu＋Gly205Gln＋Thr207GluAla 96Asp＋Ser 99Glu＋Ser104Asp＋Val197Met＋Gly213SerGly 98Asp＋Gly100Pro＋Ser101Asp＋Gly125Ser＋A1a156SerGly 98Asp＋Gly125Asp＋Ser160Glu＋Tyr161Ile＋Thr214GlyGly 61Glu＋Gly155Gln＋Asn198Gln＋Ser206Glu＋Tyr208HisAsp 58Glu＋Asn 60Glu＋Leu124Asn＋Ser206Asp＋Thr207AsnGly 61Asp＋Thr202Pro＋Thr207Gln＋Ser210Glu＋Leu211ProGly 61Glu＋Tyr102His＋Phe183His＋Gln185Asn＋Ser210AspSer 99Asp＋Ser126Asp＋Pro129Asp＋Gly152Ser＋Leu211HisLeu 94Ile＋Gly125Gln＋Ser128Asp＋Pro129Glu＋Ser154AspAsn198Asp＋Gln200Ser＋Gly205Pro＋Gly213Ser＋Thr214AspGly 61Asn＋Ser182Asp＋Val193Gln＋Asn198Asp＋Ser210GluGly 61Glu＋Leu 94Cys＋Gln200Ser＋Gly205Glu＋Ser210GluAsp 58Glu＋Phe183Ser＋Pro204Glu＋Thr207Ser＋Ser210GluLeu 94Met＋Ala156Asp＋Tyr161Cys＋Asn212Glu＋Thr214GluAsp 58Glu＋Leu124Pro＋Ile159Thr＋Leu211Asp＋Asn212GluGln 57Glu＋Leu 94Thr＋Val197Ser＋Ala209Glu＋Ser210AspSer104Glu＋Pro129Ser＋Ala156Gly＋Ala209Glu＋Ser210GluAsn153Asp＋Ser154Glu＋Asn198Asp＋Thr207Gln＋Leu211GlnGly 59Asp＋Asn 60Glu＋Ser101Glu＋Tyr102His＋Thr207AsnGly 61Gln＋Ser104Asp＋Ile105Asp＋Asn153Ser＋Thr207AspSer 97Glu＋Val197Gln＋Asn198Gln＋Asn212Asp＋Gly213AspTyr102Val＋Ser154Glu＋Gly205Glu＋Ser206Asp＋Leu211CysAsn153Glu＋Tyr161Thr＋Pro195Gly＋Ser206Asp＋Thr207GluGly 59Gln＋Thr 64Pro＋Gly 98Glu＋Ser130Asp＋Ala131GluGly125Pro＋Gly152Asn＋Ser158Glu＋Ile159Glu＋Ser210GluSer 97Asp＋Gly 98Asp＋Ile105Val＋Leu124Thr＋Gln200AspGly100Glu＋Ser101Asp＋Ser160Asp＋Val197Ala＋Leu211GlnSer103Asp＋Ser104Asp＋Ala181Ser＋Asn198Asp＋Tyr208IleSer 99Glu＋Gly100Glu＋Gly125Asn＋Ser182Asp＋Thr207GlyLeu 94Gly＋Gly100Asp＋Ala131Gly＋Gln200Glu＋Ser210AspSer 97Glu＋Ala131Gln＋Gln200Asp＋Thr207Gly＋Ser210GluGly125Asn＋Ser130Glu＋Phe183Asn＋Gln200Asp＋Ser210GluGly 95Gln＋Tyr102Leu＋Pro127Glu＋Gln200Asp＋Ser210AspGly125Glu＋Gly155Asn＋Gln200Glu＋Ser210Asp＋Thr214GlyGln 57Ser＋Asp 58Glu＋Gln200Asp＋Thr207Gly＋Ser210GluPro127Asp＋Tyr161Cys＋Gln185Asn＋Gln200Asp＋Ser210AspGly100Glu＋Gln200Asp＋Thr207Pro＋Ser210Asp＋Leu211AlaGly 59Pro＋Ser160Asp＋Asn198Gln＋Gln200Asp＋Ser210GluThr 64Glu＋Ile105Cys＋Leu124Ser＋Val197Glu＋Ser210AspGln 57Glu＋Val 93Glu＋Gly 95Ser＋Ser103Glu＋Asn198GlnAsn 60Glu＋Ala 96Asp＋Tyr102Glu＋Gln200Ser＋Thr207GlyGln 57Ser＋Asn 60Ser＋Leu 94Gly＋Gln200Glu＋Gly213AspAsn198Asp＋Pro204Gln＋Ser206Glu＋Ser210Glu＋Leu211ThrVal 93Glu＋Ser126Glu＋Ser160Asp＋Val197Gln＋Leu211SerGly 98Asp＋Ser126Glu＋Pro129Asp＋Ala181Ser＋Gly213SerAla 96Glu＋Gly 98Glu＋Ser103Asp＋Pro127Ser＋Ile159ProGly 63Gln＋Ser128Glu＋Gly155Ser＋Asn198Glu＋Ser210GluGln 57Asp＋Ala 96Thr＋Gln185Asn＋Asn198Asp＋Ser210AspAsn 60Gln＋Gly 95Asp＋Asn198Glu＋Gln200Asn＋Ser210AspSer104Asp＋Ile105Met＋Asn198Asp＋Ser210Asp＋Leu211AsnTyr102Glu＋Asn198Asp＋Gln200Asn＋Ser210Asp＋Leu211ValGln 57Asp＋Gly125Ser＋Asn198Asp＋Gly205Asn＋Ser210AspTyr102Leu＋Gln185Glu＋Asn198Glu＋Gln200Ser＋Ser210GluGly 59Asp＋Tyr102Ile＋Asn198Glu＋Ser210Asp＋Thr214SerVal 93Thr＋Leu124Val＋Ile159Glu＋Asn198Asp＋Ser210GluSer101Asp＋Pro129Gly＋Asn198Glu＋Thr207Ser＋Leu211GluGly152Gln＋Gly155Gln＋Asn198Asp＋Gly205Asp＋Leu211AspGly 59Gln＋Thr 64Ser＋Ala156Asp＋Ser158Glu＋Gly205GluPro127Glu＋Asn198Ser＋Pro204Glu＋Ser206Asp＋Thr207ProAsp 58Glu＋Ile159Gly＋Asn198Asp＋Leu211Ile＋Asn212GluSer130Asp＋Ile159Gly＋Asn198Glu＋Leu211Met＋Asn212GluAsp 58Glu＋Asn 60Glu＋I1e159Met＋Tyr161Gln＋Gln200AspSer 97Asp＋Ser103Glu＋Ile105Glu＋Val197Cys＋Leu211ValSer103Glu＋Ile105Glu＋Asn153Ser＋Gln200Ser＋Ser210AspAla 96Pro＋Gly152Asp＋Ser160Asp＋Thr207Glu＋Leu211GlnGly 95Asp＋Ser 99Glu＋Ile105Gly＋Gly157Pro＋Leu211GluSer 97Asp＋Ser 99Glu＋Val197Glu＋Asn198Ser＋Thr214GlyGln 57Asn＋Ser 97Glu＋Ser 99Glu＋Asn153Glu＋Val193HisThr 64Glu＋Ala131Asn＋Tyr208Leu＋Ser210Asp＋Gly213SerGln 57Glu＋Ala156Pro＋Ser158Asp＋Gln185Glu＋Thr207GlnSer 97Asp＋Pro129Asn＋Ser158Glu＋Gln185Glu＋Leu211SerAla 96Thr＋Ala156His＋Ser158Glu＋Gln185Asp＋Leu211AspLeu 94Ala＋Ser101Asp＋Ser126Glu＋Ala131Gln＋Ser154AspGly157Glu＋Ile159Leu＋Gln200Asn＋Pro204Asp＋Thr207GluLeu 94Thr＋Ser126Glu＋Pro204Glu＋Thr207Asp＋Gly213AsnGly100Gln＋Ser154Glu＋Asn198Glu＋Gln200Asp＋Leu211SerGln 57Glu＋Asn198Glu＋Gln200Glu＋Tyr203Val＋Gly213ProIle105Met＋Ser158Glu＋Ile159Ser＋Asn198Glu＋Gln200AspSer104Asp＋Alal81Gln＋Asn198Glu＋Gln200Asp＋Thr207GlyGly100Asn＋Gly152Pro＋Ser182Asp＋Gln200Asp＋Gly213AspTyr102Met＋Ser128Glu＋Ser130Asp＋Asn198Asp＋Leu211MetSer128Glu＋Ser130Glu＋Ala131Thr＋Asn198Glu＋Asn212GlnVal 93Thr＋Leu124Glu＋Ile159Asn＋Gln200Glu＋Leu211AspGly157Ser＋Ser182Asp＋Gln200Glu＋Thr207Pro＋Leu211GluSer130Glu＋Gln200Asp＋Gly205Gln＋Thr207Gln＋Leu211GluGln 57Asn＋Gly 59Asn＋Asn 60Glu＋Gly 98Glu＋Ser130AspGly 61Asp＋Tyr102Pro＋Ser206Glu＋Thr207Asn＋Ser210GluThr 64Asn＋Ser 97Asp＋Pro129Asn＋Val197Asp＋Leu211GluSer 99Glu＋Ala181Thr＋Val197Glu＋Ala209Asn＋Leu211AspGly 59Asp＋Val 93Ser＋Asn153Ser＋Val197Glu＋Leu211AspSer101Glu＋Asn153Ser＋Tyr208Met＋Leu211Glu＋Gly213AspGly157Asp＋Gln185Asn＋Thr207Glu＋Ala209Asp＋Asn212GlnGly100Gln＋Gly125Ser＋Phe183Asp＋Ser210Asp＋Thr214GluAsn198Gln＋Ser206Glu＋Thr207Pro＋Ser210Asp＋Asn212AspAla181Thr＋Gln200Glu＋Thr207Asp＋Leu211Ala＋Gly213GluAsn 60Asp＋Gly 98Pro＋Ser126Glu＋Pro129Glu＋Gln200SerAsp 58Glu＋Ile159Pro＋Ser210Glu＋Asn212Asp＋Thr214GlnAsp 58Glu＋Gly125Ser＋Ala131Ser＋Ser210Asp＋Asn212GluGly 59Ser＋Ser101Glu＋Ala131His＋Ser210Glu＋Asn212GluAsn 60Gln＋Val 93Ala＋Ser103Glu＋Ser210Glu＋Asn212GluSer158Glu＋Asn198Ser＋Tyr208Glu＋Ser210Asp＋Leu211SerAla 96Ser＋Ser160Asp＋Asn198Glu＋Asn212Ser＋Thr214GluLeu 94Asn＋Gly 98Asp＋Gly100Glu＋Pro127Glu＋Val197MetAsn 60Ser＋Gly 98Asp＋Gly100Asp＋Tyr102Asn＋Gln200GluLeu 94Ser＋Ala 96Thr＋Ile105Asp＋Tyr161Glu＋Leu211IleGly 95Ser＋Ser 99Asp＋Ser101Asp＋Leu211Met＋Gly213GluGly 61Glu＋Gly125Ser＋Ser128Glu＋Ser160Asp＋Ala181ProSer182Glu＋Val193Met＋Asn198Glu＋Gly205Gln＋Ala209AspLeu 94Glu＋Gly100Asn＋Ser104Asp＋Ile105Val＋Ser206GluThr 64Asp＋Ser126Asp＋Gly152Asp＋Pro204Gly＋Leu211AsnGly 59Asp＋Val 93Glu＋Leu 94Pro＋Ile105Met＋Leu211SerPro129Glu＋Ser160Asp＋Gln200Ser＋Gly205Gln＋Ser210GluIle159Val＋Ser160Glu＋Phe183Glu＋Pro204Gln＋Ala209AsnLeu 94Asp＋Val197Asp＋Thr207Gly＋Ser210Asp＋Leu211MetLeu124Glu＋Phe183Pro＋Val197Glu＋Tyr208Leu＋Ser210GluLeu 94Met＋Leu124Asp＋Asn153Asp＋Tyr161Ala＋Leu211GluGly 98Asp＋Gly152Glu＋Ala156Asp＋Tyr208Met＋Asn212SerVal 93Asp＋Gly 98Gln＋Ser126Asp＋Ser130Glu＋Ala209ProAsp 58Glu＋Tyr102Ala＋Ser126Asp＋Ser130Glu＋Ile159AsnSer126Asp＋Ser130Glu＋Val197Gly＋Leu211Glu＋Asn212SerSer104Glu＋Ile105Pro＋Asn198Asp＋Leu211His＋Gly213AspSer 97Asp＋Gly100Glu＋Gly152Pro＋Tyr161Glu＋Ala181GlnSer128Glu＋Ala131Glu＋Ser154Asp＋Asn198Gln＋Ala209HisSer 99Asp＋Pro129Ser＋Ser154Glu＋Ser160Asp＋Pro204GlnAsp 58Glu＋Ile105Cys＋Leu124Ser＋Ser154Glu＋Ser158GluAla131Asp＋Val197Glu＋Gln200Glu＋Thr207Pro＋Thr214GlySer 99Asp＋Val197Asp＋Asn198Gln＋Gln200Glu＋Thr207GlySer101Glu＋Val197Asp＋Gln200Glu＋Pro204Gly＋Thr207SerAla131Asp＋Val197Asp＋Asn198Gln＋Gln200Glu＋Ala209AsnGly 95Pro＋Gly 98Asp＋Leu124Glu＋Thr207Asn＋Asn212GluGln 57Glu＋Asn 60Glu＋Val 93Thr＋Gly 95Asn＋Ser182AspAsn 60Asp＋Ile105Gln＋Ala131Thr＋Thr207Ser＋Ala209GluSer103Glu＋Gln200Glu＋Thr207Pro＋Ala209Gln＋Asn212AspGly 61Pro＋Leu 94Glu＋Gln200Asp＋Leu211Asn＋Asn212GluGly 59Pro＋Val 93Glu＋Gln200Asp＋Leu211Val＋Asn212AspSer 97Glu＋Gly157Ser＋Ala181Pro＋Gln200Glu＋Asn212GluVal 93Thr＋Gly 98Gln＋Tyr102Glu＋Ala131Glu＋Ser210AspGly 61Glu＋Ser104Asp＋Phe183Ala＋Thr207Gly＋Ala209GluAla156Asp＋Ser160Asp＋Ala181Gly＋Asn198Ser＋Gln200AspGln 57Glu＋Gly 61Asp＋Asn153Gln＋Asn198Asp＋Gly205SerGly100Glu＋Ile105Ser＋Leu124Glu＋Val197Met＋Ser210AspTyr102Glu＋Ser126Glu＋Val193Pro＋Pro204Asn＋Ser210AspGly 59Pro＋Ala131Asp＋Asn153Ser＋Ser154Asp＋Thr214AspSer103Glu＋Ser130Glu＋Gln200Ser＋Gly205Glu＋Thr214GlyAsp 58Glu＋Ala156Pro＋Gly157Asp＋Ser160Asp＋Gln200SerThr 64Glu＋Leu 94Asp＋Ala 96Gln＋Gln185Ser＋Ser210AspSer130Asp＋Gly157Glu＋Phe183Glu＋Asn198Gln＋Leu211MetAsp 58Glu＋Pro129Glu＋Ala156His＋Pro204Ser＋Gly205GluSer126Glu＋Ile159Leu＋Ala209Asp＋Leu211Ala＋Asn212GluGly 59Asp＋Ser126Asp＋Ser154Glu＋Phe183Leu＋Leu211ValGly155Asn＋Gly157Asp＋Gln200Glu＋Thr207Glu＋Leu211AsnGly 59Glu＋Gly 95Pro＋Ala181His＋Gln200Glu＋Thr207AspGln 57Glu＋Pro127Gly＋Gly152Ser＋Gln200Asp＋Thr207AspAla 96Glu＋Asn198Ser＋Gln200Glu＋Thr207Glu＋Thr214SerThr 64Asp＋Leu124Ser＋Pro129Asp＋Tyr161Asn＋Thr207AspSer 97Asp＋Tyr161Gly＋Ser210Glu＋Leu211Pro＋Gly213AspPro127Ser＋Pro129Asn＋Gly157Asp＋Ser210Asp＋Gly213GluTyr102Cys＋Ser130Asp＋Pro204Gln＋Ser210Glu＋G1y213AspSer 99Glu＋Ala156Ser＋Ala209Thr＋Ser210Asp＋Gly213AspGly196Pro＋Gln200Asp＋Pro204Asp＋Thr207Gln＋Leu211AsnAsp 58Glu＋Ser 97Glu＋Ser104Asp＋Pro129Gly＋Tyr161ThrAsp 58Glu＋Leu 94Ser＋Ser 97Asp＋Gly 98Pro＋Gln200AspGly 61Glu＋Ser101Asp＋Thr207Asp＋Leu211Ile＋Thr214SerGly 61Asp＋Ser101Glu＋Tyr102Val＋Thr207Glu＋Thr214SerGly 61Asp＋Asn153Gln＋Ile159Gly＋Asn198Asp＋Thr207GluGly 61Glu＋Ile159Ser＋Val197Met＋Asn198Glu＋Thr207AspThr 64Gln＋Gly100Glu＋Gln200Asn＋Thr207Glu＋Ser210AspSer126Glu＋Gly155Pro＋Tyr161Thr＋Thr207Glu＋Ser210AspTyr102Glu＋Gly125Gln＋Ala181Pro＋Thr207Asp＋Ser210GluGly100Glu＋Gly205Ser＋Thr207Glu＋Ser210Glu＋Asn212GlnGly 59Ser＋Gly 95Glu＋Thr207Glu＋Ser210Asp＋Leu211ValTyr161Asp＋Asn198Ser＋Thr207Glu＋Ser210Glu＋Gly213ProPro129Gln＋Ser130Glu＋Asn198Gln＋Thr207Asp＋Ser210GluGln 57Asp＋Phe183Gly＋Pro204Gln＋Thr207Glu＋Ser210AspAsn 60Gln＋Gly157Glu＋Pro204Gly＋Thr207Asp＋Ser210AspPro129Asp＋Gly152Pro＋Thr207Asp＋Ser210Glu＋Leu211AsnGly 61Glu＋Ser 99Asp＋Gly155Asn＋Pro195Gln＋Gln200AsnGly 98Glu＋Ser101Glu＋Ala181Gly＋Gln185Glu＋Thr207ProGly 59Pro＋Thr 64Asp＋Pro129Gln＋Ala181Glu＋Asn198AspGly 98Asp＋Tyr161Met＋Ala181Glu＋Asn198Asp＋Leu211ValIle159Glu＋Val197Thr＋Tyr208Gln＋Ala209Glu＋Thr214AspSer126Glu＋Gly157Glu＋Ser182Glu＋Asn198Gln＋Leu211ValGly 61Asp＋Ser128Glu＋Ser206Asp＋Tyr208Gly＋Leu211MetGly 98Pro＋Ser130Glu＋Ser160Glu＋Thr207Gly＋Ser210AspGln 57Asp＋Gly125Gln＋Ser130Asp＋Asn153Ser＋Ser160AspTyr161Met＋Gln200Glu＋Ser206Glu＋Ala209His＋Gly213AspGly 61Asp＋Ala156Glu＋Gly205Asn＋Thr207Gln＋Ser210GluIle105Glu＋Ser130Asp＋Gln200Asn＋Gly205Glu＋Thr207AsnGln 57Ser＋Asn 60Asp＋Gln200Asp＋Gly205Glu＋Thr207SerSer101Glu＋Ser126Glu＋Val197Asp＋Thr207Gly＋Leu211CysSer101Asp＋Ser126Asp＋Gly152Ser＋Asn19SGlu＋Gln200AsnGln 57Asp＋Gly125Asp＋Gly205Asp＋Thr207Ser＋Tyr208ThrLeu 94Met＋Ser123Asp＋Ser158Glu＋Asn198Gln＋Ser210AspGly 59Asp＋Gly 95Pro＋Ala194Gln＋Asn198Asp＋Thr214AspGln 57Glu＋Gly 61Asn＋Gly 98Asp＋Gly100Pro＋Val193CysThr 64Gln＋Val 93Glu＋Ser 97Glu＋Pro129Ser＋Gln200GluGln 57Glu＋Thr 64Asn＋Val 93Asp＋Gln200Glu＋Thr207AsnGly100Asp＋Gly125Ser＋Ser126Glu＋Gly196Gln＋Val197GluVa1 93Met＋Gly100Asp＋Ser126Glu＋Tyr161Ala＋Asn198AspGly100Glu＋Ser126Asp＋Ala181His＋Val197Cys＋Gln200GluAsn 60Gln＋Ser 99Asp＋Ser103Glu＋Ser210Asp＋Leu211AlaGln 57Ser＋Ser 99Glu＋Ser103Glu＋Gln200Asp＋Thr207AsnGly 63Ser＋Ala 96Ser＋Ser210Glu＋Leu211Pro＋Thr214AspPro129Gly＋Val197Gln＋Ser210Glu＋Leu211His＋Thr214AspPro127Asp＋Gly155Glu＋Tyr161Ser＋Thr207Ser＋Ser210GluAsp 58Glu＋Val 93His＋Gly155Pro＋Gln200Asn＋Thr207GluThr 64Asn＋Asn198Ser＋Gln200Glu＋Ser206Glu＋Thr214GlnGly 59Pro＋Gly 61Glu＋Ser154Asp＋Gln200Asp＋Thr214AsnSer126Asp＋Pro129Ser＋Ala156Asp＋Ser210Asp＋Leu211AlaSer103Glu＋Ile159Leu＋Pro204Glu＋Gly205Asn＋Ser210AspAla131Thr＋Asn153Glu＋Tyr203Asn＋Pro204Glu＋Ser210GluGly157Asn＋Ser160Glu＋Pro204Glu＋Gly205Ser＋Ser210GluGly 95Gln＋Gly 98Pro＋Gly152Glu＋Gly157Pro＋Val197AspGln 57Glu＋Leu 94Pro＋Gly125Ser＋Asn153Glu＋Leu211GluGly 59Ser＋Ser104Asp＋Ser130Glu＋Ser210Glu＋Leu211SerSer104Glu＋Ser130Asp＋Gly155Asn＋Leu211Gly＋Gly213AspSer104Glu＋Ser130Glu＋Asn198Asp＋Gln200Asn＋Thr207GlnSer158Glu＋Thr202Ser＋Thr207Pro＋Leu211Thr＋Thr214AspGly 59Glu＋Leu 94Val＋Ser101Asp＋Ala131Glu＋Tyr161MetAsp 58Glu＋Ser 99Glu＋Tyr102Asn＋Ala209Asn＋Ser210GluSer101Asp＋Phe183Ser＋Gln200Asp＋Pro204Glu＋Thr214GlnGly 61Asp＋Gly 98Asn＋Ser 99Glu＋Ile105Ser＋Val197GluAsn 60Ser＋Gly125Pro＋Ala156Asp＋Tyr161Asp＋Ser210AspGly 59Pro＋Ser128Glu＋Ser154Asp＋Thr207Glu＋Leu211IleSer103Asp＋Ala156Gly＋Ser182Glu＋Asn198Glu＋Tyr203HisVal 93Ala＋Gly125Pro＋Ser160Glu＋Thr207Asp＋Leu211GluSer 97Asp＋Gly152Pro＋Thr207Glu＋Leu211Glu＋Gly213GlnGln 57Glu＋Ser103Asp＋Pro129Glu＋Ala181Gly＋Thr207ProSer158Asp＋Tyr161Pro＋Ser182Asp＋Thr207Gln＋Ser210Asp表37多个环状区的六突变的突变体Tyr102Cys＋Ile105Val＋Leu124Ile＋Ser154Glu＋Asn198Gln＋Thr207GlySer160Glu＋Tyr203Cys＋Thr207Pro＋Leu211Met＋Asn212Gln＋Gly213GlnVal 93Ser＋Ser103Glu＋Gly125Ser＋Gln185Asn＋Ala194Gln＋ Asn198GlnGly100Ser＋Ile105Met＋Gly157Pro＋Gln200Asn＋Thr207Asn＋Leu211ValThr 64Glu＋Leu 94Asn＋Val197Cys＋Thr202Gln＋Thr207Gln＋Leu211ValGly152Pro＋Gly157Gln＋Ala181Asp＋Asn198Gln＋Gln200Asn＋Thr207AsnGly 95Gln＋Ala 96Asn＋Leu124Thr＋Asn153Ser＋Ser154Glu＋Val197HisVal 93Met＋Pro127Ser＋Gly155Pro＋Ser182Asp＋Gln200Ser＋Gly213SerGly 61Pro＋Gly 95Pro＋Ser154Glu＋Ala181His＋Leu211Asn＋Gly213ProTyr161Thr＋Pro195Asn＋Gln200Asn＋Thr207Gly＋Tyr208Leu＋Leu211ProLeu 94Pro＋Ala 96Ser＋Pro127Asp＋Ile159Ala＋Thr207Pro＋Gly213GlnGly100Asn＋Ile105Gln＋Gly125Gln＋Pro129Gly＋Gln185Glu＋Thr207GlnGly 98Glu＋Pro129Gly＋Gly155Pro＋Thr207Gly＋Ala209His＋Thr214GlyIle105Gln＋Gly152Asp＋Phe183His＋Ala194Gln＋Gly205Ser＋Asn212GlnAla194Gln＋Asn198Ser＋Gln200Asn＋Ser206Asp＋Leu211His＋Thr214GlnSer 99Glu＋Pro129Asn＋Ala131Pro＋Gln185Asn＋Thr207Pro＋Leu211ValLeu 94Val＋Gly 98Gln＋Leu124Gly＋Val197Glu＋Gln200Asn＋Thr207GlyGln 57Ser＋Gly 98Asn＋Tyr102Asp＋Tyr161Asn＋Gln200Ser＋Thr207GlnAsn 60Gln＋Gly 61Ser＋Tyr102Cys＋Gly152Asp＋Val199Ser＋Pro204AsnTyr102Thr＋Gly155Glu＋Ala156Asn＋Tyr161Pro＋Gln200Asn＋Leu211ThrTyr102Glu＋Gly157Pro＋Phe183His＋Asn198Gln＋Thr207Pro＋Leu211CysLeu124Cys＋Pro127Asn＋Tyr208Met＋Ser210Asp＋Leu211His＋Gly213GlnGly 98Ser＋Leu124Ala＋Ser126Asp＋Ala181Asn＋Ala209Gly＋Gly213GlnAla 96His＋Tyr102Ser＋Ile105Ser＋Gly157Ser＋Ile159Gln＋Leu211AspThr 64Pro＋Ala 96Pro＋Ser 97Asp＋Pro127Ser＋Gly205Asn＋Leu211SerVal 93Gly＋Gly 98Ser＋Tyr102Asn＋Val197Met＋Thr207Gln＋Ser210AspAsn 60Gln＋Ala 96Gln＋Asn153Glu＋Thr202Gln＋Gly205Asn＋Leu211MetGly 59Asn＋Leu 94Val＋Pro127Ser＋Ile159Ala＋Gln200Asn＋Leu211MetIle105Gln＋Gly157Gln＋Val193Asn＋Val197Asp＋Thr207Pro＋Tyr208CysAsn 60Gln＋Gly 63Gln＋Ile105Glu＋Asn198Gln＋Leu211Ile＋Thr214GlyGly 61Gln＋Gly125Glu＋Tyr161Gly＋Gly205Gln＋Thr207Ser＋Ala209GlnGly 98Ser＋Ala156Gln＋Tyr161Ala＋Ser182Glu＋Gln200Ser＋Thr207GlnGly 61Pro＋Val 93Ala＋Leu124Gly＋Ala131Thr＋Val197Glu＋Asn198GlnVal 93Asp＋Ala 96Asn＋Val197His＋Asn196Ser＋Val199Ser＋Thr207GlnTyr102Ile＋Ile159Asp＋Ala181Asn＋Gln200Ser＋Pro204Gln＋Leu211HisGly100Prp＋Pro129Ser＋Gln200Ser＋Thr207Pro＋Ala209Glu＋Ser210GluVal197Pro＋Asn198Gln＋Gln200Glu＋Pro204Gly＋Thr207Gln＋Ser210AspAla 96Gln＋Leu124Met＋Ile159Ser＋Gln200Glu＋Pro204Gly＋Ser210AspAsn 60Gln＋Ile159Thr＋Asn198Ser＋Gln200Glu＋Ser210Glu＋Leu211AspGly 61Ser＋Val197Gln＋Asn198Glu＋Gln200Ser＋Ser210Asp＋Leu211HisGly100Asn＋Ala181Pro＋Asn198Asp＋Thr207Ser＋Ser210Glu＋Gly213GlnGly 98Asn＋Pro129Ser＋Asn198Asp＋Gln200Asp＋Ser210Glu＋Leu211ProGly 63Ser＋Leu 94Gln＋Ile159Ala＋Gly205Glu＋Thr207Glu＋Thr214AsnAsp 58Glu＋Gly 61Glu＋Gly 98Asn＋Asn153Ser＋Tyr161Asn＋Leu211CysLeu124Ser＋Ile159Thr＋Asn198Glu＋Gln200Glu＋Pro204Gly＋Thr207GlnGly 61Gln＋Gly 95Asn＋Ala194Gly＋Asn198Asp＋Gln200Asp＋Leu211ValGln 57Ser＋Tyr102Gln＋Asn198Glu＋Ala209Glu＋Ser210Glu＋Leu211SerGly 59Gln＋Gly125Pro＋Ser154Glu＋Gly155Asp＋Tyr161Ala＋Ser182AspGln 57Glu＋Gly 59Asp＋Thr 64Gln＋Tyr208Pro＋Ala209Gln＋Leu211AsnAsn 60Asp＋Gly 61Gln＋Gly 95Glu＋Gly 98Glu＋Asn153Ser＋Ile159LeuIle105Thr＋Gly152Ser＋Ser158Asp＋Ser160Asp＋Thr207Gly＋Leu211AlaGly 61Gln＋Thr 64Asn＋Asn198Glu＋Ser210Asp＋Leu211Asp＋Gly213AspLeu 94Pro＋Ala 96Gly＋Thr207Ser＋Ser210Asp＋Leu211Cys＋Asn212GluGly 98Glu＋Gly100Asp＋Ala181Pro＋Tyr203Val＋Pro204Asn＋Thr207SerAsp 58Glu＋Gly157Ser＋Gln185Asn＋Val197Gln＋Pro204Glu＋Asn212GlnGly 61Asn＋Leu124Thr＋Ser128Glu＋Ser160Glu＋Ala181Asn＋Thr207GlnIle105Leu＋Leu124Ser＋Gly125Asp＋Gly152Glu＋Asn153Glu＋Gly213GluLeu124Gln＋Asn198Glu＋Gln200Asp＋Thr207Gly＋Tyr208Glu＋Leu211AspLeu 94Ile＋Gly152Ser＋Gly15SGlu＋Ser158Asp＋Ala181Gly＋Val197HisAsp 58Glu＋Tyr161Gly＋Val197Ala＋Asn198Gln＋Pro204Glu＋Ser206AspGln 57Ser＋Ile159Ser＋Asn198Asp＋Gln200Asn＋Tyr208Asn＋Gly213GluGly152Ser＋Phe183Ala＋Asn198Glu＋Val199Ser＋Leu211Cys＋Gly213GluLeu 94Ile＋Ser154Asp＋Ile159Glu＋Asn198Ser＋Gln200Asn＋Leu211MetAla 96Pro＋Ser154Asp＋Ala156His＋Gly157Asn＋Ser158Glu＋Pro204SerGln 57Asp＋Gly 59Pro＋Asn 60Asp＋Gly 95Pro＋Thr207Ser＋Ala209GlnThr 64Ser＋Gly152Gln＋Ala181His＋Gln200Glu＋Leu211His＋Asn212GluGln 57Asn＋Gly 98Gln＋Gln200Asp＋Leu211Cys＋Asn212Asp＋Gly213SerAla 96Asn＋Gly100Asp＋Leu124Asp＋Val197Gly＋Thr207Pro＋Gly213SerAla 96Ser＋Ser154Asp＋Phe183Gln＋Thr207Ser＋Leu211Thr＋Gly213AspGly 95Asp＋Gly 98Pro＋Ser 99Asp＋Gly100Ser＋Tyr102Glu＋Ser103GluGln 57Asp＋Asp 58Glu＋Thr 64Asp＋Ser 97Glu＋Gln200Ser＋Tyr208ThrGly125Asp＋Ser130Asp＋Asn153Glu＋Ala156Thr＋Ser160Glu＋Asn198SerGly 59Asn＋Leu124Thr＋Ile159Met＋Ala209Asp＋Leu211Cys＋Asn212AspGly 95Gln＋Gly100Asn＋Ile105Met＋Ala181Gln＋Gln200Glu＋Thr207GluIle105Met＋Ser126Asp＋Asn153Glu＋Gly205Gln＋Thr207Pro＋Leu211MetPro129Gly＋Ala131His＋Val197Thr＋Thr207Asn＋Ser210Glu＋Gly213GluGly 98Asp＋Gly152Glu＋Asn153Glu＋Ser154Asp＋Val197Ser＋Gln200AsnAsp 58Glu＋Gly 61Pro＋Val 93Gly＋Ser 97Glu＋Pro129Asn＋Tyr161IleSer130Asp＋Gly152Ser＋Val197Gly＋Asn198Glu＋Ser210Glu＋Leu211AspTyr102Val＋Gly155Asn＋Ser182Asp＋Val197Asp＋Gln200Asp＋Ser210AspThr 64Gly＋Ser128Glu＋Gln200Glu＋Thr207Ser＋Ser210Asp＋Leu211AspGln 57Glu＋Tyr102Ser＋Thr207Pro＋Ser210Asp＋Leu211Asp＋Asn212AspPro127Asn＋Gly152Glu＋Ile159Asn＋Asn198Asp＋Gln200Asn＋Gly213GluAsn153Ser＋Gly155Ser＋Ile159Thr＋Asn198Asp＋Ser210Glu＋Thr214AspVal 93Thr＋Ser154Asp＋Ser158Glu＋Ser182Glu＋Asn198Ser＋ Thr207SerSer101Asp＋Gly155Pro＋Asn198Glu＋Gln200Glu＋Ala209Asn＋Ser210AspSer 99Glu＋Ser126Asp＋Pro127Asp＋Ile159Ser＋Gln200Ser＋Leu211MetPro129Asn＋Asn153Asp＋Ser158Asp＋Gln200Ser＋Pro204Ser＋Gly205SerVal197Cys＋Gly205Asp＋Thr207Asp＋Ser210Asp＋Leu211Ile＋Asn212SerGly 61Asn＋Asn198Asp＋Gln200Glu＋Thr207Glu＋Tyr208Cys＋Leu211HisAsn 60Asp＋Pro127Gln＋Asn153Glu＋Ser154Glu＋Ala156Glu＋Thr207ProSer 99Glu＋Asn153Asp＋Ser154Asp＋Ala156Glu＋Gln185Asn＋Pro195SerThr 64Asn＋Ala156Asn＋Val197Asp＋Gln200Glu＋Leu211Pro＋Thr214GluSer126Asp＋Pro127Gln＋Ser128Asp＋Ser160Asp＋Gln200Glu＋Thr214SerAsn153Asp＋Gln185Glu＋Asn198Ser＋Gln200Glu＋Leu211Cys＋Gly213AspIle105Gln＋Leu124Glu＋Gly125Glu＋Ser160Asp＋Ala194Gln＋Pro204GluAsp 58Glu＋Leu 94Pro＋Ser154Glu＋Gly155Asp＋Ser182Asp＋Asn198GlnSer 99Glu＋Val197Glu＋Thr207Gly＋Ala209Asn＋Ser210Glu＋Leu211AspGln 57Asn＋Gly125Glu＋Asn153Glu＋Gln200Asn＋Leu211Ser＋Asn212AspLeu 94Ser＋Gly125Gln＋Ser126Glu＋Val197Asp＋Ser210Asp＋Asn212GluThr 64Glu＋Ile159His＋Asn198Glu＋Gln200Asp＋Leu211Cys＋Thr214GlyGly 98Asp＋Gly152Glu＋Gly155Glu＋Ala156Asp＋Tyr208Met＋Asn212SerGly 98Pro＋Ser104Glu＋Asn198Glu＋Gln200Asp＋Gly205Gln＋Asn212AspAsp 58Glu＋Thr 64Glu＋Ala 96Ser＋Tyr102His＋Gln200Glu＋Leu211GlyLeu 94Ala＋Gly100Gln＋Phe183Asp＋Val193Gln＋Val197Asp＋Gln200GluAsn 60Gln＋Thr 64Gly＋Gly125Glu＋Ser154Glu＋Gly157Asp＋Thr207GlyVal 93Met＋Gly100Asp＋Ser126Glu＋Pro129Asp＋Ala181Asn＋Thr214SerGly125Asn＋Ser182Glu＋Phe183Tyr＋Gln185Asp＋Asn198Asp＋Asn212GluLeu 94Gln＋Gly125Asp＋Ser128Glu＋Pro129Glu＋Asn198Gln＋Ser210GluGly 59Gln＋Thr 64Asp＋Asn198Gln＋Gln200Asn＋Ser210Asp＋Asn212AspGly 59Gln＋Gly125Glu＋Asn198Asp＋Gln200Asp＋Leu211Ile＋Gly213AspThr 64Asn＋Gly125Glu＋Asn153Asp＋Gln185Glu＋Thr207Ser＋Leu211AspAsp 58Glu＋Gly 95Glu＋Gly 98Glu＋Gln185Asn＋Pro204Ser＋Ser210AspGln 57Glu＋Gly 59Glu＋Thr 64Asp＋Gly152Ser＋Ala156Glu＋Leu211SerAsn 60Ser＋Ala156Gln＋Asn198Glu＋Gly205Gln＋Thr207Asp＋Leu211AspLeu 94Ala＋Ser101Asp＋Ser154Glu＋Ser158Asp＋Ser160Asp＋Leu211ThrAsp 58Glu＋Gln200Glu＋Thr207Glu＋Ser210Glu＋Leu211Thr＋Gly213ProGly 61Gln＋Gly152Asp＋Gln200Glu＋Ser210Glu＋Leu211Gly＋Thr214AspAsn 60Glu＋Tyr102Ile＋Gln200Glu＋Thr207Asp＋Ser210Asp＋Asn212SerGly 59Ser＋Val 93Asp＋Ile159Asp＋Ser160Glu＋Thr207Asn＋Gly213SerThr 64Asn＋Ser104Asp＋Ile105Cys＋Ala156Gly＋Ser210Asp＋Leu211AspGln 57Asn＋Gly 98Asp＋Gly100Gln＋Ser206Glu＋Thr207Glu＋Tyr208MetGln 57Asn＋Ser158Asp＋Ile159Glu＋Phe183Met＋Asn198Asp＋Thr207AsnGly 95Asp＋Ala 96Gly＋Ala131Gln＋Thr207Glu＋Tyr208Glu＋Leu211ThrSer103Asp＋Ser104Asp＋Gln200Asn＋Leu211Asn＋Asn212Asp＋Gly213AspGly 61G1n＋Gly 98Glu＋Gly155Asp＋Phe183Asp＋Thr207Gln＋Leu211ProLeu 94Ser＋Gly 95Ser＋Ser 97Glu＋Gly 98Glu＋Ala131Glu＋Asn212SerThr 64Asn＋Tyr102Ile＋Ser103Asp＋Ser104Asp＋Ala181Asn＋Tyr208AspThr 64Gly＋Tyr102Asp＋Gln200Glu＋Thr207Asn＋Tyr208Gln＋Ser210GluSer 99Glu＋Tyr102His＋Ile159Ala＋Gln200Asp＋Thr207Gln＋Ser210GluThr 64Glu＋Leu124Cys＋Val197Glu＋Gln200Asn＋Ser210Asp＋Leu211GlyThr 64Gly＋Ser154Asp＋Ala181Asn＋Gln200Glu＋Thr207Glu＋Ser210AspTyr102Ala＋Ala181Asn＋Ser182Glu＋Gln200Asp＋Thr207Asp＋Ser210AspGly125Pro＋Gly152Asn＋Ser158Glu＋Ile159Glu＋Asn198Asp＋Ser210GluGln 57Ser＋Asn 60Ser＋Val 93Gln＋Leu 94Gly＋Gln200Glu＋Gly213AspGly 61Glu＋Thr 64Ser＋Val 93Asp＋Asn198Ser＋Pro204Glu＋Tyr208GlySer101Asp＋Ser103Glu＋Leu124Met＋Gly1555er＋Ser210Asp＋Leu211GlnSer 99Glu＋Pro127Asp＋Pro1295er＋Ser154Glu＋Ser158Glu＋Thr207SerGly 61Glu＋Ser101Glu＋Ser104Glu＋Ile159Leu＋Gly196Asn＋Asn198GlnGly 59Asp＋Asn 60Glu＋Ser 99Glu＋Val197Thr＋Thr207Asp＋Gly213GlnGly 61Asn＋Ala 96Glu＋Gly 98Glu＋Leu124Gln＋Pro127Ser＋Gly152GluGly125Ser＋Phe183Asp＋Gln185Glu＋Ala209Glu＋Ser210Asp＋Leu211HisSer130Asp＋Ile159Gly＋Asn198Glu＋Thr207Gln＋Ser210Asp＋Leu211MetPro127Gly＋Ser130Asp＋Tyr161Thr＋Asn198Glu＋Ser210Glu＋Thr214GlnGly 98Pro＋Tyr102Glu＋Phe183Met＋Asn198Asp＋Gln200Asn＋Ser210AspSer 99Glu＋Pro127Ser＋Gly155Pro＋Val197Cys＋Asn198Glu＋Leu211GluLeu 94Ser＋Ser101Glu＋Ile105Glu＋Pro195Gln＋Val197Glu＋Asn198GluPro127Glu＋Asn198Ser＋Pro204Glu＋Gly205Ser＋Ser206Asp＋Thr207ProGln 57Ser＋Asn 60Asp＋Gln185Asn＋Asn198Glu＋Leu211Thr＋Asn212AspAsp 58Glu＋Asn 60Asp＋Tyr102Ala＋Ile159Ser＋Ser160Glu＋Gly205ProAla 96Gly＋Ala181Asp＋Asn198Asp＋Thr207Glu＋Ala209His＋Ser210GluGln 57Asn＋Asn 60Glu＋Gly 95Glu＋Gly100Glu＋Gly125Ser＋Ser210GluAsp 58Glu＋Phe183Ser＋Pro204Glu＋Thr207Ser＋Ser210Glu＋Asn212GluLeu 94Asp＋Pro129Glu＋Ala156Ser＋Tyr161Asp＋Gln185Ser＋Asn212SerGly 95Glu＋Ser 99Asp＋Ala156His＋Val197Glu＋Asn198Gln＋Thr207AsnThr 64Glu＋Pro127Asn＋Ala156Glu＋Ala209Asp＋Ser210Asp＋Leu211AsnGly 59Ser＋Gly 63Pro＋Pro129Ser＋Gly157Glu＋Gln185Asp＋Ser210GluGln 57Ser＋Gly 59Asp＋Leu 94Glu＋Gly125Asp＋Ser160Glu＋Gln185AsnIle105Gln＋Ser126Glu＋Ser128Asp＋Leu211Glu＋Gly213Asp＋Thr214GlyPro127Ser＋Ser182Glu＋Gln185Glu＋Asn198Asp＋Thr207Pro＋Ser210GluAla 96Asn＋Ala131Pro＋Ala181Asp＋Thr207Asp＋Ser210Asp＋Leu211AspAsp 58Glu＋Gly 59Pro＋Ser126Asp＋Gly152Asp＋Ser158Glu＋Leu211IleGly155Asp＋Ser182Asp＋Val197Gly＋Gln200Asp＋Thr202Asn＋Leu211GluSer126Asp＋Ile159Ser＋Asn198Ser＋Gln200Ser＋Pro204Glu＋Thr207AspAsn 60Ser＋Gly 63Asn＋Gln185Glu＋Pro195Ser＋Asn198Asp＋Gln200GluGly 59Ser＋Ile105Asp＋Ala181Asn＋Asn198Asp＋Gln200Asp＋Leu211ThrLeu 94Glu＋Ser 99Asp＋Gly100Gln＋Ser103Asp＋Gln200Glu＋Tyr208MetAsp 58Glu＋Leu 94Asp＋Ser 99Glu＋Gly100Pro＋Gly157Asp＋Tyr203ValAsp 58Glu＋Thr 64Gln＋Gly 98Glu＋Ser 99Asp＋Pro129Glu＋Gly155SerVal 93Gln＋Ser101Asp＋Ile105Asp＋Asn198Asp＋Gly205Ser＋Leu211AsnSer 99Glu＋Ile105Ala＋Ser210Glu＋Leu211His＋Asn212Glu＋Thr214AspVal 93Ala＋Leu124His＋Ala131Gly＋Gly157Asp＋Val197Asp＋Gly213AspIle105His＋Gly152Ser＋Ala156Asn＋Ser158Glu＋Val197Asp＋Gly213AspGly 61Asp＋Leu 94Thr＋Val197Met＋Asn198Gln＋Gln200Glu＋Gly205GluTyr102Pro＋Ser128Asp＋Ser130Glu＋Ala131Thr＋Gln200Asp＋Leu211AspGly 98Ser＋Ser126Asp＋Ser128Glu＋Gly152Gln＋Ser158Asp＋Gly205GluSer130Asp＋Phe183Ile＋Thr207Gly＋Tyr208Asp＋Ser210Glu＋Asn212AspAsp 58Glu＋Leu 94Thr＋Gly100Gln＋Tyr102His＋Ser206Asp＋Thr214ProAsp 58Glu＋Thr 64Glu＋Ser126Glu＋Pro129Ser＋Ala181Gly＋Asn198GlnAsn 60Ser＋Leu 94Asp＋Ala 96Glu＋Ala181Glu＋Gln185Asn＋Thr207GlyPro127Asp＋Pro129Glu＋Ala181Gln＋Pro195Ser＋Ser206Asp＋Thr214GlnGly152Gln＋Ala156Asp＋Thr207Asp＋Ala209Glu＋Leu211Pro＋Gly213ProSer103Glu＋Gly157Ser＋Ser158Glu＋Ser160Glu＋Tyr161His＋Pro204SerLeu 94Asp＋Gly 95Gln＋Gly 98Asp＋Ser126Glu＋Ser154Asp＋Thr207SerSer 97Glu＋Ile105Glu＋Leu124Asp＋Phe183Gly＋Thr207Gly＋Leu211GlnGly100Glu＋Pro129Ser＋Ala131Ser＋Tyr161Val＋Ser210Asp＋Asn212GluGly 59Ser＋Ser101Glu＋Ile105Gln＋Ala131His＋Ser210Glu＋Asn212GluGln 57Asn＋Pro127Gln＋Ala156Asp＋Ile159Asp＋Pro195Gly＋Ser206AspVal 93His＋Gly100Glu＋Ser126Asp＋Ser128Glu＋Gln185Asp＋Pro204GlyGly 95Ser＋Ser 99Asp＋Ser101Asp＋Val197Asn＋Leu211Met＋Gly213GluGln 57Ser＋Gly 61Glu＋Ala 96Glu＋Asn153Asp＋Gly213Pro＋Thr214GluGln 57Ser＋Ala156Asp＋Ser160Asp＋Val197Gly＋Leu211Asp＋Gly213AspGly 98Pro＋Ser104Asp＋Ala181Glu＋Ser182Glu＋Asn198Glu＋ Asn212GlnIle105Glu＋Leu124Thr＋Val197Ser＋Pro204Glu＋Thr207Asp＋Ser210AspAsn 60Ser＋Gly 61Pro＋Ser160Asp＋Gln185Asp＋Gln200Asn＋Ser210GluGly 61Ser＋Leu 94Asp＋Ser104Asp＋Leu124Asn＋Asn198Ser＋Gly213AspAsn 60Glu＋Val 93Met＋Gly 95Asp＋Gly 98Asn＋Asn198Glu＋Leu211CysGly 95Ser＋Gly 98Asp＋Ser 99Glu＋Ser126Glu＋Ser182Glu＋Thr207GlyAsp 58Glu＋Gly 98Asp＋Leu124Cys＋Gln200Asp＋Ser210Asp＋Asn212SerLeu 94Glu＋Ser 97Asp＋Ser158Glu＋Val197Thr＋Tyr208Ser＋Leu211SerSer101Glu＋Ile105Glu＋Ser154Glu＋Gly157Glu＋Asn198Ser＋Leu211AlaGly 63Ser＋Gly 98Glu＋Gly100Pro＋Gly152Asp＋Gln200Asn＋Gly213AspLeu 94Glu＋Ser101Asp＋Phe183Ala＋Gln200Asn＋Ser206Glu＋Leu211MetGln 57Glu＋Asp 58Glu＋Leu124Pro＋Asn198Ser＋Ser206Asp＋Gly213AspGly125Glu＋Ala131His＋Gly205Asp＋Ala209Glu＋Ser210Glu＋Thr214SerGly 59Ser＋Gly100Glu＋Ser103Asp＋Asn198Ser＋Tyr208Leu＋Ser210GluAsp 58Glu＋Asn 60Asp＋Gly155Asp＋Val197Asn＋Thr207Ser＋Thr214AspSer 99Asp＋Ser128Glu＋Pro129Asp＋Gly152Glu＋Gln200Asn＋Thr207AsnPro127Gln＋Asn153Glu＋Ser182Asp＋Val197Cys＋Ser210Glu＋Leu211ProSer104Glu＋Leu124Pro＋Asn153Glu＋Ala156Thr＋Ile159Leu＋Ser182GluThr 64Asn＋Tyr102Cys＋Ser128Asp＋Ser158Asp＋Phe183Glu＋Asn198SerAla 96Gln＋Tyr161Asn＋Gln185Asn＋Val197Glu＋Gly205Glu＋Ser210AspGly 98Ser＋Ser101Glu＋Pro127Gly＋Gln200Asp＋Leu211Asp＋Thr214GluGly 59Gln＋Thr 64Glu＋Gly 95Ser＋Gly155Glu＋Ser158Asp＋Val197CysTyr102Val＋Ser103Glu＋Asn198Asp＋Gln200Glu＋Thr207Glu＋Ala209AsnAsn 60Glu＋Gly 98Gln＋Asn153Glu＋Ser210Asp＋Leu211Ala＋Gly213GluAsp 58Glu＋Gly 61Glu＋Ser101Glu＋Leu124Asn＋Pro129Gly＋Ser130AspSer126Glu＋Ser130Asp＋Ala131Thr＋Gln200Asn＋Thr202Gln＋Pro204AspGly 63Gln＋Ser 97Glu＋Ser126Asp＋Ser130Asp＋Asn198Ser＋Gln200SerVal 93Gln＋Leu 94Cys＋Ser 97Glu＋Ser101Glu＋Asn212Gln＋Gly213SerAsp 58Glu＋Gly 95Glu＋Ser154Asp＋Ser158Asp＋Thr207Pro＋Tyr208ValAsn 60Gln＋Thr 64Ser＋Pro129Asn＋Pro204Asp＋Ser210Asp＋Leu211IleSer 99Asp＋Ile159Gln＋Val197Gly＋Asn198Glu＋Lau211Val＋Gly213AspSer 97Glu＋Gly100Glu＋Gly152Glu＋Gly155Gln＋Ala181Ser＋Ala209HisAsp 58Glu＋Ala 96Glu＋Ser126Glu＋Asn153Asp＋Phe183Leu＋Gly205AsnGly 98Asp＋Pro127Glu＋Gly152Pro＋Ser158Glu＋Gly205Pro＋Leu211GlnSer154Glu＋Gly157Asp＋Ile159Asn＋Asn198Glu＋Gln200Asn＋Tyr203AsnGly 63Ser＋Phe183Glu＋Thr207Glu＋Ser210Glu＋Leu211pro＋Thr214AspGly100Glu＋Asn153Ser＋Ser154Asp＋Ser160Glu＋Asn198Ser＋Gly213GluGln 57Glu＋Ala 96His＋Gly 98Pro＋Ser160Asp＋Pro204Asp＋Tyr208GlyGly 59Asp＋Gly 95Pro＋Ala194Gln＋Asn198Asp＋Gln200Asp＋Thr214AspGly 95Glu＋Val197Glu＋Asn198Gln＋Gln200Glu＋Thr207Gln＋Leu211AlaGly 98Asp＋Ser128Asp＋Gly155Pro＋Ser158Asp＋Gln185Glu＋Gln200SerGly 98Glu＋Leu124Asp＋Gly125Asn＋Ile159Gly＋Val197Asp＋Leu211AlaThr 64Asp＋Gly152Asn＋Ser160Glu＋Gln185Asn＋Thr207Asp＋Ser210AspThr 64Asp＋Gly155Asp＋Ala156Ser＋Val197Ser＋Asn198Glu＋Leu211AspGln 57Asn＋Thr 64Pro＋Ile105Glu＋Ser126Glu＋Ser160Asp＋Ser206GluGly 98Asn＋Gly157Asp＋Phe183Glu＋Val197Asn＋Ser206Asp＋Thr214GluSer 97Asp＋Ser126Glu＋Ser128Asp＋Pro129Asn＋Ala156Asp＋Leu211GlnGln 57Glu＋Leu124Pro＋Phe183Asn＋Val197Glu＋Gly205Glu＋Gly213GluSer126Asp＋Ser158Glu＋Gln200Asp＋Thr207Pro＋Leu211Glu＋Asn212SerGly 98Ser＋Gly100Pro＋Pro129Glu＋Gly157Glu＋Ser158Glu＋Asn198AspGly 61Asp＋Gln185Asp＋Asn198Gln＋Gln200Asp＋Thr207Asp＋Leu211SerAsp 58Glu＋Asn198Gln＋Gln200Glu＋Thr207Glu＋Leu211Ala＋Asn212GluGly 59Asp＋Ser 97Glu＋Pro129Asn＋Gly152Pro＋Ser182Glu＋Thr214AspGly 59Glu＋Thr 64Glu＋Pro127Gln＋Pro129Asn＋Ala131Gly＋Asn153GluPro127Glu＋Ser130Asp＋Ser154Glu＋Ile159Cys＋Ser210Glu＋Leu211SerAsp 58Glu＋Val 93Asn＋Ser 97Asp＋Gly100Glu＋Gly152Pro＋Asn198GluGly125Glu＋Gly155Asp＋Gly157Asn＋Gln200Glu＋Gly205Asn＋Ser210AspAla 96Ser＋Ser103Glu＋Leu124Asp＋Gln200Glu＋Ser210Glu＋Leu211CysSer101Asp＋Pro129Asp＋Gly155Asn＋Ala181Glu＋Ser182Asp＋Gly205ProAsp 58Glu＋Gly 61Glu＋Ala 96His＋Asn153Asp＋Val197Gln＋Gln200GluAsn 60Asp＋Ser 99Glu＋Gly155Pro＋Ala181Glu＋Val197Asn＋Asn212AspAsn 60Gln＋Gly 98Asn＋Ser130Asp＋Ala181Asp＋Ala209Asp＋Asn212AspGly 61Asp＋Ala131Asp＋Val197Ala＋Asn198Asp＋Gln200Asp＋Thr214AsnThr 64Asp＋Ser 99Asp＋Ser101Glu＋Ala181Gln＋Gln200Ser＋Ser210GluLeu124Ser＋Tyr161Glu＋Ser182Asp＋Val197Glu＋Thr207Gln＋Leu211AsnGly 95Glu＋Gly 98Asp＋Ser103Asp＋Ala181Gln＋Thr207Gln＋Asn212AspAsn 60Glu＋Ser 97Glu＋Ser101Glu＋Leu124Ala＋Pro127Glu＋Val197GlnVal 93Asp＋Ala 96Glu＋Ala131Asp＋Gly152Gln＋Gly196Ser＋Val197HisVal 93Gly＋Ala131Asp＋Ile159Gly＋Ser182Glu＋Asn198Gln＋Gly213AspSer130Asp＋Ser182Asp＋Gln185Ser＋Val197Asn＋Asn198Glu＋Ser210AspIle159Thr＋Ser182Asp＋Gln200Asp＋Gly205Asp＋Thr207Asn＋Asn212GluVal 93Thr＋Ser104Glu＋Pro127Ser＋Ser128Asp＋Ser130Glu＋Asn153GluAsn 60Glu＋Leu 94Asp＋Gln200Ser＋Gly205Asp＋Ser210Asp＋Leu211GlnGly 95Asp＋Ser101Asp＋Ser130Glu＋Asn198Asp＋Gln200Ser＋Leu211AlaSer104Asp＋Ser154Glu＋Thr207Asn＋Ala209Asp＋Ser210Glu＋Leu211AsnAsp 58Glu＋Gly 59Glu＋Ile105Leu＋Ser126Asp＋Phe183Asn＋Ser210AspGly 59Glu＋Asn 60Asp＋Gly 95Asn＋Pro129Glu＋Gly152Ser＋Gln185GluGly 59Asp＋Ser160Glu＋Gly196Ser＋Gly205Ser＋Ser210Asp＋Leu211AspAsn 60Glu＋Ser 99Asp＋Gly157Glu＋Phe183Leu＋Val197Asn＋Thr207GlnSer 99Glu＋Asn198Asp＋Ser206Glu＋Thr207Asp＋Tyr208Asn＋Leu211HisGly 98Glu＋Ser 99Asp＋Ala131Pro＋Asn153Glu＋Tyr161Pro＋Tyr208GluSer 97Glu＋Gly 98Glu＋Ser182Asp＋Thr207Gln＋Ser210Asp＋Gly213SerSer 97Asp＋Gly 98Asp＋Ile105Val＋Pro129Gly＋Gln200Asp＋Gly205GluAsp 58Glu＋Ser103Asp＋Ser104Asp＋Phe183Gly＋Val197Pro＋Asn198AspAsn 60Ser＋Ser 99Glu＋Gly100Asp＋Pro129Gly＋Gly205Asp＋Ser210GluSer 99Glu＋Ser154Asp＋Tyr161Cys＋Val197Ser＋Gln200Asp＋Ser210AspSer101Glu＋Gly155Asp＋Gln185Ser＋Asn198Gln＋Gln200Asp＋Ser210GluGly 59Asp＋Tyr102Asn＋Ser158Asp＋Gln200Asp＋Tyr208Val＋Ser210GluGly 59Asp＋Ala156Gln＋Ile159Asp＋Phe183Gly＋Gln200Asp＋Ser210AspPro127Ser＋Ser160Asp＋Ser182Asp＋Val197Gln＋Gln200Glu＋Ser210AspGln 57Asn＋Val 93Asp＋Pro127Glu＋Asn198Gln＋Gln200Glu＋Ser210GluLeu 94Glu＋Pro129Gln＋Ser154Asp＋Val197His＋Gln200Asp＋Ser210GluAla 96Ser＋Gly 98Gln＋Ser126Asp＋Ser154Asp＋Val197Thr＋Thr207AspGln 57Ser＋Ser104Asp＋Gly125Asp＋Ser160Glu＋Gly205Pro＋Ser210GluLeu 94Gln＋Ser 99Asp＋Gly155Pro＋Val193Asn＋Gln200Asp＋Thr207AspGly 95Glu＋Gln200Glu＋Thr202Ser＋Thr207Asp＋Leu211Gln＋Thr214GlnThr 64Gln＋Ala 96Thr＋Ser104Asp＋Gln200Asp＋Thr207Asp＋Thr214ProThr 64Glu＋Tyr102Cys＋Ala156Ser＋Phe183Glu＋Pro195Gln＋Thr207GluGly 98Glu＋Val197His＋Thr207Gln＋Ala209Ser＋Ser210Asp＋Gly213AspGly 95Ser＋Gly157Asp＋Phe183Glu＋Thr207Asp＋Ala209Gln＋Ser210AspAsp 58Glu＋Val 93Asp＋Ser128Glu＋Ala131Glu＋Gly196Gln＋Pro204AsnGln 57Ser＋Pro127Glu＋Gln185Asn＋Asn198Asp＋Gln200Glu＋Ser206GluVal 93Glu＋Gly152Asn＋Thr207Glu＋Ser210Asp＋Leu211Ser＋Gly213GluThr 64Gly＋Ser 99Glu＋Ser103Glu＋Pro127Asn＋Gly152Glu＋Ser158GluGly157Asn＋Ser158Glu＋Asn198Asp＋Ser206Asp＋Thr207Gln＋Ser210AspAla 96Asp＋Ser126Glu＋Ser160Glu＋Tyr161Leu＋Ser210Glu＋Gly213SerAsp 58Glu＋Ala 96Ser＋Ser101Asp＋Ser104Glu＋Ala209Gln＋Ser210AspGly 61Asp＋Thr 64Gln＋Val 93Cys＋Ile105Ser＋Leu211Val＋Asn212GluGly 61Glu＋Phe183Ser＋Tyr203Thr＋Thr207Asn＋Tyr208Gly＋Asn212GluSer 97Glu＋Ile105Cys＋Pro127Ser＋Ser160Asp＋Asn198Glu＋Ser210AspLeu 94Asp＋Ser130Glu＋Asn198Asp＋Pro204Asn＋Thr207Gln＋Ser210AspIle105Asp＋Leu124Pro＋Ile159Leu＋Asn198Glu＋Ser206Asp＋Leu211GluTyr102Gly＋Ala131Ser＋Ser182Glu＋Phe183Pro＋Thr207Glu＋Ser210AspAsn 60Gln＋Gly 61Glu＋Ser 97Asp＋Gly125Glu＋Tyr208Pro＋Ser210AspGly 98Gln＋Ser160Glu＋Ala181Thr＋Asn198Asp＋Ser206Asp＋Asn212GluLeu124Ala＋Ser128Glu＋Asn153Gln＋Gly205Asp＋Thr207Glu＋Asn212GluGly 59Glu＋Gly100Pro＋Ser103Glu＋Ile105Glu＋Val197Gly＋Ser210GluAsn 60Glu＋Ser 97Glu＋Pro127Asp＋Val193Met＋Thr207Asp＋Ala209AsnThr 64Gln＋Ala 96Asn＋Ser101Glu＋Ser130Asp＋Asn198Ser＋Leu211AspGly 61Pro＋Leu 94Glu＋Ser103Asp＋Ile159Pro＋Leu211Asn＋Asn212GluVal 93Cys＋Leu 94Asp＋Gly100Ser＋Ser103Glu＋Ser206Asp＋Tyr208GlySer130Asp＋Gly152Glu＋Ser154Glu＋Gly157Ser＋Ala181Pro＋Asn198GluGly 59Glu＋Ser130Asp＋Ser154Asp＋Gln185Asp＋Thr207Gly＋Leu211ProGln 57Glu＋Leu 94Ala＋Ser101Asp＋Ser126Glu＋Ala131Gln＋Ser154AspSer103Asp＋Ile105Met＋Gly125Asn＋Ser160Asp＋Val197Ser＋Thr214AspAsp 58Glu＋Val 93Asp＋Gly125Ser＋Gly152Ser＋Val193His＋Gln200GluAsp 58Glu＋Gly 98Asp＋Leu124Ser＋Gly125Glu＋Tyr161Met＋Gln200GluGly 61Asp＋Leu 94Ile＋Asn153Asp＋Ser158Asp＋Val197Ala＋Ser210GluAsn 60Ser＋Val 93Ser＋Ser103Glu＋Ser160Glu＋Asn198Asp＋Gln200AspAsn 60Gln＋Ser 97Asp＋Ser128Asp＋Asn198Glu＋Gln200Asp＋Ala209GlyVal 93Ser＋Tyr102Asn＋Ser104Asp＋Gly152Asp＋Asn198Glu＋Gln200GluGly 59Asp＋Ile105Met＋Ser158Glu＋Ile159Ser＋Asn198Glu＋Gln200AspGly 59Glu＋Ser101Glu＋Ile159Cys＋Asn198Glu＋Val199Gln＋Gln200AspGly 61Asp＋Pro127Asp＋Gln185Asp＋Asn198Ser＋Gln200Ser＋Ser210AspGly100Glu＋Ser104Glu＋Ser182Glu＋Thr207Gly＋Tyr208Met＋Ser210GluThr 64Gly＋Gly157Asp＋Ser182Glu＋Val197Gln＋Asn198Ser＋Ser210AspSer128Glu＋Ser130Asp＋Tyr161Val＋Asn198Glu＋Ser206Glu＋Gly213AsnGly 59Gln＋Thr 64Asp＋Ser 99Glu＋Asn198Asp＋Pro204Glu＋Thr207AsnGly 95Glu＋Ser126Asp＋Gln185Ser＋Val197Cys＋Gln200Glu＋Leu211GluGly 98Pro＋Ser 99Glu＋Ser158Asp＋Gln200Asp＋Ala209Ser＋Leu211AspSer101Glu＋Pro129Glu＋Asn153Ser＋Ser160Asp＋Thr207Glu＋Leu211CysAsn 60Glu＋Gly 98Asp＋Pro127Asp＋Ala131Gln＋Ala181Asp＋Asn198SerAsp 58Glu＋Ser126Asp＋Gly157Asn＋Thr207Ser＋Leu211Asp＋Gly213AspSer104Asp＋Leu124Gly＋Asn153Asp＋Ser182Glu＋Asn198Glu＋Thr207AsnGly 955er＋Ser103Glu＋Pro129Glu＋Ser160Asp＋Gln200Asp＋Leu211ProGly 59Glu＋Gly125Asp＋Pro127Glu＋Ala131His＋Ala209Asp＋Leu211IlePro127Glu＋Gly157Asn＋Pro204Gly＋Thr207Asp＋Ala209Asp＋Thr214AspGln 57Glu＋Leu 94Glu＋Ser103Asp＋Gly152Asp＋Gly205Ser＋Leu211AlaAla 96Glu＋Ser103Asp＋Gly152Gln＋Ser158Glu＋Ser160Asp＋Thr214AsnGly 61Glu＋Tyr102His＋Asn153Asp＋Phe183His＋Gln185Asn＋Ser210AspSer 97Asp＋Ser126Asp＋Pro129Glu＋Val197His＋Asn198Asp＋Gln200AsnAsn 60Glu＋Ala 96Pro＋Ile105Asp＋Leu124Ala＋Ser130Glu＋Ile159SerGln 57Glu＋Pro127Gly＋Ser128Glu＋Ala131Ser＋Ser210Asp＋Asn212GluAla 96Glu＋Tyr102Asp＋Ser128Glu＋Ala156Pro＋Tyr161Val＋Leu211ThrGln 57Ser＋Asp 58Glu＋Ser 99Asp＋Gln200Glu＋Tyr208Ser＋Asn212GluGly 59Asp＋Leu 94Met＋Ile105Ala＋Gly152Glu＋Ser182Glu＋Gln185SerAsn 60Glu＋Ala156Gly＋Val197Asp＋Asn198Ser＋Gln200Glu＋Ser206AspGly100Ser＋Ser101Asp＋Gly125Gln＋Ser126Asp＋Pro127Asn＋Asn198GluAla 96Glu＋Ser 99Asp＋Ser160Asp＋Ala181Gly＋Gly205Ser＋Ser210AspLeu124Asp＋Gly125Pro＋Ser130Glu＋Ser154Asp＋Gly205Gln＋Ala209ThrSer101Glu＋Gly152Glu＋Gly155Glu＋Asn198Gln＋Gln200Asp＋Asn212SerGly 95Asn＋Ser103Asp＋Gly152Glu＋Gly155Asp＋Tyr208Val＋Ser210GluSer 99Glu＋Gly100Ser＋Gly125Asn＋Gly152Asp＋Gly155Asp＋Gly205AspAla 96Glu＋Ser160Asp＋Tyr161Met＋Phe183Leu＋Gln185Asp＋Gly205AspLeu 94Glu＋Ser104Asp＋Gly157Glu＋Asn198Gln＋Thr207Asn＋Ala209AspThr 64Gln＋Ser101Glu＋Ile105Pro＋Gly125Ser＋Pro129Glu＋Thr214AsnGly 98Gln＋Tyr102Gln＋Gly125Asn＋Ser126Glu＋Ser158Glu＋Asn198GluLeu 94Asp＋Tyr161Ile＋Ala181Gly＋Phe183Asp＋Gln200Glu＋Thr214Asp表38优选的枯草蛋白酶309突变体-单突变型-Thr207GluSer210GluSer210AspSer210GlVVa1197Glu-双突变型-Gln200Glu＋Ser210GluVal199Leu＋Ser210GluVal199Leu＋Ser210AspPro204Ala＋Ala209ThrThr207Glu＋Ser210GluTyr208Phe＋Leu211AsnAla194Glu＋Ser210GluGln200Glu＋Tyr211AsnGln206Glu＋Thr207Glu-三突变型-Gln200Pro＋Gly205Ala＋Ser210GluThr207Glu＋Ser210Glu＋leu211AsnVal199Ile＋Pro204Asn＋Thr207GluGln200Glu＋Ser210Glu＋Leu211AspGln200Glu＋Thr207Glu＋Leu211GluGln200Glu＋Thr207Gln＋Ser210Glu-四突变型-Pro204Ala＋Thr207Glu＋Ser210Glu＋Leu211AsnGln206Glu＋Thr207Glu＋Ser210Glu＋Leu211Gly-五突变型-Val197Leu＋Pro204Ala＋Thr207Glu＋Ser210Glu＋Leu217AspAsn198Glu＋Gln200Glu＋Thr207Glu＋Ser210Glu＋Leu211Asp
II.洗涤组合物本发明的又一个方案是在用于清洁许多需要去清除蛋白质污渍的表面的洗涤组合物中，掺入一定有效量的一种或多种本发明的酶的突变体。此类清洁组成物包括清洁硬表面的洗涤剂组合物，形式不限(例如液体和颗粒状)；洗涤织物的洗涤剂，形式不限(例如颗粒状、液体和条状)；洗碟剂(形式不限)；口腔清洁剂，形式不限(例如牙粉、牙膏、漱口剂)假牙清洁剂，形式不限(例如液体、片状)；以及接触镜片的清洁剂，形式不限(例如液体、片状)。此处“有效量的酶的突变体”指在特定的清洁组成物中达到必需的酶活性所需的酶的突变体的量。这一有效量很容易由本领域中的普通技术人员确定，并由多个因素决定，比如所用的特定的酶的突变体，洗涤方法，洗涤组合物的特定组成，以及需要的组合物为液体还是干态(如颗粒状、条状)等。优选地发明的洗涤组成物包括0.0001％-10％的一种或多种酶的突变体，更优选的包括0.001％-1％，更优选的包括0.01％-0.1％。几个不同的利用此发明的酶的突变体的洗涤组合物的实施例在以下将作进一步讨论。所有部分，此处所用的百分比和比例除特别指明外，均指重量。
在此处，“非织物洗涤组合物”包括硬表面清洗剂，洗碟剂、口腔清洗剂，假牙清洁剂和接触镜片清洗剂。
A.用于硬表面、餐具、织物的洗涤组合物此发明中的酶可用于任何需要多泡沫和去除难溶底物的洗涤剂组合物。因而此发明中的酶的突变体可以与常规成分一起用来制成硬表面清洁剂(Clearers)、洗碟剂、织物洗涤剂、诸如此类的完全配方此类组合物可以是液体、颗粒状、条状等。此类组合物可按配方制造包含多达30-60％的表面活化剂的“浓缩”洗涤剂。
此处的洗涤组合物可任选的和优选包含阴离子、非离子、两性离子等形式的表面活化剂。此类表面活化剂一般在组合物中为5-35％的水平。
此处用的非限定性的表面活性剂包括常规的C11-G18烷基苯磺酸及伯位和随机烷基硫酸酯；C10-C18仲位(2，3)烷基硫酸盐其通式为CH3(CH2)x(CHOSO3)-M+CH3和CH3(CH2)y(CHOSO3-M+)CH2CH3，其中x和(y＋1)为至少约为7的整数。优选不小于9，而M为水溶性阳离子，尤其是钠离子；C10-C18烷基烷氧基硫酸盐(尤其是EO1-5乙氧基硫酸盐)。C10-C18烷基烷氧基羧酸盐(尤其是EO1-5乙氧基羧酸盐)，C10-C18烷基多聚葡萄糖苷及它们相应的硫酸聚糖苷；C12-C18α-硫酸脂肪酸酯，C12-C18烷基苯烷氧基化物(尤其是乙氧基化物和混合的乙氧基/丙氧基)C12-C18甜菜碱和磺基甜菜碱(“Sulfaines”)，C10-C18胺氧化物，等等。此处优选烷基烷氧基硫酸盐(AES)和烷基烷氧基羧酸盐(AEC)。(此类表面活化剂及以上所述胺氧化物和/或甜菜碱或磺基甜菜碱表面活化剂的使用，可根据配方需要优选地)其他常规有用的表面活化剂在标准的教科书中均有列出。包括C10-C18N-甲基葡萄糖酰胺(glucamide)的特殊用途的表面活化剂已公开于美国专利，5,194,639，Conner等，1993年3月16日出版。此外引入作为参考。
在洗涤剂洗涤组合物中有用的多种其他成分均可包含在这里所说的组合物中，包括其他活性成分，载体，水溶助长剂hydrtropes、操作助剂、染料或色素，液体制剂的溶剂等等。如果需额外的泡沫增强，泡沫活化剂如C10-C16烷基醇酰胺可引入到组合物，通常在1％-10％左右的水平。C10-C14单乙醇酰胺和双乙醇酰胺代表了此类泡沫活化剂的典型类型。使用此类泡沫活化剂高泡沫粘附表面活化剂。如上述的胺氧化物，甜菜碱和磺基甜菜碱，亦是有利的。如果需要，可溶性的镁盐如MgCl2，MgSO4等，通常按0.1-2％左右水平加入来提供更多的泡沫。
此处的液体洗涤剂组合物可包含水和其他溶剂作为载体。低分子量伯醇或仲醇例如甲醇，乙醇，丙醇和异丙醇都是合适的。一元醇类是溶解表面活性剂的优选地，但多元醇如包括2-6个碳原子和2-6个羟基基团(例如1，3-丙乙醇，乙二醇，甘油，1，2-丙二醇)的亦可使用。组合物可包含5％-90％左右，通常的为10-50％的此类载体。
此处的洗涤剂组合物将优选地用于水环境中清洗操作而配制，洗涤用水pH在6.8到11.0左右，因而成品通常在此范围内配制。控制pH在建议范围内的技术包括运用缓冲液、碱、酸、等，这些是本领域技术人员所熟知的。
当配制此发明的硬表面和织物洗涤组合物，配制人也许希望使用重量比占5-50％左右的不同的材料，典型的材料包括1-10微米沸石，聚羧化物如枸橼酸盐和氧化二琥珀酸盐，层状硅酸盐、磷酸盐等，其他的常规材料在标准的配方中均有列出。
同样地，配制人可能希望在此类组合物中使用其它的各种酶，如纤维素酶、脂酶、淀粉酶和蛋白酶。通常重量范围在0.001％-1％左右。各种洗涤和用于织物的酶在洗衣洗涤剂领域都是众所周知的。
各种漂白化合物，如过碳酸盐。过硼酸盐等，可用于此类洗涤组合物，通常按重量计占1％-15％。如果需要，此类组合物亦可包含漂白活化剂如四乙酰基乙二胺。硫酸壬酰羟苯酯等，这也是本领域中所周知的。使用按重量计占通常在1％-10％左右。
各种污物去除剂，尤其是阴离子寡酯类；各种螯合剂。尤其是氨基磷酸盐和乙二胺二琥珀酸盐；各种粘性污物去除剂。尤其是乙氧基四亚乙基戊胺；各种分散剂，尤其聚丙烯酸酯和聚Asp；各种增白剂，尤其是阴离子增白剂；各种消泡剂，尤其是硅氧烷和仲醇；各种织物软化剂，尤其是绿土等均可用于此类组成物，重量范围在1％-35％左右。标准的配方和公开的专利包含多种，详细此类常规原料的叙述。
酶稳定剂亦可用于此发明的洗涤组合物中，此类酶稳定剂包括丙二醇(优选1％-10％)，甲酸钠(优选0.1％-1％)，和甲酸钙(优选0.1％-1％)。
1.硬表面洗涤组合物此处“硬表面洗涤组合物”指液体如颗粒状洗涤组合物，用于清洗地板、墙壁、浴池贴砖等硬表面。此发明的硬表面洗涤组合物包含一定有效量的一种或多种此发明中的酶突变体，活性酶重量的优选范围约为0.001％-10％，更优选范围约为0.01％-5％，最优选约为0.05％-1％。除了包含一种或多种此发明中的酶突变体，此类硬表面洗涤组合物通常包括表面活性剂和水溶性螯合剂材料。然而在某些特殊产品如窗户喷雾清洁剂中，表面活性剂有时不被使用，因为它们可能在玻璃表面产生膜状/条纹状残痕。
当存在时，表面活性剂成分在此处组合物中可占少至0.1％，但通常组合物将占约0.25％-10％，更优选范围约为1％-5％。
通常情况组合物将包含约0.5％-50％的洗涤助洗剂，优选范围1％-10％。优选pH8-12。常规的pH调节剂如氢氧化钠，碳酸钠或盐酸亦可在需要时使用。
溶剂可包含在组合物中，有用的溶剂包括但不局限于二乙二醇醚如乙二醇单己基醚，乙二醇单丁基醚，丙二醇单丁基醚，乙二醇单己基醚、丙二醇单丁基醚，二丙二醇单丁基醚、和二醇如2，2，4-三甲基-1，3-戊二醇和2-乙基-1，3-己二醇。使用时，此类溶剂通常标准在0.5-15％左右，优选约3-11％。
另外，高挥发性溶剂如异丙醇或乙醇亦可用此组合物中，以便当组合物“完全力量”(“full strength”)施用于表面外，表面不被清洗时，促进组合物在表面快速蒸发。使用时，挥出性溶剂通常在大约2％-12％。
此发明的硬表面洗涤组合物实施方案在下面这些例子中阐明。
实施例7-12液体硬表面清洁组合物实施例号组分7 8 9 101112Thr207G1u 0.050.50 0.02 0.03 0.10 0.03Val199Leu＋Ser210Asp- - - - 0.20 0.02Na2DIDA*EDTA**- - 2.90 2.90 - -枸椽酸钠- - - - 2.90 2.90C12烷基苯磺酸钠1.95- 1.95 - 1.95 -C12烷基硫酸钠 - 2.20 - 2.20 - 2.20C12(乙氧基)磺酸钠***- 2.20 - 2.20 - 2.20二甲胺氧化物- 0.50 - 0.50 - 0.50异丙苯磺酸钠1.30- 1.30 - 1.30 -己基卡必醇***6.306.30 6.30 6.30 6.30 6.30水****平衡至100％*N-二乙二醇-N，N-亚氧基二乙酸二钠**乙二胺二乙酸四钠***二乙二醇-单己基-醚****所有配方调pH至7在实施例7-10，表3-38列举的枯草蛋白酶309突变体被突变体Thy-207Glu所代替，结果基本相似。
在实施例11-12，表3-38中所列举的任何枯草蛋白酶309联合突变体被突变体Val199Leu＋Ser210Asp所代替，结果基本相似。
实施例13-18洗涤硬表面及去除家居霉的喷雾组合物实施例号组分 13 14 15 16 17 18Gln200Glu＋Thr207Glu 0.50 0.05 0.60 0.30 0.20 0.30Ser210G1u - - - - 0.30 0.10辛烷基硫酸钠 2.00 2.00 2.00 2.00 2.00 2.00十二烷基硫酸钠 4.00 4.00 4.00 4.00 4.00 4.00氢氧化钠 0.80 0.80 0.80 0.80 0.80 0.80硅酸钠 0.04 0.04 0.04 0.04 0.04 0.04香料 0.35 0.35 0.35 0.35 0.35 0.35水平衡至100％在实施例13-16中，表3-38所列举的枯草蛋白酶309突变体被Glu200GLu＋Thr207Glu代替，结果基本相似。
在实施例17-18中，表3-38所列举的任何枯草蛋白酶309复合突变体，被突变体Gln200GLu＋Thr207Glu和Ser210Glu代替，结果基本相似。
2.餐具洗涤组合物本发明的另一个方案中，餐具洗涤组合物包含此发明的一种或多种酶的突变体。此处“餐具洗涤组合物”指所有形式清洁餐具的组合物，包括但不限于颗粒和液体形成。此发明的餐具洗涤组合物方案在下面实施例中阐明。
实施例19-24餐具清洗组合物实施例号组分 19 20 21 22 23 24Gln200Pro＋Gly205Ala 0.05 0.50 0.02 0.40 0.10 0.03＋Ser201AspVal199Leu＋Ser210Asp - - - - 0.40 0.02C12-C14N-甲基葡萄糖 0.90 0.90 0.90 0.90 0.90 0.90酰胺C12乙氧基(1)硫酸盐 12.00 12.00 12.0012.0012.00 12.002-甲基十一烷酸4.504.504.504.50 4.504.50C12乙氧基(2)羧酸盐 4.504.504.504.50 4.504.50C12醇乙氧基化物(4) 3.003.003.003.00 3.003.00C12胺氧化物 3.003.003.003.00 3.003.00异丙苯基磺酸钠2.002.002.002.00 2.002.00乙醇 4.004.004.004.00 4.004.00Mg++(如MgCl2) 0.200.200.200.20 0.200.20Ma++(如CaCl2) 0.400.400.400.40 0.400.40水平衡至100％产品pH调为7在实施例19-22中，表3-38所列举的枯草蛋白酶309突变体被突变体Glu200pro＋Gly205Ala＋Ser210Asp所代替，结果基本相似。
在实施例23-24中，表3-38所列举的任何复合突变体被突变体Glu200pro＋Gly205Ala＋Ser210Asp和Val199Leu＋Ser210Asp代替，结果基本相似。
3.织物组合物本发明的另一个方案中，织物洗涤组合物包括此发明的一个或多个酶突变体，此处“织物洗涤组合物”指所有形式用于清洁织物的去污组合物，包括但不限于颗粒状，液体和条状，优选的洗涤组合物为液体形式。
a.颗粒状织物洗涤组合物此发明的颗粒状织物洗涤组合物包含一定有效量一种或多种酶突变体，优选活性酶重量范围大约为0.001％-10％，更优选0.005％-5％，最优选大约0.01％-1％。除了一种或多种酶突变体，颗粒状织物洗涤组合物通常包括至少一种表面活性剂，一种或多种洗涤助洗剂，以及某些情况还包括一种漂白剂。
此发明的颗粒状织物洗涤组合物方案在下列实施例中阐明。
实施例25-28颗粒状织物洗涤组合物实施例号组分25 26 27 28Ser99Asp0.100.20 0.03 0.05Ser99Gly- -0.02 0.05C13线性烷基苯磺酸盐22.00 22.0022.0022.00磷酸盐(如三聚磷酸钠)23.00 23.0023.0023.00碳酸钠 23.00 23.0023.0023.00硅酸钠 14.00 14.0014.0014.00沸石8.208.20 8.20 8.20螯合剂(二亚乙基 0.400.40 0.40 0.40三胺五乙酸)硫酸钠 5.505.50 5.50 5.50水 平衡至100％在实施例25-26中，表3-38所列举的枯草蛋白酶309突变体被突变体Ser99Asp所代替，结果基本相似。
在实施例27-28中，任何表3-38所列举的枯草蛋白酶309突变体的复合突变体被突变体Ser99Asp和Ser99Gly代替，结果基本相似。
在实施例29-30中，表3-38所列举的枯草蛋白酶309突变体被突变体Glu200Glu＋Thy207Glu＋Ser210Glu所代替，结果基本相似。
实施例29-32颗粒状织物洗涤组合物实施例号组分 29 3031 32Gln200Glu＋Thr207Glu＋0.100.20 0.030.05Ser210GluAsn74Asp＋Pro204Ala＋ --0.020.05Thr207GluC12烷基苯磺酸盐 12.0012.0012.00 12.00沸石A(1-10μm) 26.00 26.00 26.00 26.002-丁基辛酸 4.004.004.004.00C12-C14(2，3)-烷基硫酸钠 5.005.005.005.00盐枸椽酸钠5.005.005.005.00荧光增白剂 0.100.100.100.10硫酸钠 17.00 17.00 17.00 17.00水和其它次要成分 平衡至100％在实施例31-32中，任何表3-38所列举的枯草蛋白酶309复合突变体被突变体Glu200Gou＋Thr207Glu＋Ser210Glu和Asn74Asp＋Pro204Ala＋Thr207Glu，结果基本相似。
实施例33-36颗粒状织物洗涤组合物实施例号组分 34 35 36 37Leu94Gly＋Gln200Glu0.100.20 0.030.05Gln57Ser＋Leu94Gly＋ - -0.020.05Gln200GluC13线性烷基苯磺酸酸盐 22.00 22.0022.00 22.00磷酸盐(如三聚磷酸钠) 23.00 23.0023.00 23.00碳酸钠 23.00 23.0023.00 23.00硅酸钠 14.00 14.0014.00 14.00沸石 8.208.20 8.208.20螯合剂(二亚乙基三胺五乙0.400.40 0.400.40酸)硫酸钠 5.505.50 5.505.50水 平衡至100％在实施例35-36中，表3-38列举的任何枯草蛋白酶复合突变体被Leu94Gly＋Gln200Glu和Gln57Ser＋Leu94Gly＋Gln200Glu代替，结果基本相似。
实施例37-40颗粒状织物洗涤组合物实施例号组分37 38 39 40Asn74His＋Gln57Ser＋Asn60Ser 0.10 0.200.030.05＋Leu94Gly＋GlnGluVal93Gln＋Tyr102Cys＋Ser154Glu-- 0.020.05＋Asn198Gln＋Thr207GlyC12烷基苯磺酸盐 12.0012.00 12.00 12.00沸石A(1-10um) 26.0026.00 26.00 26.002-丁基辛酸4.00 4.004.004.00C12-C14仲(2，3)-烷基磺酸钠盐5.00 5.005.005.00枸椽酸钠 5.00 5.005.005.00荧光增白剂0.10 0.100.100.10硫酸钠17.0017.00 17.00 17.00水和其它次要成分 平衡至100％在实施例37-38中，表37-38列举的枯草蛋白酶309突变体被Asn74His＋Gln57Ser＋Asn60Ser＋Leu94Gly＋Gln200Glu代替，结果基本相似。
在实施例39-40中，表3-38所列举的任何枯草蛋白酶309复合突变体被突变体Asn74His＋Gln57Ser＋Asn60Ser-Len94Gly＋Gln200Glu和Val93Glu＋Tyr102lys＋Ser154Glu＋Asn198Gln＋Thr207Gly代替，结果基本相似。
实施例41-42颗粒状织物洗涤组合物实施例号组分41 42线性烷基苯磺酸盐11.410.70牛脂烷基硫酸盐 1.802.40C14-C15烷基硫酸盐 3.003.107倍乙氧基化C14-C15醇 4.004.00牛脂11倍乙氧基化醇 1.801.80分散剂 0.070.1硅氧烷 0.800.80枸椽酸三钠 14.00 15.00枸椽酸 3.002.50沸石32.532.50顺丁烯二酸丙烯酸共聚物 5.005.00二亚乙基三胺五亚甲基膦酸1.000.20Ser210Asp 0.300.30脂酶0.360.40淀粉酶 0.300.30硅酸钠 2.002.50硫酸钠 3.505.20聚乙烯吡咯烷酮 0.300.50过硼酸盐0.5 1苯酚磺酸0.1 0.2过氧化物酶 0.1 0.1其它次要成分至100 至100实施例43-44颗粒状织物洗涤组合物实施例号组分 43 44C12线性烷基苯磺酸钠 6.5 8.0硫酸钠 15.018.0沸石A 26.022.0次氮基三乙酸钠 5.0 5.0聚乙烯吡咯烷酮 0.5 0.7四乙酰基乙二胺 3.0 3.0硼酸 4.0 -过硼酸盐 0.5 1苯酚磺酸盐 0.1 0.2Val199Leu＋Ser210Glu 0.4 0.4填充剂(如，硅酸盐、碳酸盐、至100至100香料、水)实施例45浓缩颗粒状织物洗涤组合物组分 重量百分比烷基硫酸盐8.0烷基乙氧基硫酸盐 2.03和7倍乙氧基化的C25和C45醇的混合物 6.0多羟基脂肪酸酰胺 2.5沸石 17.0层状硅酸盐/枸椽酸盐 16.0碳酸盐7.0顺丁烯二酸丙烯酸共聚物5.0污物去除聚合物0.4羧甲基纤维素 0.4聚(4-乙烯吡咯烷酮)-N氧化物0.1乙烯咪唑和乙烯吡咯烷酮共聚物 0.1PEG2000.2Asn74Asp＋Val197Glu＋Gln200Glu＋Ser210Glu 0.5脂酶 0.2纤维素酶 0.2四乙酰基乙二胺6.0过碳酸盐 22.0乙二胺二丁二酸0.3消泡剂3.54，4’-双(2-吗啉代-4-苯胺基均三嗪-6-基氨 0.25基)芪-2，2’-二磺酸二钠4，4’-双(2-硫化肉桂基(styril))联苯二钠0.5水、香料和其它次要成分 至100实施例46颗粒状织物洗涤组合物组分 重量百分比线性烷基苯磺酸盐 7.6C16-C18烷基硫酸盐 1.37倍乙氧基C14-C15醇 4.0椰子烷基二甲基羟乙基氯化铵1.4分散剂0.07硅氧烷流体0.8枸椽酸三钠5.0沸4A 15.0顺丁烯二酸丙烯酸共聚体4.0二亚甲基三胺五亚甲基膦酸 0.4过硼酸盐 15.0四乙酰基乙二胺5.0蒙脱石绿土10.0聚(氧乙烯)(分子量300,000) 0.3Tyr208Phe＋Leu211Asn 0.4脂酶 0.2淀粉酶0.3纤维素酶 0.2硅酸钠3.0硫酸钠10.0羧甲基纤维素 0.2增白剂0.2水、香料和其它次要成分至100
实施例47颗粒状织物洗涤组合物组分 重量百分比线性烷基苯磺酸盐6.92牛脂烷基硫酸盐 2.057倍乙氧基化C14-15醇 4.43倍乙氧基化C12-C15烷基乙氧基化硫酸酸 0.16沸石20.2枸椽酸盐5.5碳酸盐 15.4硅酸盐 3.0顺丁烯二酸丙烯酸共聚物 4.0羧甲基纤维素0.31污物去除聚合物 0.30Asn74His＋Val197Glu＋Pro204Ala 0.2＋Ala209Thr＋Ser210Glu脂酶0.36纤维素酶0.13四水合过硼酸盐 11.64单水合硼酸盐8.7四乙酰基乙二胺 5.0二亚乙基三胺五甲基磷酸 0.38硫酸镁 0.40增白剂 0.19香料、硅氧烷、消泡剂0.85其它次要成分至100b.液体织物洗涤组合物此发明的液体织物洗涤组合物包含一定有效量的此发明中的一种或多种酶突变体，组合物中活性酶优选重量范围大约为0.005％-5％，更优选大约为0.01％-1％。此种液体织物洗涤组合物通常另外包括一种阴离子表面活性剂，脂肪酸，一种水溶性洗涤助洗剂和水。此发明的液体织物洗涤组合物方案在下列实施例中阐明。
实施例48-52液体织物洗涤组合物实施例号组分 48 49 50 51 52Pro204Ala＋Ala209Thr 0.5 0.030.300.03 0.10Gln200Glu＋Thr207Glu - - - 0.01 0.20＋Ser210GluC12-C14烷基硫酸钠 20.00 20.00 20.00 20.00 20.002-丁基辛酸 5.005.005.005.00 5.00枸椽酸钠 1.001.001.001.00 1.00C10醇乙氧基化物(3) 13.00 13.00 13.00 13.00 13.00单乙醇胺(Monetharolamine)2.502.502.502.50 2.50水/丙二醇/乙醇(100∶1∶1) 平衡至100％在实施例48-50中，表3-38列举的枯草蛋白酶309突变体被Pro204Ala＋Ala209Thr代替，结果基本相似。
在实施例51-52中，表3-38列举的任何枯草蛋白酶309复合突变体被Pro204Ala＋Ala209Thr和Glu200Glu＋Thr207Glu＋Ser210Glu代替，结果基本相似。
实施例53-57液体织物洗涤组合物实施例号组分 53 54 55 56 57Thr102Cys＋Ile105Val 0.5 0.030.300.030.10＋Leu124Ile＋Ser154Glu＋Asn198Gln＋Thr207GlyAsn74Asp＋Ser97Asp- - - 0.010.20＋Gln57Ser＋Asn60SerC12-C14烷基硫酸钠 20.00 20.00 20.00 20.00 20.002-丁基辛酸5.005.005.005.005.00枸椽酸钠 1.001.001.001.00 1.00C10醇乙氧基化物(3) 13.00 13.00 13.00 13.00 13.00单乙醇胺 2.502.502.502.50 2.50水/丙二醇/乙醇(100∶1∶1) 平衡至100％在实施例53-55中，表3-38列举的枯草蛋白酶309突变体被Tyr102Cys＋Ile105Val＋Leu124Ile＋Ser154Glu＋Asn198Gln＋Thr207Gly代替，结果基本相似。
在实施例56-57中，表3-38列举的任何枯草蛋白酶309复合突变体被Tyr102Cys＋Ile105Val＋Leu124Ile＋Ser154Glu＋Asn198Gln＋Thr207Gly和Asn74Asp＋Ser97Asp＋Gln57Ser＋Asn60Ser代替，结果基本类似。
实施例58-59颗粒状织物洗涤组合物实施例号组分 58 59C12-C14链烯基琥珀酸 3.0 8.0单水合枸椽酸 10.015.0C12-C15烷基硫酸钠8.0 8.02倍乙氧基化C12-C15醇的硫酸钠 - 3.02倍乙氧基化C12-C15醇 - 8.05倍乙氧基化C12-C15醇 8.0 -二亚乙基三胺五(亚甲基磷酸) 0.2 -油酸 1.8 -乙醇 4.0 4.0丙二醇 2.0 2.0Asn74Asp＋Ser210Glu0.2 0.2聚乙烯吡咯烷酮 1.0 2.0消泡剂 0.150.15NaOH 至pH7.5过硼酸盐 0.5 1苯酚磺酸盐 0.1 0.2过氧化物酶 0.4 0.1水和其它次要成分 至100份此处58和59实施例中，表3-38列举的枯草蛋白酶309突变体被Asn74Asp＋Ser210Glu代替，结果基本相似。
实施例60-62液体织物洗涤组合物实施例号组分 60 61 62枸椽酸 7.103.003.00脂肪酸 2.00- 2.00乙醇 1.933.203.20硼酸 2.223.503.50单乙醇胺 0.711.091.091，2丙二醇 7.898.008.00乙丙苯基硫酸钠 1.803.003.00甲酸钠 0.080.080.08氢氧化钠 6.703.803.80硅抗泡沫剂 1.161.181.18Ser210Glu0.0145 - -Asn74Asp＋Ser97Asp＋Ser210Glu- 0.0145 -Gln200Glu＋Ser210Glu - - 0.0145脂酶 0.200 0.200 0.200纤维素酶 - 7.507.50污物去除聚合物 0.290.150.15抗泡沫剂 0.060.085 0.085增白剂36 0.095 - -增白剂3 - 0.050.05C12烷基苯磺酸 9.86- -C12-C15烷基聚乙氧基(2.5) 13.80 18.00 18.00硫酸盐C12葡萄糖酰胺 - 5.005.00C12-C13聚乙氧基化物2.00 2.00 2.00水、香料和其它次要成分 平衡至100％c.条状(bar)织物洗涤组合物此发明中适于手洗污损织物的条状织物洗涤组合物包含一定有效量的此发明的一种或多种酶突变，组合物中优选按重量计占0.001％-10％，更为优选约为0.01-1％。
此发明的条块织物洗涤组合物方案在下列实施例中阐明。
实施例63-66条状(bar)洗涤组合物实施例号组分 63 64 65 66Val197Glu 0.3- 0.10.02Tyr208Phe＋Leu211Asn - - 0.40.03C12-C16烷基硫酸钠 20.0 20.020.0 20.00C12-C14-甲基葡萄糖酰胺 5.05.0 5.05.00C11-C13烷基苯磺酸钠 10.0 10.010.0 10.00碳酸钠25.0 25.025.0 25.00焦磷酸钠 7.07.0 7.07.00三磷酸钠 7.07.0 7.07.00沸石A(0.1-10u)5.05.0 5.05.00羧甲基纤维素 0.20.2 0.20.20聚丙烯酸盐(分子量1400)0.20.2 0.20.20椰子单乙醇胺 5.05.0 5.05.00增白剂、香料 0.20.2 0.20.20碳酸钙1.01.0 1.01.00硫酸镁1.01.0 1.01.00水4.04.0 4.04.00填料*平衡至100％*表示可从方便的材料如CaCO3、滑石、粘土、硅酸盐等中选取在实施例63-64中，表3-38列举的枯草蛋白酶309突变体被Val197Glu代替，结果基本相似。
在实施例65-66中，表3-38列举的任何枯草蛋白酶309复合突变体被Val197Glu和Tyr208Phe＋Leu211Asn代替，结果基本相似。
实施例67-70条状(bar)洗涤组合物实施例号组分 67 68 69 70Asn 60Ser＋Val93Gln＋Gly213Asp 0.3- 0.10.02Val93Gln＋Tyr102Cys＋Thr207Gly - 0.30.40.03＋Gly213AspC12-C16烷基硫酸钠 20.0 20.0 20.0 20.00C12-C14N-甲基葡萄糖酰胺5.05.05.05.00C11-C13烷基苯硫酸钠10.0 10.0 10.0 10.00碳酸钠 25.0 25.0 25.0 25.00焦磷酸钠 7.07.07.07.00三磷酸钠 7.07.07.07.00沸石A(0.1-10u) 5.05.05.05.00羧甲基纤维素 0.20.20.20.20聚丙烯酸盐(分子量1400) 0.20.20.20.20椰子单乙醇酰胺 5.05.05.05.00增白剂，香料 0.20.20.20.20碳酸钙 1.01.01.01.00硫酸镁 1.01.01.01.00水 4.04.04.04.00填料*平衡至100％*表示可从方便的材料如CaCO3、滑石、粘土、硅酸盐等中选取在实施例67中，表3-38列举的枯草蛋白酶309的突变体被Asn60Ser＋Val93Gln＋Gly213Asp代替，结果基本相似。
在实施例68中，表3-38列举的枯草蛋白酶309的突变体被突变体Val93Gln＋Tyr102Cys＋Thr207Gly＋Gly213Asp代替，结果基本相似。
在实施例69-70中，表3-38列举的任何枯草蛋白酶309复合突变体被Asn60Ser＋Val93Gln＋Gly213Asp和Val93Gln＋Tyr102Cys＋Thr207Gly＋Gly213Asp代替，结果基本类似。
B.其他洗涤组合物除了上面讨论的清洁硬表面，洗涤餐具和清洗织物的组合物外，此发明中的一种或多种酶突变体亦可被引人到其他多种需要水解不溶性底物的洗涤组合物中。此类洗涤组合物包括但不限于口腔洗涤组合物，假牙洗涤组合物，和接触镜片洗涤组合物。
1.口腔清洁组合物在此发明的另一个方案中，本发明中的一定的药理学上可接受的量的一种或多种酶突变体被用于对牙齿和假牙上去除蛋白污物的组合物中。此处所用的“口腔洗涤组合物”指牙粉、牙膏、牙凝胶、牙粉、漱口剂、口腔喷雾剂、口腔凝胶、口香糖、锭剂、香粉、药片、生物凝胶、预防性膏、牙齿处理液等。口腔洗涤组合物优选按重量计包含约0.0001％-20％的一种或多种酶突变体，更优选为约0.001％-10％，最优选约0.01％-5％，和一种药理学上可接受的载体。此处“药理学上可接受”意思为此术语描述的药物、药剂、或惰性成分适于接触人类和较低等动物组织的使用，而没有过分的毒性、不适应性、不稳定性、刺痒、过敏反应等等，具有合理的利益/风险比例。
通常按重量计，对于口腔洗涤组合物中口腔清洁成分的清洁载体，药理学可接受的量一般为50-99.9％左右，优选为大约65％-99.99％，更为优选为大约65％-99％。
可包括在本发明口腔清洗组合物中的药理学上可接受的载体成分和可任选的组分，对于本领域的技术人员是众所周知的。在口腔清洗组合物中有用的诸多种组合物类型，载体成分和选择成分已公开于1992年3月17日公开的美国专利5,096,700，Seibel；1991年7月2日公开的美国专利5,028,414，Saurpathkumar；和1992年7月2日公开的美国专利5,028,415 Benedizt，Bush和Sunberg。所有这些在此引入作为参考。
此发明中的口腔洗涤组合物方案在下列实施例中阐明。
实施例71-74牙粉组合物实施例号组分 71 72 73 74Val199Leu＋Pro204Ala＋Thr207Glu 2.0003.5001.5002.000＋Ser210Glu山梨糖醇(70％水溶液) 35.000 35.000 35.000 35.000PEG-6*1.0001.0001.0001.000硅土牙磨料**20.000 20.000 20.000 20.000氟化钠0.2430.2430.2430.243二氧化钛 0.5000.5000.5000.500糖精钠0.2860.2860.2860.286烷基硫酸钠(27.9％水溶液) 4.0004.0004.0004.000香料 1.0401.0401.0401.040羧乙烯基聚合物***0.3000.3000.3000.300角叉菜胶(carrageenan)0****0.8000.8000.8000.800水 平衡至100％*分子量为600的聚乙二醇**J.M.Huber提供的鉴定为Zeodent119的沉淀的硅土****由B.F.Goodrich化学公司提供****少量角叉菜胶由Hercules化学公司提供在实施例71-74中，表3-38所列举的枯草蛋白酶309突变体被突变体Val199Leu＋Pro204Ala＋Thr207Glu＋Ser210Glu取代，结果基本相似。
实施例75-78漱口剂组合物实施例号组分 75 76 77 78Ser210Asp3.007.501.005.00SDA40酒精8.008.008.008.00香料0.080.080.080.08乳化剂 0.080.080.080.08氟化钠 0.050.050.050.05甘油10.00 10.00 10.00 10.00增甜剂 0.020.020.020.02苯甲酸 0.050.050.050.05氢氧化钠0.200.200.200.20染料0.040.040.040.04水 平衡至100％在实施例75-78中，表3-38所列举的枯草蛋白酶309突变体被突变体Ser210Asp取代，结果基本相似。
实施例79-82锭剂组合物实施例号组分 79 80 81 82Tyr208Phe＋Leu211Asn 0.01 0.030.10 0.02山梨糖醇 17.5017.50 17.50 17.50甘露醇 17.5017.50 17.50 17.50淀粉 13.6013.60 13.60 13.60增甜剂 1.20 1.201.20 1.20香料 11.7011.70 11.70 11.70色素 0.10 0.100.10 0.10玉米糖浆平衡至100％在实施例79-82中，表3-38所列举的枯草蛋白酶309突变体被突变体Tyr208Phe＋Leu211Asn代替，结果基本相似。
在实施例83-86中，表3-38所列举的枯草蛋白酶309突变体被突变体Vla199Net＋Pro204Ala＋Thr207Glu代替，结果基本相似。
实施例83-86口香糖组合物实施例号组分 83 84 85 86Val199Met＋Pro204Ala＋Thr207Glu0.030.020.10 0.05山梨糖醇晶体 38.44 38.40 38.40 38.40Paloja-T胶基*20.00 20.00 20.00 20.00山梨糖醇(70％水溶液) 22.00 22.00 22.00 22.00甘露醇 10.00 10.00 10.00 10.00甘油 7.567.567.56 7.56香料 1.001.001.00 1.00*L.A Dreyfus公司提供2.假牙洗涤组合物在此发明的另一个方案中，用来清洁口腔外的假牙的洗涤组合物包括本发明的一种或多种酶突变体。此类假牙清洗组合物包含一定有效量的一种或多种酶突变体，组合物和假牙清洁载体按重量计优选为大约0.0001％-50％的一种或多种酶突变体，更为优选约为0.001％-35％，更优选为大约0.01％-20％。不同假牙洗涤组合物形式如泡腾药片等在本领域内是周知的(参看美国专利5,055,305 Young中的例子，此处引入以为参考)，而且普遍适合于引入一种或多种酶突变体来清除假牙上的蛋白污物。
本发明的假牙洗涤组合物方案在下列实施例中阐明。
实施例87-90双层泡腾假牙清洁片剂实施例号组分 87 88 89 90酸层Ser210Glu1.0 1.5 0.010.05酒石酸 24.024.024.00 24.00碳酸钠 4.0 4.0 4.004.00磺酸 10.0 10.010.00 10.00聚乙二醇20,0004.04.0 4.004.00碳酸氢钠 24.5 24.524.50 24.50过硫酸钾 15.0 15.015.00 15.00焦硫酸钠 7.07.0 7.007.00焦化硅胶(Pyrogenic silica)2.02.0 2.002.00TEAD*7.07.0 7.007.00蓖麻酰硫代琥珀酸盐0.50.5 0.500.50香料 1.01.0 1.001.00碱层单水合过硼酸钠32.0 32.032.00 32.00碳酸氢钠 19.0 19.019.00 19.00EDTA 3.03.0 3.003.00三磷酸钠 12.0 12.012.00 12.00聚乙二醇20,0002.02.0 2.002.00过硫酸钾 26.0 26.026.00 26.00碳酸钠2.02.0 2.002.00焦化硅胶 2.02.0 2.002.00染料/香料 2.02.0 2.002.00*四乙酰基乙二胺在实施例87-90中，表3-38所列举的枯草蛋白酶309突变体被突变体Ser210Glu代替，结果基本相似。
3.接触镜片清洗组合物此发明的另一个方案中，接触镜片清洗组合物包括本发明的一种或多种酶突变体。此类接触镜片洗涤组合物包括一定有效量的一种或多种酶突变体，组合物和接触镜片清洁载体中按重量计，优选一种或多种酶突变体的量约为0.01％-50％，更优选约为0.01％-20％，最为优选约为1％-5％。不同的接触镜片清洗组合物形式如药片、液体等在本领域内为众所周知(例子可参看美国专利4,863,627，Davies，Meaken和Rees；1989年9月5日出版；美国专利研究32,672，Huth，Lam和Kirai，1988年5月22日再版；美国专利4,609,493，Schafer.1986年9月2日出版；美国专利4,690,793，Ogunbiyi，和Smith.1987年9月1日出版；美国专利4,614,549，Ogunbiyi，Riedhammer和Smith.1986年9月30日出版；以及美国专利4,285,738，Ogara，1981年8月21日公开；上述均在此处引入以为参考)，而且一般均适合于引入本发明的一种或多种酶突变体来去除接触镜片上的蛋白污物。
此发明的接触镜片洗涤组合物方案在下列实施例中阐明。
实施例91-94含酶接触镜片洗液实施例号组分91929394Val199Leu＋Ser210Asp0.01 0.5 0.1 2.0葡萄糖 50.0 50.0 50.0 50.0非离子表面活性剂(聚氧乙烯-聚氧 2.00 2.0 2.0 2.0丙烯共聚物)阴离子表面活性剂(聚氧乙烯烷基 1.00 1.0 1.0 1.0苯基醚硫酸酯(sulfricester)钠氯化钠 1.00 1.0 1.0 1.0硼砂0.30 0.3 0.3 0.3水 平衡至100％在实施例91-94中，表3-38所列举的枯草蛋白酶309突变体被突变体Val199Leu＋Ser210Asp所代替，结果基本相似。
由于有关本题的发明的具体方案已经描述了，对于本领域的技术人员来说，显而易见各种改变和修饰有关本题的发明，只要不脱离此发明的精神和范围，都是可以做到的。所附权利要求中意图覆盖所有此类本发明范围内的修饰。
序列表(1)一般信息(i)申请人PHILIP F.BRODE，IIIBOBBY L BARNETT
DONN N.RUBINCHCHANCHAL K.GHOSH(ii)发明题目吸附作用降低而水解作用增强的枯草蛋白酶309突变体(iii)序列个数2(iv)通信地址(A)收信人PROCTER & GAMBLE公司(B)街道11810 EAST MAM1 Rivtr ROAD(C)城市ROSS(D)州俄亥俄(E)国家美国(F)邮编 45061(v)机读形式(A)介质类型磁盘软盘(B)计算机IBM PC兼容机，(C)操作系统PC-DOS/US-DOS(D)软件Pate In Release#1.0，版#1.25(vi)目前申请数据(A)申请号(B)提交早期(C)分类(viii)律师代理信息(A)姓名CORSTANJE.BRAHM J.
(B)注册号34,804(C)代理人案号5233R(ix)电子通信信息(A)电话513-627-2858(B)传真513-627-0260(2)SEQ ID NO1序列信息(i)序列特征(A)长度269氨基酸(B)类型氨基酸
(C)链型单链(D)拓扑学线性(ii)分子类型蛋白质(xi)序列描述SEQ ID NO1Ala Gln Ser Val Pro Trp Gly Ile Ser Arg Val Gln Ala Pro Ala Ala1 5 10 15His Asn Arg Gly Leu Thr Gly Ser Gly Val Lys Val Ala Val Leu Asp20 25 30Thr Gly Ile Ser Thr His Pro Asp Leu Asn Ile Arg Gly Gly Ala Ser35 40 45Phe Val Pro Gly Glu Pro Ser Thr Gln Asp Gly Asn Gly His Gly Thr50 55 60His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu65 70 75 80Gly Val Ala Pro Ser Ala Glu Leu Tyr Ala Val Lys Val Leu Gly Ala85 90 95Ser Gly Ser Gly Ser Val Ser Ser Ile Ala Gln Gly Leu Glu Trp Ala100 105 110Gly Asn Asn Gly Met His Val Ala Asn Leu Ser Leu Gly Ser Pro Ser115 120 125Pro Ser Ala Thr Leu Glu Gln Ala Val Asn Ser Ala Thr Ser Arg Gly130 135 140Val Leu Val Val Ala Ala Ser Gly Asn Ser Gly Ala Gly ser Ile Ser145 150 155 160Tyr Pro Ala Arg Tyr Ala Asn Ala Met Ala Val Gly Ala Thr Asp Gln165 170 175Asn Asn Asn Arg Ala Ser Phe Ser Gln Tyr Gly Ala Gly Leu Asp Ile180 185 190Val Ala Pro Gly Val Asn Val Gln Ser Thr Tyr Pro Gly Ser Thr Tyr195 200 205Ala Ser Leu Asn Gly Thr Ser Met Ala Thr Pro His Val Ala Gly Ala210 215 220Ala Ala Leu Val Lys Gln Lys Asn Pro Ser Trp Ser Asn Val Gln Ile225 230 235 240Arg Asn His Leu Lys Asn Thr Ala Thr Ser Leu Gly Ser Thr Asn Leu245 250 255Tyr Gly Ser Gly Leu Val Asn Ala Glu Ala Ala Thr Arg260 265(B)类型氨基酸(C)拓扑学线性(ii)分子类型蛋白质(xi)序列描述SEQ ID NO2Ala Gln ser Val Pro Tyr Gly Val Ser Gln Ile Lys Ala Pro Ala Leu1 5 10 15His Ser Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp20 25 30Ser Gly Ile Asp Ser Ser His Pro Asp Leu Lys Val Ala Gly Gly Ala35 40 45Ser Met Val Pro Ser Glu Thr Asn Pro Phe Gln Asp Asn Asn Ser His50 55 60Gly Thr His Val Ala Gly Thr Val Ala Ala Leu Asn Asn Ser Ile Gly65 70 75 80Val Leu Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu85 90 95Gly Ala Asp Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu100 105 110Trp Ala Ile Ala Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly115 120 125Pro Ser Gly Ser Ala Ala Leu Lys Ala Ala Val Asp Lys Ala Val Ala130 135 140Ser Gly Val Val Val Val Ala Ala Ala Gly Asn Glu Gly Thr Ser Gly145 150 155 160Ser Ser Ser Thr Val Gly Tyr Pro Gly Lys Tyr Pro Ser Val Ile Ala165 170 175Val Gly Ala Val Asp Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Val180 185 190Gly Pro Glu Leu Asp Val Met Ala Pro Gly Val Ser Ile Gln Ser Thr195 200 205Leu Pro Gly Asn Lys Tyr Gly Ala Tyr Asn Gly Thr Ser Met Ala Ser210 215 220Pro His Val Ala Gly Ala Ala Ala Leu Ile Leu Ser Lys His Pro Asn225 230 235 240Trp Thr Asn Thr Gln Val Arg Ser Ser Leu Glu Asn Thr Thr Thr Lys245 250 255Leu Gly Asp Ser Phe Tyr Tyr Gly Lys Gly Leu Ile Asn Val Gln Ala260 265 270Ala Ala Gln27权利要求
1.一种具有修饰的枯草蛋白酶309野生型氨基酸序列的枯草蛋白酶309突变体，其特征在于修饰的氨基酸序列包含在如下一个或多个位点的替换193，194，195，196，197，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213，或214。其中a.当一个替换发生在193位点，用于替换的氨基酸为Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu；b.当一个替换发生在194位点，用于替换的氨基酸为His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。c.当一个替换发生在195位点，用于替换的氨基酸为Gly、Gln、Asn、Ser、Asp或Glu。d.当一个替换发生在196位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu。e.当一个替换发生在197位点，用于替换的氨基酸为Met、Cys、Ala、His、Pro、Gly、Gln、Ser、Asp或Glu。f.当一个替换发生在199位点，用于替换的氨基酸为Met、Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。g.当一个替换发生在200位点，用于替换的氨基酸为Asn、Ser或Ser。h.当一个替换发生在201位点，用于替换的氨基酸为Asp或Glu。i.当一个替换发生在202位点，用于替换的氨基酸为Pro、Gly、Gln、Asn、Ser、Asp或Glu。j.当一个替换发生在203位点，用于替换的氨基酸为Ile、Met、Ala、His、Pro、Gln、Asn、Ser或Asp。k.当一个替换发生在204位点，用于替换的氨基酸为Gly、Gln、Asn、Ser、Asp或Glu。l.当一个替换发生在205位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu。m.当一个替换发生在206位点，用于替换的氨基酸为Asp或Glu。n.当一个替换发生在207位点，用于替换的氨基酸为Pro、Gly、Gln、Asn、Ser、或Glu。o.当一个替换发生在208位点，用于替换的氨基酸为Leu、Ile、Val、Met、Cys、Ala、His、Pro、Gly、Gln、Asn、Asp或Glu。p.当一个替换发生在209位点，用于替换的氨基酸为His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。q.当一个替换发生在201位点，用于替换的氨基酸为Asp或Glu。r.当一个替换发生在211位点，用于替换的氨基酸为Ile、Val、Met、Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser或Asp；s.当一个替换发生在212位点，用于替换的氨基酸为Glu；t，当一个替换发生在213位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp、Ser或Glu；u.当一个替换发生在214位点，用于替换的氨基酸为Pro、Gly、Gln、Asn、Asp或Glu。这些突变使得枯草蛋白酶309突变体与野生型相比对于不溶性底物具有降低的吸附作用和增强的水解作用；其中优选地为i.当一个替换发生在200位点，用于替换的氨基酸为Ser；ii.当一个替换发生在205位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu；iii.当一个替换发生在208位点，用于替换的氨基酸为Leu、Ile、Val、Met、Cys、Ala、His、Pro、Gly、Gln、Asn、Asp或Gluiv.当一个替换发生在209位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu；v.当一个替换发生在210位点，用于替换的氨基酸为Asp或Glu。更优选的是当一个替换发生在如下一个或位点193，194，195，196，197，199，201，202，203，204，205，206，207，208，209，210，211，212，213或214，用于替换任何位点193，194，195，196，197，199，201，202，204，205，206，208，209，210，211，212，213或214的氨基酸为Asp或Glu；当一个替换发生在203位点，用于替换的氨基酸为Asp；而当一个替换发生在位点207或212一个或两个时，用于替换207或212的氨基酸为Glu。
2.权利要求1的带有单个氨基酸替换的枯草蛋白酶309突变体，其中替换为a.207位点Glu替换Thrb.210位点Glu替换Ser，或c.210位点Asp替换Ser。
3.一种具有修饰的枯草蛋白酶309野生型氨基酸序列的枯草蛋白酶309突变体，其特征在于修饰的氨基酸序列包含在如下三个或更多个位点的替换193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213或214。其中a.当一个替换发生在193位点，用于替换的氨基酸为Met、Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu；b.当一个替换发生在194位点，用于替换的氨基酸为His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。c.当一个替换发生在195位点，用于替换的氨基酸为Gly、Gln、Asn、Ser、Asp或Glu。d.当一个替换发生在196位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu。e.当一个替换发生在197位点，用于替换的氨基酸为Met、Cys、Ala、His、Pro、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。f.当一个替换发生在198位点，用于替换的氨基酸为Glu、Gln、Asp或Ser。g.当一个替换发生在199位点，用于替换的氨基酸为Met、Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。h.当一个替换发生在200位点，用于替换的氨基酸为Asn、Ser、Glu或Asp。i.当一个替换发生在201位点，用于替换的氨基酸为Asp或Glu。j.当一个替换发生在202位点，用于替换的氨基酸为Pro、Gly、Gln、Asn、Ser、Asp或Glu。k.当一个替换发生在203位点，用于替换的氨基酸为Leu、Ile、Val、Met、Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。l.当一个替换发生在204位点，用于替换的氨基酸为Gly、Gln、Asn、Ser、Asp或Glu。m.当一个替换发生在205位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu。n.当一个替换发生在206位点，用于替换的氨基酸为Asp或Glu。o.当一个替换发生在207位点，用于替换的氨基酸为Pro、Gly、Gln、Asn、Ser、Asp或Glu。p.当一个替换发生在208位点，用于替换的氨基酸为Leu、Ile、Val、Met、Cys、Thr、Ala、His、Pro、Gly、Gln、Asn、Asp或Glu。q.当一个替换发生在209位点，用于替换的氨基酸为His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。r.当一个替换发生在210位点，用于替换的氨基酸为Asp或Glu；s.当一个替换发生在211位点，用于替换的氨基酸为Ile、Val、Met、Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser、Glu或Asp；t，当一个替换发生在212位点，用于替换的氨基酸为Gln、Ser、Asp或Glu；u.当一个替换发生在213位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu；v.当一个替换发生在214位点，用于替换的氨基酸为Pro、Gly、Gln、Asn、Asp、Ser或Glu。这些突变使得枯草蛋白酶309突变体与野生型相比对于不溶性底物具有降低的吸附作用和增强的水解作用；优选地i.当一个替换发生在200位点，用于替换的氨基酸为Asn、Glu、Asp或Ser；ii.当一个替换发生在204位点，用于替换的氨基酸为Gly、Gln、Asn、Ser、Asp或Glu；iii.当一个替换发生在205位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Gluiv.当一个替换发生在208位点，用于替换的氨基酸为Leu、Ile、Val、Met、Cys、Ala、His、Pro、Gly、Gln、Asp或Glu；v.当一个替换发生在209位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu。vi.当一个替换发生在210位点，用于替换的氨基酸为Glu或Asp；
4.权利要求3的包含两个氨基酸替换的枯草蛋白酶309突变体，其中替换为a.207位点Glu替代Thr和210位点Glu替代Serb.197位点Leu替代Val和210位点Glu替代Ser；或c.199位点Leu替代Val和210位点Asp替代Ser；d.204位点Ala替代Pro和209位点Thr替代氨酸；e.208位点Phe替代Tyr和211位点Asn替代Leu；f.74位点Asp替代Asn和210位点Glu替代Ser；或g.207位点Glu替代Lys和210位点Glu替代Ala。
5.一种具有修饰的枯草蛋白酶309野生型氨基酸序列的枯草蛋白酶309突变体，其特征在于修饰的氨基酸序列包含在如下三个或更多个位点的替换193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213或214。在这些位点中a.当一个替换发生在193位点，用于替换的氨基酸为Met、Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu；b.当一个替换发生在194位点，用于替换的氨基酸为His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。c.当一个替换发生在195位点，用于替换的氨基酸为Gly、Gln、Asn、Ser、Asp或Glu。d.当一个替换发生在196位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu。e.当一个替换发生在197位点，用于替换的氨基酸为Met、Cys、Ala、His、Pro、Thr、Gly、Gln、Asn、Ser、Asp或Glu。f.当一个替换发生在198位点，用于替换的氨基酸为Glu、Gln、Asn、Asp或Ser。g.当一个替换发生在199位点，用于替换的氨基酸为Met、Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu、h.当一个替换发生在200位点，用于替换的氨基酸为Asn、Ser、Glu或Asp。i.当一个替换发生在201位点，用于替换的氨基酸为Asp或Glu。j.当一个替换发生在202位点，用于替换的氨基酸为Pro、Gly、Gln、Asn、Ser、Asp或Glu。k.当一个替换发生在203位点，用于替换的氨基酸为Leu、Ile、Val、Met、Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。l.当一个替换发生在204位点，用于替换的氨基酸为Gly、Gln、Asn、Ser、Asp或Glu。m.当一个替换发生在205位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu。n.当一个替换发生在206位点，用于替换的氨基酸为Asp或Glu。o.当一个替换发生在207位点，用于替换的氨基酸为Pro、Gly、Gln、Asn、Ser、Asp或Glu。p.当一个替换发生在208位点，用于替换的氨基酸为Leu、Ile、Val、Met、Cys、Thr、Ala、His、Pro、Gly、Gln、Asp或Glu。q.当一个替换发生在209位点，用于替换的氨基酸为His、Thr、Pro、Gly、Gln、Asn、Ser、Asp或Glu。r.当一个替换发生在210位点，用于替换的氨基酸为Asp或Glu；s.当一个替换发生在211位点，用于替换的氨基酸为Ile、Val、Met、Cys、Ala、His、Thr、Pro、Gly、Gln、Asn、Ser、Glu或Asp；t.当一个替换发生在212位点，用于替换的氨基酸为Gln、Ser、Asp或Glu；u.当一个替换发生在213位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu；v.当一个替换出生在214位点，用于替换的氨基酸为Pro、Gly、Gln、Asn、Asp、Ser或Glu。这些突变使得枯草蛋白酶309突变体与其野生型相比对不溶性底物具有降低的吸附作用和增强的水解作用；优选地i.当一个替换发生在200位点，用于替换的氨基酸为Asn、Glu、Asp或Ser；ii.当一个替换发生在204位点，用于替换的氨基酸为Gly、Gln、Asn、Ser、Asp或Glu；iii.当一个替换发生在205位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp和Glu；iv.当一个替换发生在208位点，用于替换的氨基酸为Leu、Ile、Val、Met、Cys、Ala、His、Pro、Gly、Gln、Asn、Asp或Glu；v.当一个替换发生在209位点，用于替换的氨基酸为Pro、Gln、Asn、Ser、Asp或Glu；以及vi.当一个替换发生在210位点，用于替换的氨基酸为Glu或Asp。
6.权利要求5中包含三个氨基酸替换的枯草蛋白酶309的突变体，其中替换为a.在200位点Glu替换Gln，205位点Ala替换Gly，以及210位点Glu替换Ser；b.199位点Leu替换Val，204位点Ala替换Pro，以及207位点Glu替代Thr；c.74位点Asp替代Asn，200位点Glu替代Gln，以及210位点Glu替换Ser；或d.204位点Glu替代Gln，207位点Glu替代Thr，和210位点Glu替代Ser。
7.权利要求5的包含如下4个替换的枯草蛋白酶309突变体；204位点Ala替代Pro，207位点Glu替代Thr，210位点Glu替代Ser，和74位点Asp替代Asn。
8.权利要求5的包含下列5个替换的枯草蛋白酶突变体199位Leu替代Ile、204位点Ala替代Pro，207位点Glu替代Lys，210位点Glu替代Ala，和74位点Asp替代Asn。
9.一种具有修饰的枯草蛋白酶309野生型氨基酸序列的枯草蛋白酶309突变体，野生氨基酸序列包含第1环状区，第2环状区，第3环状区，第4环状区和第5环状区；其特征在于修饰的氨基酸序列在一个或多个环状区的位点包含一个或多个替换，其中A.当一个替换发生在第1环状区，替换发生在如下一个或多个位点57，58，59，60，61，63或64，其中a.当一个替换发生在57位点，用于替换的氨基酸为Asn、Asp、Glu或Ser；b.当一个替换发生在58位点，用于替换的氨基酸为Glu；c.当一个替换发生在59位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser；d.当一个替换发生在60位点，用于替换的氨基酸为Asp、Gln、Glu和Ser；e.当一个替换发生在61位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro和Ser；f.当一个替换发生在63位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro和Ser；g.当一个替换发生在64位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro和Ser；B.当一个替换发生在第2环状区，替换发生在如下一个或多个位点93，94，95，96，97，98，99，100，101，102，103，104或105；其中a.当一个替换发生在93位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr；b.当一个替换发生在94位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val；c.当一个替换发生在95位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser；d.当一个替换发生在96位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。e.当一个替换发生在97位点，用于替换的氨基酸为Asp或Glu。f.当一个替换发生在98位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。g.当一个替换发生在99位点，用于替换的氨基酸为Asp或Glu；h.当一个替换发生在100位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser；i.当一个替换发生在101位点，用于替换的氨基酸为Asp或Glu。j.当一个替换发生在102位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr；k.当一个替换发生在103位点，用于替换的氨基酸为Asp或Glu。l.当一个替换发生在104位点，用于替换的氨基酸为Asp或Glu。m.当一个替换发生在105位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Leu、Met、Pro、Ser、Thr或Val；C.当一个替换发生在第3环状区时，替换发生在如下一个或多个位点124，125，126，127，128，129，130或131；其中a.当一个替换发生在124位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val；b.当一个替换发生在125位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser；c.当一个替换发生在126位点，用于替换的氨基酸为Asp或Glu；d.当一个替换发生在127位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser；e.当一个替换发生在128位点，用于替换的氨基酸为Asp或Glu。f.当一个替换发生在129位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser。g.当一个替换发生在130位点，用于替换的氨基酸为Asp或Glu；h.当一个替换发生在131位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr；D.当一个替换发生在第四环状区，替换发生在如下一个或多个位点152，153，154，155，156，157，158，159，160或161；其中a.当一个替换发生在152位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser；b.当一个替换发生在153位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser；c.当一个替换发生在154位点，用于替换的氨基酸为Asp或Glu；d.当一个替换发生在155位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser；e.当一个替换发生在156位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr；f.当一个替换发生在157位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser。g.当一个替换发生在158位点，用于替换的氨基酸为Asp或Glu；h.当一个替换发生在159位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Leu、Met、Pro、Ser、Thr或Val；i.当一个替换发生在160位点，用于替换的氨基酸为Asp或Glu。j.当一个替换发生在161位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Ser或Val；E.当一个替换发生在第五环状区，替换发生在如下一个或多个位点181，182，183，184或185，其中a.当一个替换发生在181位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr；b.当一个替换发生在182位点，用于替换的氨基酸为Asp、或Glu；c.当一个替换发生在183位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Ser、Thr、Tyr或Val；d.当一个替换发生在184位点，用于替换的氨基酸为Asp或Glu；e.当一个替换发生在185位点，用于替换的氨基酸为Asp、Asp、Glu或Ser；这些替换使得枯草蛋白酶309突变体与野生型相比具有降低的吸附作用和增强的水解作用；
10.权利要求9中的枯草蛋白酶309突变体，其中枯草蛋白酶309野生型氨基酸序列进一步包含一个第六环状区，其中修饰的氨基酸序列在第六环状区进一步包括一个或多个替换；这些存在于第六环状区的替换发生在如下一个或多个位点193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213或214；其中a.当一个替换发生在193位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr；b.当一个替换发生在194位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr；c.当一个替换发生在195位点，用于替换的氨基酸为Asn、Asp、Gln、Glu或Ser；d.当一个替换发生在196位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser；e.当一个替换发生在197位点，用于替换的氨基酸为Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr；f.当一个替换发生在198位点，用于替换的氨基酸为Asp、Gln、Glu或Ser。g.当一个替换发生在199位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr；h.当一个替换发生在200位点，用于替换的氨基酸为Asn、Asp、Glu或Ser；i.当一个替换发生在201位点，用于替换的氨基酸为Asp或Glu。j.当一个替换发生在202位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、 Pro或Ser；k.当一个替换发生在203位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Ser、Thr或Val；l.当一个替换发生在204位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly或Ser；m.当一个替换发生在205位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser；n.当一个替换发生在206位点，用于替换的氨基酸为Asp或Glu。o.当一个替换发生在207位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser；p.当一个替换发生在208位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Ser、Thr或Val；q.当一个替换发生在209位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr；r.当一个替换发生在210位点，用于替换的氨基酸为Asp或Glu；s.当一个替换发生在211位点，用于替换的氨基酸为Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val；t.当一个替换发生在212位点，用于替换的氨基酸为Asn、Gln、Glu或Ser；u.当一个替换发生在213位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Pro或Ser；v.当一个替换发生在214位点，用于替换的氨基酸为Asn、Asp、Gln、Glu、Gly、Pro或Ser；
11.一种选自硬表面洗涤组合物，餐具洗涤组合物，口腔清洁组合物、假牙清洗组合物、接触镜片洗涤组合物和织物洗涤组合物的洗涤组合物，其特征在于此洗涤组合物包括权利要求1-11中任一项的枯草蛋白酶309突变体和一种洗涤组合物载体。
12.权利要求1-11任一项的枯草蛋白酶309突变体，其中修饰的氨基酸序列进一步包括在74位点His或Asp对Asn的替换。
13.权利要求11中的洗涤组合物，其中洗涤组合物是一种硬表面洗涤组合物。
14.权利要求11中的洗涤组合物，其中洗涤组合物为一种织物洗涤组合物，组合物优选为液体形式。
15.一种编码权利要求1-11任一项的枯草蛋白酶309突变体的309基因DNA序列。
全文摘要
本发明涉及具有修饰的野生型枯草蛋白酶(309)氨基酸序列的枯草蛋白酶突变体(309)，野生型氨基酸序列包括第一环状区，第二环状区，第三环状区，第四环状区，第五环状区和第六环状区。其中修饰后的氨基酸序列在一个或多个环状区比野生型枯草蛋白酶(309)在特定的鉴定位点上有不同的氨基酸(即替换)，从而使枯草蛋白酶(309)突变体比野生型枯草蛋白酶相比对不溶性底物有降低的吸附作用和增强的水解作用。本发明还涉及编码这类枯草蛋白酶(309)突变体的DNA序列。本发明还涉及包含这类枯草蛋白酶(309)突变体的用于洗涤各类表面的组合物。
文档编号G06Q40/00GK1151187SQ95193744
公开日1997年6月4日 申请日期1995年4月18日 优先权日1994年5月2日
发明者P·F·布罗德三世, B·L·巴尼特, D·N·鲁宾, C·K·高希 申请人:普罗格特-甘布尔公司