来自嗜热棒状杆菌的氨基酸生物合成途径的抗热酶的基因的制作方法

专利名称：来自嗜热棒状杆菌的氨基酸生物合成途径的抗热酶的基因的制作方法
技术领域：
本发明涉及到抗热酶的基因，特别是涉及到Corynebacteriumthermoaminogenes的L-氨基酸，例如L-谷氨酸的生物合成和吸收系统的基因，该菌类为一种嗜热棒状杆菌。
背景工艺
诸如L-谷氨酸这样的L-氨基酸的生产的当今主流为利用棒状杆菌的发酵生产。关于L-氨基酸的发酵生产，已经通过培育具有优良产量的菌株以及开发发酵工艺来尽力降低成本。尽管对于实现成本降低的传统努力主要针对于获得高产率，除了作为发酵成本所涉及因素的原材料之外，对在培养期间所产生的发酵热量进行冷却的能量也不可忽视。这就是说，考虑到用于发酵的为普通的微生物，由于微生物自身在发酵期间所产生的发酵热量会引起温度的升高并且发酵所需的酶由此可能会失活或者生产性细菌可能会因此而被杀死。因此，在发酵期间对培养基的冷却是必需的。由此，为降低冷却成本，多年以来一直在进行对高温发酵的研究。进一步，如果高温发酵成为可能，反应的速度也可能会提高。但是，对于L-氨基酸发酵，有效的高温培养迄今为止尚未实现。
Corynebacterium thermoaminogenes为一种归类于棒状细菌的细菌，它类似于谷氨酸棒状杆菌(乳发酵短杆菌(Brevibacteriumlactofermentum))，该菌株通常被用于L-氨基酸的发酵。然而，据显示其最佳生长温度为37-43℃，该温度高于谷氨酸棒状杆菌的最佳生长温度，即30-35℃，并且对于L-谷氨酸生产该最佳温度为42-45℃，该温度明显地偏向高温区域(Japanese Patent Laid-open(Kokai)No.63-240779/1988)。
与此同时，用于通过向其中引入一种L-氨基酸合成系统酶来增强棒状杆菌属和短杆菌属的L-氨基酸生产能力的工艺已被开发，该酶来自大肠杆菌或者谷氨酸棒状杆菌。这样的酶的例子包括，例如，柠檬酸合成酶(日本专利出版物(Kokoku)No.7-121228/1995)，该酶为L-谷氨酸生物合成途径的一种酶，这样的例子还包括谷氨酸脱氢酶(Japanese Patent Laid-open No.61-268185/1986)、异柠檬酸脱氢酶、乌头酸水合酶(Japanese Patent Laid-open No.63-214189)等等。
但是，任何来自嗜热棒状杆菌的L-氨基酸合成酶以及编码它们的基因均未被报道。
发明披露
本发明的一个目的为提供编码来自Corynebacteriumthermoaminogenes的酶的基因，在优选的情况下提供在高于谷氨酸棒状杆菌的对应温度下发挥功能的酶。
本发明的发明者进行了广泛的研究以期达到上述的目的。其结果，他们成功地分离了编码Corynebacterium thermoaminogenes的氨基酸生物合成途径的酶的基因或者编码参与细胞摄入氨基酸的蛋白的基因，并且由此实现了本发明。
简而言之，本发明提供了以下的方面。
(1)一种蛋白质，该蛋白质具有SEQ ID NO2的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO2的氨基酸序列，该蛋白质具有异柠檬酸裂合酶活性并且在50℃下进行5分钟的热处理后表现出30％或者更高的残余活性。
(2)一种蛋白质，该蛋白质具有SEQ ID NO4的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO4的氨基酸序列，该蛋白质涉及到来自Corynebacteriumthermoaminogenes的脂酰辅酶A羧化酶活性。
(3)一种蛋白质，该蛋白质具有SEQ ID NO6的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO6的氨基酸序列，该蛋白质具有来自Corynebacteriumthermoaminogenes的DtsR活性。
(4)一种蛋白质，该蛋白质具有SEQ ID NO8的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO8的氨基酸序列，该蛋白质具有来自Corynebacteriumthermoaminogenes的DtsR活性。
(5)一种蛋白质，该蛋白质具有SEQ ID NO10的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO10的氨基酸序列，该蛋白质在60℃所表现出的磷酸果糖激酶活性同在30℃相比与之等效或者比后者水平更高。
(6)一种蛋白质，该蛋白质具有SEQ ID NO94的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO94的氨基酸序列，该蛋白质具有赋予Corynebacteriumthermoaminogenes蔗糖同化能力的活性。
(7)一种蛋白质，该蛋白质具有SEQ ID NOS17-20的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NOS17-20的氨基酸序列，该蛋白质具有的功能涉及到谷氨酸的摄取并且它来自于Corynebacteriumthermoaminogenes。
(8)一种蛋白质，该蛋白质具有SEQ ID NO22的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO22的氨基酸序列，该蛋白质具有来自Corynebacteriumthermoaminogenes的丙酮酸脱氢酶活性。
(9)一种蛋白质，该蛋白质具有SEQ ID NO24的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO24的氨基酸序列，该蛋白质具有来自Corynebacteriumthermoaminogenes的丙酮酸羧化酶活性。
(10)一种蛋白质，该蛋白质具有SEQ ID NO26的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO26的氨基酸序列，该蛋白质具有磷酸烯醇丙酮酸羧化酶活性并且在45℃下进行5分钟的热处理后表现出50％或者更高的残余活性。
(11)一种蛋白质，该蛋白质具有SEQ ID NO28的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO28的氨基酸序列，该蛋白质具有顺乌头酸酶活性并且在50℃下进行3分钟的热处理后表现出30％或者更高的残余活性。
(12)一种蛋白质，该蛋白质具有SEQ ID NO30的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO30的氨基酸序列，该蛋白质具有异柠檬酸脱氢酶活性并且在45℃下进行10分钟的热处理后表现出50％或者更高的残余活性。
(13)一种蛋白质，该蛋白质具有SEQ ID NO32的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO32的氨基酸序列，该蛋白质具有来自Corynebacterium thermoaminogenes的二氢硫辛酰胺脱氢酶活性。
(14)一种蛋白质，该蛋白质具有SEQ ID NO34的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO34的氨基酸序列，该蛋白质具有2-酮戊二酸脱氢酶活性并且在50℃下进行10分钟的热处理后表现出30％或者更高的残余活性。
(15)一种蛋白质，该蛋白质具有SEQ ID NO80的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO80的氨基酸序列，该蛋白质在42℃所表现出的谷氨酸脱氢酶活性同在37℃相比与之等效或者比后者水平更高。
(16)一种蛋白质，该蛋白质具有SEQ ID NO90的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO90的氨基酸序列，该蛋白质在37℃所表现出的柠檬酸合成酶活性同在23℃相比与之等效或者比后者水平更高。
(17)一种DNA，该DNA编码的蛋白质具有SEQ ID NO2的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO2的氨基酸序列，并且该蛋白质具有异柠檬酸裂合酶活性。
(18)(17)中的DNA，该DNA为下列的(a1)或者(b1)中所定义的DNA
(a1)一种含有序列表中的SEQ ID NO1的核苷酸序列的DNA，
(b1)一种DNA，该DNA同序列表中的SEQ ID NO1的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有异柠檬酸裂合酶活性的蛋白质。
(19)一种DNA，该DNA编码的蛋白质具有SEQ ID NO4的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO4的氨基酸序列，并且该蛋白质涉及到脂酰辅酶A羧化酶活性。
(20)(19)中的DNA，该DNA为下列的(a2)或者(b2)中所定义的DNA
(a2)一种含有序列表中的SEQ ID NO3的核苷酸序列的DNA，
(b2)一种DNA，该DNA同序列表中的SEQ ID NO3的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种涉及到脂酰辅酶A羧化酶活性的蛋白质。
(21)一种DNA，该DNA编码的蛋白质具有SEQ ID NO6的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO6的氨基酸序列，并且该蛋白质具有DtsR活性。
(22)(21)中的DNA，该DNA为下列的(a3)或者(b3)中所定义的DNA
(a3)一种含有序列表中的SEQ ID NO5的核苷酸序列的DNA，
(b3)一种DNA，该DNA同序列表中的SEQ ID NO5的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有DtsR活性的蛋白质。
(23)一种DNA，该DNA编码的蛋白质具有SEQ ID NO8的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO8的氨基酸序列，并且该蛋白质具有DtsR活性。
(24)(23)中的DNA，该DNA为下列的(a4)或者(b4)中所定义的DNA
(a4)一种含有序列表中的SEQ ID NO7的核苷酸序列的DNA，
(b4)一种DNA，该DNA同序列表中的SEQ ID NO7的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有DtsR活性的蛋白质。
(25)一种DNA，该DNA编码的蛋白质具有SEQ ID NO10的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO10的氨基酸序列，并且该蛋白质具有磷酸果糖激酶活性。
(26)(25)中的DNA，该DNA为下列的(a5)或者(b5)中所定义的DNA
(a5)一种含有序列表中的SEQ ID NO9的核苷酸序列的DNA，
(b5)一种DNA，该DNA同序列表中的SEQ ID NO9的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有磷酸果糖激酶活性的蛋白质。
(27)一种DNA，该DNA编码的蛋白质具有SEQ ID NO93的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO93的氨基酸序列，并且该蛋白质具有蔗糖酶(invertase)活性。
(28)(27)中的DNA，该DNA为下列的(a6)或者(b6)中所定义的DNA
(a6)一种含有序列表中的SEQ ID NO93的核苷酸序列的DNA，
(b6)一种DNA，该DNA同序列表中的SEQ ID NO93的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有蔗糖酶活性的蛋白质。
(29)一种DNA，该DNA编码的蛋白质具有SEQ ID NOS17-20的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NOS17-20的氨基酸序列，并且该蛋白质具有参与谷氨酸摄入的功能。
(30)(29)中的DNA，该DNA为下列的(a7)或者(b7)中所定义的DNA
(a7)一种含有序列表中的SEQ ID NO16的核苷酸序列的DNA，
(b7)一种DNA，该DNA同序列表中的SEQ ID NO16的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有参与谷氨酸摄入的功能的蛋白质。
(31)一种DNA，该DNA编码的蛋白质具有SEQ ID NO22的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO22的氨基酸序列，并且该蛋白质具有丙酮酸脱氢酶活性。
(32)(31)中的DNA，该DNA为下列的(a8)或者(b8)中所定义的DNA
(a8)一种含有序列表中的SEQ ID NO21的核苷酸序列的DNA，
(b8)一种DNA，该DNA同序列表中的SEQ ID NO21的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有丙酮酸脱氢酶活性的蛋白质。
(33)一种DNA，该DNA编码的蛋白质具有SEQ ID NO24的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO24的氨基酸序列，并且该蛋白质具有丙酮酸羧化酶活性。
(34)(33)中的DNA，该DNA为下列的(a9)或者(b9)中所定义的DNA
(a9)一种含有序列表中的SEQ ID NO23的核苷酸序列的DNA，
(b9)一种DNA，该DNA同序列表中的SEQ ID NO23的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有丙酮酸羧化酶活性的蛋白质。
(35)一种DNA，该DNA编码的蛋白质具有SEQ ID NO26的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO26的氨基酸序列，并且该蛋白质具有磷酸烯醇丙酮酸羧化酶活性。
(36)(35)中的DNA，该DNA为下列的(a10)或者(b10)中所定义的DNA
(a10)一种含有序列表中的SEQ ID NO25的核苷酸序列的DNA，
(b10)一种DNA，该DNA同序列表中的SEQ ID NO25的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有磷酸烯醇丙酮酸羧化酶活性的蛋白质。
(37)一种DNA，该DNA编码的蛋白质具有SEQ ID NO28的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO28的氨基酸序列，并且该蛋白质具有顺乌头酸酶活性。
(38)(37)中的DNA，该DNA为下列的(a11)或者(b11)中所定义的DNA
(a11)一种含有序列表中的SEQ ID NO27的核苷酸序列的DNA，
(b11)一种DNA，该DNA同序列表中的SEQ ID NO27的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有顺乌头酸酶活性的蛋白质。
(39)一种DNA，该DNA编码的蛋白质具有SEQ ID NO30的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO30的氨基酸序列，并且该蛋白质具有异柠檬酸脱氢酶活性。
(40)(39)中的DNA，该DNA为下列的(a12)或者(b12)中所定义的DNA
(a12)一种含有序列表中的SEQ ID NO27的核苷酸序列的DNA，
(b12)一种DNA，该DNA同序列表中的SEQ ID NO27的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有异柠檬酸脱氢酶活性的蛋白质。
(41)一种DNA，该DNA编码的蛋白质具有SEQ ID NO32的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO32的氨基酸序列，并且该蛋白质具有二氢硫辛酰胺脱氢酶活性。
(42)(41)中的DNA，该DNA为下列的(a13)或者(b13)中所定义的DNA
(a13)一种含有序列表中的SEQ ID NO31的核苷酸序列的DNA，
(b13)一种DNA，该DNA同序列表中的SEQ ID NO31的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有二氢硫辛酰胺脱氢酶活性的蛋白质。
(43)一种DNA，该DNA编码的蛋白质具有SEQ ID NO34的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO34的氨基酸序列，并且该蛋白质具有2-酮戊二酸脱氢酶活性。
(44)(43)中的DNA，该DNA为下列的(a14)或者(b14)中所定义的DNA
(a14)一种含有序列表中的SEQ ID NO33的核苷酸序列的DNA，
(b14)一种DNA，该DNA同序列表中的SEQ ID NO33的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码一种具有2-酮戊二酸脱氢酶活性的蛋白质。
(45)一种DNA，该DNA编码的蛋白质具有SEQ ID NO80的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO80的氨基酸序列，并且该蛋白质在42℃所表现出的谷氨酸脱氢酶活性同在37℃相比与之等效或者比后者水平更高。
(46)(45)中的DNA，该DNA为下列的(a15)或者(b15)中所定义的DNA
(a15)一种含有序列表中的SEQ ID NO79的核苷酸序列的DNA，
(b15)一种DNA，该DNA同序列表中的SEQ ID NO79的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码的蛋白质在42℃所表现出的谷氨酸脱氢酶活性同在37℃相比与之等效或者比后者水平更高。
(47)一种DNA，该DNA编码的蛋白质具有SEQ ID NO90的氨基酸序列或者为具有一个或多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO90的氨基酸序列，并且该蛋白质在37℃所表现出的柠檬酸合成酶活性同在23℃相比与之等效或者比后者水平更高。
(48)(47)中的DNA，该DNA为下列的(a16)或者(b16)中所定义的DNA
(a15)一种含有序列表中的SEQ ID NO89的核苷酸序列的DNA，
(b15)一种DNA，该DNA同序列表中的SEQ ID NO89的核苷酸序列或者同基于该核苷酸序列的引物可在严格的条件下杂交，并且该DNA编码的蛋白质在37℃所表现出的柠檬酸合成酶活性同在23℃相比与之等效或者比后者水平更高。
(49)一种用于产生L-氨基酸的方法，该方法包括培养一种微生物体以在培养基中产生和积累L-氨基酸，并且从该培养基中收集L-氨基酸，该微生物体中引入了(17)到(48)中任意一种的DNA。
术语“本发明的DNA”在此后被用于指上述的DNA之一或者全部。
在后文中将对本发明进行详细解释。
表1所示为本发明的DNA的核苷酸序列，基因的名称以及本发明的DNA所编码的蛋白质。
SEQ ID NOS3、23、25、31和33的开放读码框架(ORF)以及SEQ ID NO16的第四个ORF均由GTG开始。尽管由这些GTG所编码的氨基酸在序列表中被示为缬氨酸，但它们有可能为甲硫氨酸。
SEQ ID NO16的序列含有4个ORF，它们从5’末端按顺序对应于gluA、gluB、gluC和gluD。
上述的DNA序列分离自Corynebacterium thermoaminogenesAJ12310菌株(FERM BP-1542)的染色体DNA。然而，在SEQ ID NOS11和13中所示的DNA序列分别分离自Corynebacteriumthermoaminogenes AJ12340菌株(FERM BP-1539)和AJ12309菌株(FERM BP-1541)，它们分别具有蔗糖酶活性以及蔗糖同化特性，这是因为AJ12310菌株不具有蔗糖酶活性以及蔗糖同化特性，并且分离自该菌株的scrB基因不具有任何开放读码框架。
Corynebacterium thermoaminogenes AJ12310菌株(亦称为YS-314菌株)以及AJ12309菌株(亦称为YS-155菌株)于1987年3月13日保存于National Institute of Bioscience and Human-Technology，Agency of Industrial Science and Technology，Ministry of International Trade and Industry(1-3，Higashi1-chome，Tsukubashi，Ibaraki-ken，Japan，邮编305-8566)并且分别被分配以保存号FERM P-9246和FERM P-9245。随后，它们于1987年10月27日根据布达佩斯协定移送至国际保存(internationaldespositon)并且分别被分配以保存号FERM BP-1542和FERM BP-1541。
AJ12340菌株(亦称为YS-40菌株)于1987年3月10日保存于National Institute of Bioscience and Human-Technology，Agencyof Industrial Science and Technology，Ministry ofInternational Trade and Industry(1-3，Higashi 1-chome，Tsukubashi，Ibaraki-ken，Japan，邮编305-8566)并且被分配以保存号FERM P-9277。随后，它于1987年10月27日根据布达佩斯协定移送至国际保存并且分别被分配以保存号FERM BP-1539。
SEQ ID NOS11、13和15中所示的核苷酸序列为scrB的部分序列，并且SEQ ID NOS11和13编码在SEQ ID NOS12和14中所示的蔗糖酶的部分氨基酸序列。
含有一种靶基因的一个部分片段的DNA序列，可通过对已经报道的多种微生物中的该靶基因的核苷酸序列进行比较，以对高度保守的核苷酸序列进行选择，并且使用基于该区域的核苷酸序列而设计的引物以及使用Corynebacterium thermoaminogenes的染色体DNA作为模板进行PCR反应而获得，这样的微生物的例子如乳发酵短杆菌。进一步，通过使用所获得的DNA片段或者基于该片段序列而制备的探针进行杂交而对Corynebacterium thermoaminogenes的染色体DNA文库进行筛选，可以获得含有全长的该基因的DNA片段。一种含有该全长基因的DNA片段也可以通过使用所获得的该基因的部分片段进行基因组步移而获得。该基因组步移可通过使用一种商业化的试剂盒而进行，例如Takara LA PCR体外克隆试剂盒(Takara Shuro制造)。
例如，编码谷氨酸脱氢酶的DNA(后文中该DNA亦称为“gdh”，该酶亦称为“GDH”)的部分序列可通过PCR(聚合酶链反应)而从Corynebacterium thermoaminogenes，例如，从Corynebacteriumthermoaminogenes AJ12310菌株中获得，该PCR使用染色体DNA作为模板并且使用具有序列表中的SEQ ID NOS77和78中的核苷酸序列的引物而进行。进一步，完整的gdh基因可通过使用所获得的部分片段进行的基因组步移来获得。
进一步，编码柠檬酸合成酶的DNA(后文中该DNA亦称为“gltA”，该酶亦称为“CS”)的部分序列可通过PCR(聚合酶链反应)而从Corynebacterium thermoaminogenes，例如，从Corynebacteriumthermoaminogenes AJ12310菌株中获得，该PCR使用染色体DNA作为模板并且使用具有序列表中的SEQ ID NOS83和84中的核苷酸序列的引物而进行。进一步，完整的gltA基因可通过使用所获得的部分片段进行的基因组步移来获得。
上述的引物的核苷酸序列基于一种区域中的核苷酸序列进行设计，所述的区域含有多种微生物已经报道的gdh基因或者gltA基因中的高度保守的核苷酸序列，该区域通过对这些基因进行比较而发现。
对于编码其它酶的DNA序列，编码这些酶的部分片段可使用在表1中提及的引物以类似的方式获得，并且全长的该基因可通过使用所获得的该部分片段而获得。
尽管本发明的DNA可根据上述方法而获得，但该DNA还可以从Corynebacterium thermoaminogenes的一个染色体DNA文库中通过使用一种寡核苷酸作为探针而获得，该寡核苷酸的制备基于本发明的DNA的核苷酸序列。
用于染色体DNA的制备的方法、染色体DNA文库的结构、杂交、PCR、质粒DNA的制备、DNA的消化和连接、转化等等的描述见Sambrook，J.，Fritsch，E.F.，Maniatis，T.，Molecular Cloning，ColdSpring Harbor Laboratory Press，1.21(1989)。进一步，基因组步移可通过使用商业化的试剂盒而进行，例如Takara LA PCR体外克隆试剂盒(Takara Shuro制造)。
用于获得本发明的DNA的具体方法将在下文中进行解释。
首先，以一种适当的内切酶，例如Sau3AI，对Corynebacteriumthermoaminogenes的染色体DNA进行消化，并且通过琼脂糖凝胶电泳进行分离以获得一种约4到6kb的DNA片段。将所获得的DNA片段插入克隆载体，例如pHSG399，并且以所获得的重组质粒对大肠杆菌进行转化以产生染色体DNA的质粒文库。
另外，生产一些引物以用于通过PCR从质粒文库中选择含有靶基因的克隆。这些引物的设计基于来自多种微生物体的对应于目的基因的保守氨基酸区域。在这些引物的设计中，一些引物组出于棒状杆菌的密码子使用性的考虑而进行设计。
随后，为检测所产生引物的合理性，使用这些引物并且使用Corynebacterium thermoaminogenes的染色体DNA作为模板而进行了PCR。进一步，使用可获得扩增片段的引物作为筛选引物并且使用由该质粒文库中制备的重组质粒作为模板来进行PCR，以对含有靶DNA片段的克隆进行选择。这一操作过程的快速进行可通过对每个含有数十个转化子菌株的批次进行初级筛选，并且对获得了扩增片段的批次进行菌落PCR来进行次级筛选而完成。所扩增基因的片段长度示于表2到7。
通过从根据上法所选出的转化子中制备重组DNA、使用双脱氧终止法测定该插入片段的核苷酸序列并且以该核苷酸序列同已知基因序列进行比较，来证实是否根据上述方法所选出的转化子含有该靶基因。
当所获得的DNA片段含有该靶基因的一部分时，通过基因组步移来获得被缺失的部分。
本发明的DNA可能编码一个含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的蛋白质，只要所编码的蛋白质具有其原功能便可。“多个”所指的数量可根据蛋白质三维结构中的位置或者氨基酸残基的种类而变化。但是，在通常的情况下这样的一种蛋白质优选地表现出同蛋白质全部氨基酸序列具有30到40％或更多的同源性，在更优选的情况下为55到65％或更多。更具体的情况下，术语“多个”所示的数目为2到数百，在优选的情况下为2到数十，更优选的情况下为2到10。
核苷酸和氨基酸序列的分析通过，例如，由Lipman和Peason所开发出的方法来进行(Science，227，1435-1441，1985)，该方法使用商业化的软件进行，例如Genetyx-Mac计算机程序(SoftwareDevelopment Co.，Tokyo，Japan)。
GDH可能同组成GDH的全部氨基酸序列表现出40％到80％或更多的同源性，在优选的情况下为80到90％或更多，并且在42℃下表现出同在37℃下等同或者更高的活性。在这一情况下，该术语“多个”指的数量为2到30，在优选的情况下为2到50，更优选的情况下为2到10。
CS可能同组成CS的全部氨基酸序列表现出40％到80％或更多的同源性，在优选的情况下为80到90％或更多，并且在37℃下表现出同在23℃下等同或者更高的活性。在这一情况下，该术语“多个”指的数量为2到300，在优选的情况下为2到50，更优选的情况下为2到10。
一种编码与上述源蛋白质基本相同的蛋白的DNA，可通过例如，通过诸如定点突变这样的方法对核苷酸进行修饰以使一个特定位置的一个或多个氨基酸残基参与替换、缺失、插入、添加或者反转来获得。上述的经修饰的DNA还可能通过传统的已知突变处理来获得。该突变处理包括一种用于在体外处理编码靶基因的DNA的方法，例如，以羟胺进行，还包括一种以紫外照射或者通常用于突变处理的突变试剂处理携带有编码靶基因的DNA的微生物的方法，例如，处理属于大肠杆菌属的细菌，该突变试剂的例子如N-甲基-N’-硝基-N-亚硝基胍(NTG)和亚硝酸。
上述的核苷酸的替换、缺失、插入、添加或反转还包括由于Corynebacterium thermoaminogenes的菌株差异所造成的突变体或变体等。
一种编码与上述源蛋白质基本相同的蛋白的DNA，可通过在适当的细胞中表达有突变的DNA并检测该被表达产物蛋白的活性或功能而获得。该编码与源蛋白质基本相同的蛋白的DNA还可通过，例如，从编码具有突变的蛋白质的DNA中或从携带该DNA的细胞中分离一种可同具有某种核苷酸序列的DNA或者探针在严格条件下杂交的DNA来进行，该核苷酸序列为在表1中所提及的序列号的序列或其编码区域，该探针基于该核苷酸序列而设计，所分离的DNA编码的蛋白质具有该蛋白质原初具有的功能。该活性在优选的情况下对于GDH指42℃下的酶活性或者对于CS指37℃下的酶活性。
通过使用适当的引物而进行的PCR，上述的探针可从任意一个具有表1所示的序列号的核苷酸序列的DNA中或者具有任一该核苷酸序列的DNA制备。
“严格条件”在此处指的是一种条件，在该条件下形成所谓的特异性杂交体并且不能形成非特异性杂交体。使用任何数值来清楚地表述该条件都是很困难的。但是，例如，该严格条件包括一种条件，在该条件下具有高同源性的DNA，例如具有不低于50％的同源性的DNA之间可以互相杂交，并且在该条件下所具有的同源性低于上值的DNA之间不能杂交。或者，严格条件的例子为一种条件，在该条件下DNA的相互杂交所处的盐浓度对应于Southern杂交的普通洗涤条件，即，60℃、1×SSC、0.1％的SDS，在优选的情况下为0.1×SSC、0.1％的SDS。
可在上述条件下进行杂交的基因，包括在其中产生了终止密码子的基因以及由于活性位点的突变而无活性的基因。但是，这样的基因可以通过将其同商业化的活性表达载体相连接并且测量其活性或功能而被简单的除去。
对应于本发明的各DNA的蛋白质可通过在适当的宿主-载体系统中表达该DNA而产生。
作为用于基因表达的宿主，多种原核细胞可被提及，包括乳发酵短杆菌(谷氨酸棒状杆菌)、诸如Corynebacterium thermoaminogenes这样的棒状杆菌、大肠杆菌、枯草芽孢杆菌等等，以及多种真核细胞，包括酿酒酵母、动物细胞和植物细胞。这些细胞中，原核细胞，特别是棒状杆菌和大肠杆菌是优选的。
如果本发明的DNA同一种可在大肠杆菌和/或棒状杆菌细胞中自行复制的载体DNA连接而形成了重组DNA，并且该重组子DNA被引入了一种大肠杆菌细胞，随后的步骤便会变得简单。在大肠杆菌细胞中自行复制的载体在优选的情况下为一种可在宿主细胞中自行复制的质粒载体，该质粒载体的例子包括pUC19、pUC18、pBR322、pHSG299、pHSG399、pHSG398、RSF1010等。
作为可在棒状杆菌细胞中自行复制的载体，可提及的有pAM330(参考Japanese Patent Laid-open No.58-67699/1983)、pHM1519(参考Japanese Patent Laid-open No.58-77895/1983)等。而且，如果将具有制造可在棒状杆菌细胞中自行复制的质粒的能力的DNA片段从这些质粒中取出并插入上述用于大肠杆菌的载体中，所产生的载体可被用作为所谓的穿梭载体，该载体在大肠杆菌和棒状杆菌中均可自行复制。
这样的穿梭载体的例子包括下文提及的载体。所示的还有携带这些载体的微生物以及括号中的这些微生物在国际保存库(internationaldepositories)中的获取号。pAJ655 大肠杆菌AJ11882(FERM BP-136)
谷氨酸棒状杆菌SR8201(ATCC39135)pAJ1844大肠杆菌AJ11883(FERM BP-137)
谷氨酸棒状杆菌SR8202(ATCC39136)pAJ611 大肠杆菌AJ11884(FERM BP-138)pAJ3148谷氨酸棒状杆菌SR8203(ATCC39137)pAJ440 枯草芽孢杆菌AJ11901(FREM BP-140)pHC4 大肠杆菌AJ12617(FERM BP-3532)
为了通过将本发明的DNA同在棒状杆菌中具有功能的载体进行连接而制备重组DNA，以一种限制性酶消化该载体，该限制性酶提供一种对应于本发明的DNA的末端的末端。该连接一般通过使用诸如T4 DNA连接酶这样的连接酶而完成。
为将上述的重组DNA引入一种诸如棒状杆菌这样的宿主，任何迄今为止已被报道的已知转化方法均可被应用。例如，可以使用一种方法对受体细胞进行氯化钙处理从而提高其对DNA的通透性，该方法已经针对大肠杆菌K-12进行了报道(Mandel，M.and Higa，A.，J.Mol.Biol.，52，159(1970))，还可使用一种方法来从处于生长期的细胞中制备感受态细胞，随后将该DNA引入其中，该方法已经针对枯草芽孢杆菌进行了报道(Duncan，C.H.，Wilson，G.A.and Young，F.E.，Gene，1，153(1977))。除这些方法之外，还可使用一种方法来将DNA受体细胞制成原生质体或者原生质球，它们易于摄取重组DNA，随后将该重组DNA引入这些细胞，该方法已知可应用于枯草芽孢杆菌、放线菌和酵母(Chang，S.and Choen，S.N.，Molec.Gen.Genet.，168，111(1979)；Bibb，M.J.，Ward，J.M.and Hopwood，O.A.，Nature，274，398(1978)；Hinnen，A.，Hicks，J.B.and Fink，G.R.，Proc.Natl.Sci.USA，75，1929(1978))。棒状杆菌的转化可通过电脉冲方法有效地进行(参考Japanese Patent Laid-openNo.2-207791)。
关于诸如Corynebacterium thermoamonogenes这样的嗜热棒状杆菌的转化，通过用一种可以改变宿主细胞的细胞壁的结构的试剂对细胞进行处理，并且对一种含有DNA和这些细胞壁已经改变的细胞的溶液应用电脉冲来有效地完成。上述的试剂可以改变细胞壁的结构，使得在对一种含有经该试剂处理的细胞以及DNA的溶液进行电脉冲时，这些细胞能够摄入DNA(在下文中亦称为“细胞壁处理试剂”)。这样的试剂的例子包括抑制细菌细胞壁的正常合成的试剂，还包括对细菌细胞壁进行裂解的试剂。它们的具体例子包括溶菌酶、青霉素G、甘氨酸等。
这些细胞壁处理试剂均可单独使用，或者其中两种或者更多种可联合使用。上述试剂中溶菌酶和青霉素G较为优选，并且溶菌酶特别地优选。
进一步，Corynebacterium thermoamonogenes的转化还可通过对含有DNA和经物理方法进行了细胞壁弱化的细胞进行电脉冲来完成，这样的物理方法的例子如超声(FEMS Microbiology Letters，151，135-138(1987))。
为有效地表达本发明的DNA中所含有的基因，可在该宿主细胞中发挥功能的启动子如lac、trp和PL可以连接于该基因编码区域的上游。如果使用含有一种启动子的载体作为该载体，各基因、载体和启动子的连接可一步完成。
可以如上所述而生产的本发明的蛋白质，可根据需要从细胞抽提物或者培养基中纯化，该纯化通过常规的方法进行，例如，离子交换层析、凝胶过滤层析、吸附层析、盐析和溶解沉淀。
预期本发明的蛋白在热稳定性上十分优秀或者同谷氨酸棒状杆菌等的相应蛋白相比较在高温下表现出更高的活性。例如，乳发酵短杆菌的GDH在37℃附近表现出最高的活性，并且该活性在42℃附近明显地降低。但是，本发明的GDH在42℃下所表现出的GDH活性等于或者高于37℃下的活性。在一个优选的实施方案中，本发明的GDH在42℃附近表现出最大活性并且甚至在45℃下还表现出活性。
GDH活性的测量可通过，例如，将该酶加入至100mM Tris-HCl(pH8.0)、20mM NH4Cl、10mM α-酮戊二酸、0.25mM NADPH并且测定在340nm下的吸收的变化来进行(Molecular Microbiology 6，317-326(1992))。
进一步，乳发酵短杆菌的CS在23℃附近表现出最高的CS活性，并且该活性在33℃附近明显降低。与之相反，本发明的CS在37℃下所表现出的CS活性等于或者高于23℃下的活性。在一个优选的实施方案中，本发明的CS表现出随反应温度提高的活性直至约37℃，并且甚至在40℃下仍表现出同37℃下的活性相比约40％的活性。
CS活性的测量可通过，例如，在Enzymol.，13，3-11(1969)中所描述的方法来进行。
进一步，本发明的其它蛋白典型的具有以下的特征。异柠檬酸裂合酶在50℃下处理5分钟后具有30％或者更多的残余活性。磷酸果糖激酶在60℃下的活性等于或者高于在30℃下的活性。磷酸肌醇丙酮酸羧化酶在45℃下处理5分钟后具有50％或者更高的残余活性。顺乌头酸酶在50℃下处理3分钟后具有30％或者更高的残余活性。异柠檬酸脱氢酶在45℃下处理10分钟后具有50％或者更高的残余活性。2-酮戊二酸脱氢酶在50℃下处理10分钟后具有30％或者更高的残余活性。
本发明的蛋白质还可从诸如Corynebacteriumthermoaminogenes AJ12310菌株这样的Corynebacteriumthermoaminogenes的细胞抽提物中，以各自活性作为指数使用纯化酶的常规方法来获得，例如，离子交换层析、凝胶过滤层析、吸附层析、盐析和溶解沉淀。
在本发明的DNA中，pfk、pdhA、pc、ppc、acn、icd、gdh和gltA(这些DNA所编码的酶的名称见表1)可被引入L-氨基酸生产菌中，例如棒状杆菌，以增强它们的L-氨基酸生产能力。引入了本发明的DNA的棒状杆菌还被预期能够在高于通常温度下生产L-氨基酸。这些L-氨基酸包括L-谷氨酸、L-天冬氨酸、L-赖氨酸、L-精氨酸、L-脯氨酸、L-谷氨酰胺等等。
例如，预期引入了gdh基因或者gltA基因的L-谷氨酸生产菌，例如棒状杆菌能够在高于通常温度下生产L-谷氨酸。进一步，尽管乳发酵短杆菌的CS可能在通常的培养温度下，例如在31.5℃下不能发挥全部功能，但是通过引入本发明的gltA基因该活性可被增强。
进一步，dtsR1和dtsR2是编码赋予棒状杆菌表面活性剂抗性的蛋白质(DTSR蛋白)的基因，这些基因受到破坏的产L-谷氨酸棒状杆菌会产生大量的L-谷氨酸，甚至在存在的生物素的量足以使野生型菌株基本不能产生L-谷氨酸的情况下仍是如此。进一步，如果具有L-赖氨酸生产能力的产L-谷氨酸棒状杆菌的dtsR1和dtsR2基因被扩增，则该细菌被赋予一种生产显著数量的L-赖氨酸的能力(WO95/23224，Japanese Patent Laid-open(Kokai)No.10-234371/1998)。
srcB基因可被用于改善棒状杆菌以用于生产L-氨基酸，该生产通过在含有蔗糖的培养基中使用棒状杆菌来进行。
通过缺失产L-谷氨酸棒状杆菌等的aceA、accBC、lpd或odhA，它们的L-谷氨酸产量可获得提高。进一步，gluABCD为L-谷氨酸摄取系统的一种基因簇，并且通过缺失产L-谷氨酸棒状杆菌的gluA、gluB、gluC和gluD的其中一种到四种，积累于培养基中的L-谷氨酸的量可获得提高。本发明的aceA、accBC、lpd、odhA和gluABCD可被用于在染色体上破坏这些基因。
用于通过利用引入了本发明DNA的微生物来生产L-氨基酸的培养基可为通常的培养基，该培养基含有碳源、氮源、无机离子和其它所需的有机微量营养物。作为碳源，可以使糖类，例如葡萄糖、乳糖、半乳糖、果糖、蔗糖、blackstrap糖蜜以及淀粉水解物；醇类，例如乙醇和肌醇；或者有机酸，例如乙酸、延胡索酸、柠檬酸和琥珀酸。
作为氮源，可以使用无机铵盐，例如硫酸铵、柠檬酸铵、氯化铵、磷酸铵和乙酸铵、氨，有机氮源，例如蛋白胨、肉类抽提物、酵母抽提物、玉米浆和大豆水解物、氨气、氨水等等。
作为无机离子(或无机离子源)，所添加的为少量的磷酸钾、硫酸镁、铁离子、锰离子等等。对于有机微量营养物，添加所需的物质较为理想，例如所需的适量维生素B1、酵母抽提物等等。
在优选的情况下，培养在通气的条件下进行，该通气通过16到72小时的振荡、搅拌等而进行通气。培养的温度被控制在30℃到47℃之间，并且培养期间pH被控制在5到9。对于培养温度，培养可在适于未引入本发明的DNA的微生物的培养温度或比之更高的温度下进行。为进行pH的调节可以使用无机或者有机酸或碱物质、氨水等等。
L-氨基酸从发酵肉汤中的收集可通过已知方法的综合使用而完成，例如利用离子交换树脂、沉淀、结晶等等基于L-氨基酸的类型的工艺。
图表的简要解释


图1所示为来自Corynebacterium thermoaminogenes AJ12310菌株和乳发酵短杆菌2256菌株的谷氨酸脱氢酶的活性随温度的变化。
图2所示为来自该AJ12310菌株和2256菌株的谷氨酸脱氢酶的热稳定性。
图3所示为来自该AJ12310菌株和2256菌株的柠檬酸合成酶的活性随温度的变化。
图4所示为来自该AJ12310菌株和2256菌株的柠檬酸合成酶的热稳定性。
图5所示为来自该AJ12310菌株和2256菌株的异柠檬酸裂合酶的活性随温度的变化。
图6所示为来自该AJ12310菌株和2256菌株的异柠檬酸裂合酶的热稳定性。
图7所示为来自该AJ12310菌株和2256菌株的磷酸果糖激酶的活性随温度的变化。
图8所示为来自该AJ12310菌株和2256菌株的磷酸果糖激酶的热稳定性。
图9所示为来自该AJ12310菌株和2256菌株的磷酸烯醇丙酮酸羧化酶的活性随温度的变化。
图10所示为来自该AJ12310菌株和2256菌株的磷酸烯醇丙酮酸羧化酶的热稳定性。
图11所示为来自该AJ12310菌株和2256菌株的顺乌头酸酶的活性随温度的变化。
图12所示为来自该AJ12310菌株和2256菌株的顺乌头酸酶的热稳定性。
图13所示为来自该AJ12310菌株和2256菌株的异柠檬酸脱氢酶的活性随温度的变化。
图14所示为来自该AJ12310菌株和2256菌株的异柠檬酸脱氢酶的热稳定性。
图15所示为来自该AJ12310菌株和2256菌株的2-酮戊二酸脱氢酶的热稳定性。
图16所示为携带scrB基因的质粒pSCR155的结构。
图17所示为携带pdhA基因的质粒pPDHA-2的结构。
图18所示为一种pdhA基因扩增菌株的L-谷氨酸的产量(a)37℃(b)44℃。
图19所示为携带icd基因的质粒pICD-4的结构。
图20所示为一种icd基因扩增菌株的L-谷氨酸的产量(a)37℃(b)44℃。
图21所示为质粒pHSG299YGDH和pYGDH的结构。
图22所示为质粒pHSG299YCS和pYCS的结构。
实施本发明的最佳模式
在下文中，本发明将参考随后的实施例进行进一步具体地说明。
实施例1
<1>Corynebacterium thermoaminogenes的质粒库的产生
在CM2B液体培养基(1g/dl的酵母抽提物(产自Difco)、1g/dl的多蛋白胨(产自Nippon Seiyaku)、0.5g/dl的NaCl、10μg/dl的生物素，pH7.0(以KOH进行调节))中37℃下对Corynebacteriumthermoaminogenes AJ12310菌株进行15个小时的培养，并且使用一种染色体DNA抽提试剂盒(细菌基因组DNA纯化试剂盒)(AdvancedGenetic Technologies生产)从10ml的培养基中制备其染色体DNA。所获得DNA以限制性酶Sau3AI进行部分消化，并且用于0.8％琼脂糖凝胶电泳以进行DNA的分离。随后，从胶中切出对应于约4到6kb的DNA片段的一条带，并且通过使用一种DNA凝胶抽提试剂盒(GIBCOBRL，ConcertTM快速凝胶抽提系统)获得该目的大小的DNA片段。
以BamHI对质粒pHSG399进行完全消化，并且通过使用碱性磷酸酶(CIAP；Takara Shuzo制造)对其末端进行去磷酸化。通过使用由Takara Shuzo生产的DNA连接试剂盒对这一载体和上述的染色体DNA进行连接，并以所获得的重组载体对大肠杆菌JM109进行转化。转化子的选择在含有30μg/ml氯霉素、0.04mg/ml的IPTG(异丙基-β-D-硫代半乳糖苷)以及0.04mg/ml的X-gal(5-溴-4-氯-3-吲哚-β-D-半乳糖苷)的LB琼脂培养基(含有1.5g/dl琼脂)上进行，获得了约4000个白色菌落。
<2>用于各基因的扩增的引物的设计
对用于通过PCR从以上获得的质粒文库中选择含有各靶基因的克隆的引物进行了设计。这些靶基因在上文提及。
这些引物基于棒状杆菌的已知基因序列进行设计，即，基于同其它微生物的对应基因进行比较时所观察到的在氨基酸水平上的保守区域的序列进行设计。考虑到棒状杆菌的密码子使用性，对每个基因设计了复合的引物组。
为检测所制备的引物的特征，使用这些引物以及使用Corynebacterium thermoaminogenes AJ12310菌株的染色体DNA作为模板进行PCR以扩增各基因片段。其结果，当使用表2到7中上排所示的引物，并在这些表中标识为“用于获得部分片段的PCR条件”的条件下进行PCR时，对于所有的基因均观察到了扩增片段。引物序列后的括号内的数字为该引物在序列表中的序列号。这些引物被用作为上述的筛选的引物。
表2
表3
表4
表5
表6
表7
<3>通过PCR对质粒文库进行筛选
通过PCR从质粒文库中选出含有一种靶基因的克隆。从各质粒文库中挑出60个菌落，并且在两个LB琼脂培养基平板上进行复制。以各平板的60个菌落进行组合，接种入含有4ml LB液体培养基的测试试管中并且培养15个小时。随后，分别通过使用Promega生产的质粒DNA抽提试剂盒获得质粒混合物。通过使用这一质粒混合物作为模板并且使用针对个靶基因而制备的筛选引物，在各表中的“用于筛选的PCR条件”下进行PCR以选出一种克隆，从该克隆中可获得一种DNA片段，该片段的大小与使用染色体DNA作为模板所扩增的DNA相同。
通过使用一种Perkin-Elmer生产的Big Dye dye终止体循环测序试剂盒(terminator cycles equencing kit)来测定所扩增的DNA片段的核苷酸序列，并且对其同已知基因信息的同源性进行研究以确定是否获得了该靶基因。
对于lpd，由于以<2>中生产的引物扩增不出任何DNA片段，所以基于所测定的核苷酸序列而制备了其它的筛选引物。
<4>通过菌落PCR进行的对携带靶基因的克隆的选择
通过使用一种平板而进行了菌落PCR以选择含有该基因片段的克隆，该平板为质粒混合物的来源，对此已经证实了靶基因的扩增。该菌落PCR以表2-7所示的条件进行。
质粒DNA选自一种被选定的转化子，并且对被插入DNA片段的核苷酸序列进行了测定。当该靶基因的全长并未被插入该被插入DNA片段中，并且上游区域、下游区域或两者均被缺失时，基于被测定的核苷酸序列而制备了引物，利用这些引物通过使用Takara LA PCR体外克隆试剂盒(Takara Shuzo)获得了含有全长的靶基因核苷酸序列的基因片段。随后测定了其核苷酸序列。
LA PCR克隆的纲要如下。产生两种引物，这两种引物各具有该被插入DNA片段的两个区域之一。以多种限制性酶对Corynebacteriumthermoaminogenes AJ12310菌株的染色体DNA进行消化，并且与一种对应于各限制性酶的盒式引物进行连接。通过使用这样的DNA作为模板，用一种对应于远离该缺失区域位置的引物(S1)和一种对应于所制备的引物中的盒式引物之外位置的引物(C1)进行PCR。随后，用一种对应于接近该缺失区域的位置的引物(S2)和一种对应于所制备的引物中的盒式引物之内的位置的引物(C2)进行PCR。通过这一途径而获得了一种含有该缺失区域的DNA片段。通过将所获得的该DNA片段同已经获得的DNA片段进行连接而获得了一种含有该靶基因的全长DNA片段。由于该盒的5’末端没有磷酸基团，在该DNA片段的3’末端和该盒的5’末端的连接位置可能形成一种缺刻。因此，在第一次PCR中从C1引物开始的的DNA的合成终止于这一连接位置，并且由此而不能进行非特异性的扩增。因此可以达到特异性的扩增。
表2-7所示为用于LA PCR克隆的引物和反应条件。在这些表中，以“(N’)”提到的引物为用于基因上游缺失部分的克隆的引物，以“(C’)”提到的引物为用于基因下游缺失部分的克隆的引物。根据LA PCR克隆试剂盒附带的指导进行两次PCR。在这些表中所提及的引物中，用于第一次反应的引物(S1)在上排中显示，用于第二次反应的引物(S2)在下排中显示。
如上所述所获得的含有各基因的DNA片段的核苷酸序列用与上文相同的手段进行测定。这些核苷酸序列和可由这些核苷酸序列编码的氨基酸序列见SEQ ID NOS1-34。以这些序列号所示的序列在此后提及的序列表解释中加以概述。
对于scrB，没有发现任何开放读码框架。由于Corynebacteriumthermoaminogenes AJ12310菌株不具有蔗糖酶活性并且不具有任何蔗糖同化特征，从具有蔗糖同化特征的Corynebacteriumthermoaminogenes AJ12340和AJ12309中以类似方式获得了scrB基因。其结果，从这两个菌株中均获得了具有开放读码框架的DNA片段。
实施例2gdh和gltA基因的获得
<1>Corynebacterium thermoaminogenes的GDH活性的研究
Corynebacterium thermoaminogenes的野生型菌株AJ12310细胞在CM-2B琼脂培养基上生长(1g/dl的酵母抽提物(产自Difco)、1g/dl的多蛋白胨(产自Nippon Seiyaku)、0.5g/dl的NaCl、10μg/dl的生物素，KOH调节至pH7.0)。将这些细胞接种于一个含有20ml烧瓶培养基的500-ml体积的烧瓶中并且在37℃下培养17个小时(直到剩余的糖达到约1g/dl)，该烧瓶培养基中含有以下组合物。
类似地，乳发酵短杆菌2256菌株(ATCC13869)的细胞在CM-2B琼脂培养基上31.5℃下培养17个小时。
葡萄糖 3g/dl
KH2PO4 0.1g/dl
MgSO4·H2O 0.04g/dl
FeSO4·7H2O1mg/dl
MnSO4·4H2O1mg/dl
维生素B1-HCl200μg/L
生物素 50μg/L
(NH4)2SO4 1.5g/dl
大豆蛋白水解溶液 48mg/dl
(Memeno(T-N))
CaCO3(Official regent) 5g/dl(分开灭菌)
pH8.0(以KOH调节)
以1000rpm对约1ml的上述培养基进行1分钟的离心以除去CaCO3，并且以200mM的K-磷酸缓冲液(pH6.9)对细胞进行洗涤并且悬浮于300μl的相同缓冲液中。对所获得的细胞悬浮物进行5分钟的超声以破碎细胞，在1000rpm下离心30分钟以获得粗级酶溶液的上清。
使用上述的粗级酶溶液检测GDH活性的最佳反应温度和热稳定性。GDH活性的测量通过将该粗级酶溶液加至一种反应混合物(100mMTris-HCl(pH8.0)，20mM的NH4Cl，10mM的α-酮戊二酸钠，0.25mMNADPH)中并测量340nm下的吸收变化而进行。该粗级酶溶液的蛋白质含量通过Bradford方法(Bio-Rad蛋白质分析试剂盒)使用牛血清白蛋白作为标准通过对595nm吸收的测量而进行定量。该吸收通过使用HITACHI U-2000(Hitachi生产)而进行。
图1所示为在不同的反应温度下测得的GDH活性。尽管ATCC13869菌株在37℃附近表现出最高的GDH特异活性并且该活性在42℃附近明显降低，但是AJ12310菌株在42℃附近表现出最高的特异活性并且它甚至在45℃还表现出活性。
随即对GDH的热稳定性进行了测试。在反应之前，在65℃下放置粗级酶溶液0到30分钟，并且随后在30℃下测量该酶活性。结果见图2。从这些结果清晰可见，尽管ATCC13865菌株的GDH被5分钟的热处理所失活，AJ12310菌株的GDH甚至在30分钟的热处理后依然维持。另外，AJ12310菌株的粗级酶溶液甚至在65℃下处理90分钟后在GDH活性方面基本不表现出变化(数据未出示)。
<2>Corynebacterium thermoaminogenes的GCS活性的检测
使用粗级酶溶液检测CS的最佳反应温度和热稳定性，该粗级酶溶液以同实施例1相同的方式从Corynebacterium thermoaminogenesAJ12310菌株和乳发酵短杆菌ATCC13869菌株的细胞中制备。CS活性的测量通过将各粗级酶溶液加至一种反应混合物(100mM Tris-HCl(pH8.0)，0.1mM的DTNB(5，5’-dithiobis-(2-nitrobenzoicacid))，200mM的L-谷氨酸钠，0.3mM脂酰辅酶A)中并测量412nm处的吸收变化而进行。
图3所示为在不同的反应温度下测得的CS活性。ATCC13869菌株在23℃附近表现出最高的CS比活性并且该活性在33℃附近明显降低。但是，AJ12310菌株直至37℃附近仍然表现出依赖于反应温度的高度活性，并且它甚至在40℃还表现出相对于37℃下约40％的活性。
随后对CS的热稳定性进行了检测。在反应之前，将该粗级酶溶液在35-55℃下放置5分钟并且随后在30℃下测量该酶的活性。结果见图4。ATCC13869菌株的CS可被35-40℃的热处理灭活，而AJ12310菌株的CS甚至在50℃下处理后仍然维持约40％的活性。
<3>Corynebacterium thermoaminogenes的ghd基因的获得
对已经报道的多种微生物的gdh基因的核苷酸序列进行了比较。选中了一个含有高度保守的核苷酸序列的区域，并且基于该区域的核苷酸序列制备了具有SEQ ID NOS77和78所示的核苷酸序列的引物。
通过使用从Corynebacterium thermoaminogenes AJ12310菌株中制备的染色体DNA作为模板以及上述的引物进行PCR，该染色体DNA使用细菌基因组DNA纯化试剂盒(Advanced Genetic Technologies制造)进行制备。基于所获得的DNA片段，通过使用Takara LA PCR体外克隆试剂盒进行了基因组步移以获得完整的gdh基因。结果见SEQID NO79。进一步，从这一核苷酸序列推导出的氨基酸序列见SEQ IDNO80。
以类似的方式获得乳发酵短杆菌ATCC13869菌株的gdh基因，并且测定其核苷酸序列。结果见SEQ ID NO81。由这一核苷酸序列所编码的氨基酸序列见SEQ ID NO82。
对如上所述测定的Corynebacterium thermoaminogenes AJ12310菌株和乳发酵短杆菌ATCC13869菌株gdh基因的核苷酸序列和GDH的氨基酸序列，以及已知的谷氨酸棒状杆菌(C.glutamicum)ATCC13032的gdh基因和GDH氨基酸序列(Molecular Microbiology 6，317-326(1992))的同源性进行了研究。结果见表8(核苷酸序列)和表9(氨基酸序列)。
表8不同的gdh基因的核苷酸序列的同源性
表9不同的GDH的氨基酸序列的同源性
<4>Corynebacterium thermoaminogenes的gltA基因的获得
对已经报道的多种微生物的gltA基因的核苷酸序列进行了比较。选中了一个含有高度保守的核苷酸序列的区域并且基于该区域的核苷酸序列制备了具有SEQ ID NOS83和84所示的核苷酸序列的引物。
通过使用从Corynebacterium thermoaminogenes AJ12310菌株(FERM BP-1542)中制备的染色体DNA作为模板以及上述的引物7和8进行PCR，该染色体DNA使用细菌基因组DNA纯化试剂盒(AdvancedGenetic Technologies制造)进行制备，并且对约0.9kb的被扩增的核苷酸序列进行了测定。
根据所获得的谷氨酸棒状杆菌的gltA基因的核苷酸序列(Microbiol.，140，1817-1828(1994))而制备了SEQ ID NOS85、86、87和88的引物。以类似于上文的方式使用AJ12310的染色体DNA作为模板并且使用SEQ ID NOS85、86、87和88的引物进行PCR，对被扩增的DNA片段的核苷酸序列进行鉴定以测定该gltA基因的完整核苷酸序列。结果见SEQ ID NO89。进一步，由这一核苷酸序列而预期的氨基酸序列见SEQ ID NO90。
以类似的方式获得乳发酵短杆菌2256菌株的gltA基因并且测定其核苷酸序列。结果见SEQ ID NO91。这一核苷酸序列所编码的氨基酸序列见SEQ ID NO92。
对如上所述测定的Corynebacterium thermoaminogenes AJ12310菌株和乳发酵短杆菌ATCC13869菌株gltA基因的核苷酸序列和CS的氨基酸序列，以及已知的谷氨酸棒状杆菌(C.glutamicum)ATCC13032的gltA基因和CS氨基酸序列(Microbiol.，140，1817-1828(1994))的同源性进行了研究。结果见表10(核苷酸序列)和表11(氨基酸序列)。
表10不同的gltA基因的核苷酸序列的同源性
表11不同的CS的氨基酸序列的同源性实施例3Corynebacterium thermoaminogenes的scrB基因的获得
由于scrB基因片段如实施例1所示从Corynebacteriumthermoaminogenes AJ12309菌株的中获得，对基因的全序列的获得进行了尝试。首先，使用在SEQ ID NO45和SEQ ID NO46中所示的引物，以与实施例1相同的方式获得了一个部分片段。这些引物基于乳发酵短杆菌2256菌株的scrB序列(Japanese Patent Laid-openNo.08-196280/1996)进行合成。
另外，通过使用细菌基因组DNA纯化试剂盒(Advanced GeneticTechnologies Corp.)从AJ12309菌株中制备了染色体DNA。向0.5μg的该染色体DNA、50pmol的各种上述引物、4μl的dNTP混合物(每种2.5mM)、5μl的10×Z-Taq缓冲液(Taraka Shuzo)以及2U的Z-Taq(Taraka Shuzo)中加入无菌水以制备总体积为50μl的PCR反应缓冲液。使用上述的反应混合物以及热循环仪GeneAmp PCR系统9600(PE)进行PCR以扩增出约600bp的scrB部分片段，该PCR所用的循环为98℃下进行5秒钟的变性，在50℃下进行10秒钟的结合并且在72℃下进行20秒钟的延伸反应，该过程重复循环30次。
随后，通过使用LA PCR体外克隆试剂盒(Takara Shuzo)来测定scrB的全序列。所有这些过程均根据该LA PCR体外克隆试剂盒所附带的方案而进行。根据所获得的部分序列而合成了在SEQ ID NOS97、98、99和100中出示的序列。为进行用于对上游区域进行测序的第一个PCR反应，而使用了SEQ ID NOS95和97所示的引物以及作为模板的AJ12309菌株染色体DNA，该染色体DNA经EcoT14I消化。为进行第二个PCR反应而使用了SEQ ID NOS96和98所示的的引物。为进行用于对下游区域进行测序的第一个PCR反应而使用了SEQ IDNOS95和99所示的引物以及作为模板的AJ12309菌株染色体DNA，该染色体DNA经SalI(Takara Shuzo)消化。为进行第二个PCR反应而使用了SEQ ID NOS96和100所示的的引物。通过上述的步骤，对含有scrB的ORF的1656bp全长序列进行了测定。这一核苷酸序列示于SEQ ID NO93，所推定的氨基酸序列见SEQ ID NO94。实施例4异柠檬酸裂合酶、磷酸果糖激酶、磷酸烯醇丙酮酸羧化酶、顺乌头酸酶、异柠檬酸脱氢酶以及2-酮戊二酸脱氢酶的热稳定性的检测
对下列来自Corynebacterium thermoaminogenes的酶的热稳定性进行了检测。在本实施例中，蛋白质浓度使用牛血清白蛋白作为标准蛋白通过Bradford方法(所使用的为Bio-Rad蛋白质分析试剂盒)进行测量。进一步，除非另外说明，吸光度的测量通过使用HITACHIU-2000(Hitachi)来进行。
<1>异柠檬酸裂合酶
对来自Corynebacterium thermoaminogenes AJ12310菌株的异柠檬酸裂合酶(此后亦称为“ICL”)以及乳发酵短杆菌2256(ATCC13869)菌株的ICL的活性的热稳定性进行了测定。为进行活性测量，所使用的细胞在含有表2中提及的组合物的培养基中的培养，在所有碳源完全耗尽之前终止。该活性测量的方法在Dieter J.Reinscheid et al.，J.Bacteriol.，176(12)，3474(1994)中有所描述。具体地，以50mM的Tris缓冲液(pH7.3)洗涤这些细胞，在相同的缓冲液中进行悬浮，并且通过超声进行破碎(所使用的为KUBOTA制造的INSONATOR 201M，200W，5分钟)。超声之后，对悬浮物进行离心(13000×g，30分钟)来除去未破碎的细胞以制备粗级酶溶液。
将该粗级酶溶液加至一个反应系统，并且通过Hitachi分光光度计U-3210来测定不同温度下(30、40、50、60或70℃)的340nm处的吸收，该反应系统含有50mM MOPS-NaOH(pH7.3)、5mM二硫苏糖醇、15mM MgCl2、1mM EDTA、5mM D-苏型-异柠檬酸、0.2mM NADH以及18U的LDH(乳酸脱氢酶)。不同反应温度测量结果见图5。进一步，在50℃下对粗级酶溶液进行预处理(预处理时间5分钟或者15分钟)并且在37℃下对其活性进行测量。结果见图6。
其结果，AJ12310菌株的ICL在60℃下表现出最大活性，而2256菌株的ICL在约50℃下表现出最大活性。进一步，经5分钟的预处理后2256菌株的ICL完全失活，而AJ12310菌株的ICL经预处理5分钟之后保留了一半的活力。由此而证实了AJ12310菌株的ICL在高温下的稳定性。
表12用于ICL活性测量的培养基的组成
<2>磷酸果糖激酶
对来自Corynebacterium thermoaminogenes AJ12310菌株的磷酸果糖激酶(此后亦称为“PKF”)以及乳发酵短杆菌2256菌株的PKF的活性的热稳定性进行了测定。为进行活性测量，所使用的细胞在含有表13中提及的组合物的培养基中的培养，在所有的糖完全耗尽之前终止。该活性测量的方法在Michiko Mori et al.，Agric.Biol.Chem.，51(10)，2671(1994)中有所描述。具体地，以0.1M的Tris缓冲液(pH7.5)洗涤这些细胞，在相同的缓冲液中进行悬浮，并且通过超声进行破碎(所使用的为KUBOTA制造的INSONATOR 201M，200W，5分钟)。超声之后，对悬浮物进行离心(13000×g，30分钟)来除去未破碎的细胞以制备粗级酶溶液。
将该粗级酶溶液加至一个反应系统，并且通过Hitachi分光光度计U-3210来测定不同温度下(30、40、50、60或70℃)的340nm处的吸收，该反应系统含有100mM Tris缓冲液(pH7.5)、0.2mM NADH、10mM MgCl2、2mM NH4Cl、10mM KCl、0.2mM磷酸烯醇式丙酮酸、6.4mM果糖-6-磷酸、1mM ATP以及4μg的LDH/PK(丙酮酸激酶)。不同反应温度下的测量结果见图7。进一步，在50℃下对该粗级酶溶液进行预处理(预处理时间1、3、5分钟或者10分钟)并且在37℃下对其活性进行测量。结果见图8。
其结果，AJ12310菌株的PKF在约50℃下表现出最大活性，而2256菌株的PKF在约30℃下表现出最大活性。由此而证实了AJ12310菌株的PKF活性的最佳温度处于高温区域。
表13用于PKF活性测量的培养基的组成
<3>磷酸烯醇式丙酮酸羧化酶
对来自Corynebacterium thermoaminogenes AJ12310菌株的磷酸烯醇式丙酮酸羧化酶(此后亦称为“PEPC”)以及乳发酵短杆菌2256菌株的PEPC的活性的热稳定性进行了测定。
将在CM2-B琼脂培养基上生长的AJ12310菌株细胞接种于一个含有20ml烧瓶培养基(8g/dl的葡萄糖、0.1g/dl的KH2PO4、0.04g/dl的MgSO4·H2O、1mg/dl的FeSO4·7H2O、5mg/dl的MnSO4·4H2O、3g/dl的(NH4)2SO4、48mg/dl的TN(大豆蛋白水解溶液)、200μg/L的维生素B1、300μg/L的生物素、50μl/l的GD-113(消泡剂)、5g/dl的CaCO3(official regent，分开灭菌)，pH8.0(以KOH调节))的500-ml体积的烧瓶中并且在37℃下培养。2256菌株细胞在CM-2B琼脂培养基上在31.5℃下进行类似地培养。
上述令细胞生长至对数培养期的培养肉汤在1000rpm下离心1分钟以除去CaCO3，并且以洗涤缓冲液(100mM Tris/HCl pH8.0，10mMDTT，20％甘油)洗涤这些细胞3次，超声破碎细胞并且以15krpm离心10分钟以除去细胞碎片。上清以60krpm进行1小时的进一步离心以获得上清的粗级酶溶液。
通过使用上述的粗级酶溶液对PEPC活性的最佳反应温度和热稳定性进行检测。PEPC活性的测量通过将粗级酶溶液加至一种反应混合物(100mM Tris/H2SO4(pH8.5)、5mM磷酸烯醇式丙酮酸、10mM KHCO3、0.1mM乙酰辅酶A、0.15mM的NADH、10mM MgSO4、10U的苹果酸脱氢酶、0.1mM DTT)并且在800μl反应体积中测量340nm处的吸收变化来进行。
图9所示为在不同反应温度下测定的PEPC活性。2256菌株的活性在40℃下明显地减少，而AJ12310菌株甚至在40℃下基本不表现出活性的降低。
随后对PEPC的热稳定性进行了检测。在反应之前将该粗级酶溶液在45℃下放置0-20分钟，并且随后在20℃下测量该酶活性。结果见
图10。从结果清晰可见，2256菌株的PEPC活性在经过10分钟的热处理后基本丧失，AJ12310菌株的PEPC甚至在热处理20分钟后仍然保持。
这些结果显示了AJ12310菌株的PEPC在高温下的稳定性。
<4>顺乌头酸酶
测定了来自Corynebacterium thermoaminogenes AJ12310菌株的顺乌头酸酶(此后亦称为“ACN”)以及乳发酵短杆菌2256菌株的ACN，以及其热稳定性。
将在CM2-B琼脂培养基上生长的AJ12310菌株细胞接种于一个含有20ml烧瓶培养基的500-ml体积的烧瓶中并且在37℃下培养，该烧瓶培养基含有的组分与<3>中提及的组分相同。2256菌株细胞在CM-2B琼脂培养基上在31.5℃下进行类似地培养。
上述令细胞生长至对数培养期的培养肉汤在1000rpm下离心1分钟以除去CaCO3，并且以50mM Tris/HCl pH8.0洗涤这些细胞3次，超声破碎细胞并且以15krpm离心10分钟以获得上清的粗级酶溶液。
通过使用上述的粗级酶溶液对ACN活性的最佳反应温度和热稳定性进行检测。ACN活性的测量通过将粗级酶溶液加至一种反应混合物(20mM Tris/HCl(pH7.5)、50mM NaCl、20mM异柠檬酸三钠)并且在800μl反应体积中测量240nm处的吸收变化来进行。
图11所示为在不同反应温度下测定的ACN活性。同2256菌株相比较，AJ12310菌株在高温之下表现出更高的活性。
随后对ACN的热稳定性进行了检测。在反应之前将该粗级酶溶液在50℃下放置0-15分钟，并且随后在30℃下测量该酶活性。结果见
图12。从结果清晰可见，同2256菌株相比较，AJ12310的ACN表现出较少的由于热处理所造成活性降低。
这些结果显示了AJ12310菌株的ACN在高温下的稳定性。
<5>异柠檬酸脱氢酶
对来自Corynebacterium thermoaminogenes AJ12310菌株的异柠檬酸脱氢酶(此后亦称为“ICDH”)以及乳发酵短杆菌2256菌株的ICDH的活性的热稳定性进行了测定。
将在CM2-B琼脂培养基上生长的AJ12310菌株细胞接种于一个含有20ml烧瓶培养基的500-ml体积的烧瓶中并且在37℃下培养，该烧瓶培养基含有的组分与<3>中提及的组分相同。2256菌株细胞在CM-2B琼脂培养基上31.5℃下进行类似地培养。
上述令细胞生长至对数培养期的培养肉汤在1000rpm下离心1分钟以除去CaCO3，并且以50mM Tris/HCl pH7.5洗涤这些细胞3次，超声破碎细胞并且以15krpm离心10分钟以获得上清的粗级酶溶液。
通过使用上述的粗级酶溶液对ICDH活性的最佳反应温度和热稳定性进行检测。ICDH活性的测量通过将粗级酶溶液加至一种反应混合物(35mM Tris/HCl、0.35mM EDTA(pH7.5)、1.5mM MnSO4、0.1mM NADP、1.3mM异柠檬酸三钠)并且在800μl反应体积中测量340nm处的吸收变化来进行。
图13所示为在不同反应温度下测定的ICDH活性。2256菌株的活性在70℃下明显地减少，而AJ12310菌株甚至在70℃下基本观察不到活性的降低。
随后对ICDH的热稳定性进行了检测。在反应之前将该粗级酶溶液在45℃下放置0-15分钟，并且随后在30℃下测量该酶活性。结果见
图14。从结果清晰可见，对于2256菌株，经过15分钟的热处理后仅仅观察到15％的残余活性，而对于AJ12310菌株可观察到60％的残余活性。
这些结果显示了AJ12310菌株的ICDH在高温下的稳定性。
<6>2-酮戊二酸脱氢酶
对来自Corynebacterium thermoaminogenes AJ12310菌株的2-酮戊二酸脱氢酶(此后亦称为“ODHC”)以及乳发酵短杆菌2256菌株的ODHC进行了测试并对其热稳定性进行了测定。
为进行活性测量，所使用的细胞在含有表14中提及的组合物的培养基中的培养，在所有糖完全耗尽之前终止。该活性测量的方法在Isamu Shiio et al.，Agric.Biol.Chem.，44(8)，1897(1987)中有所描述。具体地，以0.2％的氯化钾洗涤这些细胞，在100mMTES-NaOH(pH7.5)，30％的甘油溶液中进行悬浮，并且通过超声进行破碎(所使用的为KUBOTA制造的INSONATOR 201M，200W，5分钟)。超声之后，对悬浮物进行离心(13000×g，30分钟)来除去未破碎的细胞，并且使用同样缓冲液和Sephadex-G25进行凝胶过滤以制备粗级酶溶液。
将该粗级酶溶液加至一个反应系统，并且通过Hitachi分光光度计U-3210来测定不同温度下(30、40、50、60或70℃)的365nm下的吸收，该反应系统含有100mM TES-NaOH(pH7.7)、5mM MgCl2、0.2mM辅酶A、0.3mM脱羧辅酶、1mMα-酮戊二酸、3mM L-半胱氨酸以及1mM的乙酰吡啶-腺嘌呤二核苷酸。不同反应温度测量结果见图5。进一步，在50℃下对粗级酶溶液进行预处理(预处理时间5分钟或者15分钟)并且在37℃下对其活性进行测量。结果见
图15。
其结果，经10分钟的预处理后2256菌株的ODHC完全失活，而AJ12310菌株的ODHC活性基本保持恒定并不因预热时间不同而变化，并且由此而证实了该菌株的ODHC在高温下的稳定性。
表14用于ODHC活性测量的培养基的组成实施例5以srcB基因进行的基因转移所造成的蔗糖同化能力的获得
由于Corynebacterium thermoaminogenes AJ12310菌株不具有蔗糖酶活性以及蔗糖同化能力，因此对是否能够通过将AJ12309菌株的scrB基因转移至该菌株中而将蔗糖同化能力赋予该菌株进行了探索。
<1>携带来自Corynebacterium thermoaminogenes AJ12309菌株的scrB的质粒的产生
为获得一个scrB基因片段，根据SEQ ID NOS93的核苷酸序列合成了SEQ ID NOS101和102中所示的引物，这些引物的两个末端均同SmaI序列进行连接。将无菌水加至0.5μg 12309菌株的染色体DNA、上述寡核苷酸各50pmol、4μl的dNTP混合物(各2.5mM)、5μl的10×Pyrobest缓冲液(Takara Shuzo)以及2U的Pyrobest聚合酶(Taraka Shuzo)以制备成总体积50μl的反应混合物。使用上述的反应混合物以及热循环仪GeneAmp PCR系统9600(PE)进行PCR，扩增出约1.7kbp含有scrB ORF的片段，该PCR所用的循环为98℃下进行10秒钟的变性，在55℃下进行30秒钟的结合并且在72℃下进行2分钟的延伸反应，该过程重复循环30次。
随后，以SmaI(Takara Shuzo)消化上述的扩增片段，并且与含有一个去磷酸化的复制起点的质粒pSAC4进行连接以制备pSCR155，该复制起点可在棒状杆菌中发挥作用，该质粒已经经过SmaI的消化。
图16所示为pSCR155的结构。pSAC4按下列步骤产生。为使大肠杆菌载体pHSG399(Takara Shuzo)能够在棒状杆菌中自行复制，向其中引入了来自能够在棒状杆菌自行复制的质粒pHM1519(Miwa，k.et al.，Agric.Biol.Chem.，48(1984)2901-2903)的复制起点(JapanesePatent Laid-open No.5-7491/1993)。具体地，以限制性酶BamHI和KpnI消化pHM1519，并且通过使用Takara Shuzo所生产的平末端化试剂盒将所获得的含有复制起点的片段平末端化，并且通过使用一种SalI连接物(Takara Shuzo制造)将该片段插入pHSG399的SalI位点以获得pSAC4。
<2>携带scrB基因的质粒向AJ12310菌株的转移
向Corynebacterium thermoaminogenes AJ12309菌株中引入上文中生产的pSCR155和携带有来自乳发酵短杆菌的scrB基因的质粒pSSM30BS(Japanese Patent Laid-open No.08-196280/1996)。该转化根据以下程序进行。将细胞接种于含有20％的蔗糖的CM-2B培养基中，接种量应使该培养基的OD660为0.1，并且在37℃下振荡培养直至OD660成为0.3。向该培养基内加入浓度为100μg/ml的溶菌酶，并且这些细胞被进一步培养2小时。以20％的蔗糖洗涤这些细胞3次，悬浮于20％的蔗糖中，加入从大肠杆菌JM110中收集的质粒，充分混合并且对其使用电脉冲(18KV/cm，300毫秒)以引入这些DNA。在含有20％的蔗糖的CM-2B培养基中使这些细胞进行过夜恢复培养之后，在含有5μg/ml氯霉素的CM-2B培养基上对转化子进行选择。具体地，转化通过电脉冲方法进行(Japanese Patent Laid-open No.12-204236/2000)，转化子的选择在含有5μg/ml氯霉素的CM2B培养基上37℃下进行。其结果，未获得任何携带含有来自乳发酵短杆菌的scrB基因的质粒pSSM30BS的转化子，而只获得携带含有来自Corynebacterium thermoaminogenes的scrB基因的质粒pSCR155的转化子。这一菌株被命名为AJ12310/pSCR155。
<3>对使用蔗糖作为碳源进行的AJ12310/pSCR155菌株的培养的评估
将上文制备的AJ12310/pSCR155接种于一种含有表15所示的组分的培养基中，并且在37℃下振荡培养22小时。在培养之后测量吸收值(OD)和残余的糖(RS)。结果见表16。其结果，经过证实，AJ12310菌株不能同化蔗糖并且因此不能进行生长，而引入了scrB基因的菌株AJ12310/pSCR155菌株变得能够同化蔗糖。
表15培养基组成
表16蔗糖培养结果实施例6通过pdhA基因扩增的菌株进行的L-谷氨酸的生产
<1>携带pdhA的质粒pPDHA-2的结构
通过对质粒文库的筛选而获得了来自Corynebacteriumthermoaminogenes AJ12309菌株的pdhA基因。具体地，使用质粒文库混合物作为模板以实施例1的表4所示的条件进行PCR，并且选出了一个克隆p21A，从该克隆中扩增出了一个DNA片段，该片段的大小与以染色体DNA作为模板进行的PCR所获得DNA片段相同。对这一质粒的DNA序列进行测定以确认全长的pdhA包含于其中。
以XbaI和KpnI对p21A进行消化以切割成一个含有全长pdhA基因和启动子区域的4kb的DNA片段。将这一含有pdhA基因的DNA片段插入pHSG299(Takara Shuzo)的XbaI和KpnI位点。随后，以XbaI消化该质粒，并且将一个通过以XbaI消化pXK4而获得的片段插入其中以制备pPDHA-2。pPDHA-2的制备过程见
图17。一种DNA连接试剂盒Ver.2(Takara Shuzo)被用于连接反应，大肠杆菌JM109菌株(Takara Shuzo)被用作为遗传操作的宿主。上述的pXK4按如下过程产生。用限制性酶BamHI和KpnI对一种棒状杆菌和大肠杆菌的穿梭质粒pHK4(Japanese Patent Laid-open No.5-7491/1993)进行消化，以获得一种含有复制起点的DNA片段，并且通过使用一种DNA平末端化试剂盒(平末端化试剂盒由Takara Shuzo制造)对所获得的片段进行平末端化，将其同一种XbaI连接头(由Takara Shuzo制造)进行连接并且插入pHSG299的XbaI位点以获得质粒pKX4。
<2>携带pdhA基因的质粒向AJ12310菌株中的转移
将上文生产的质粒pPDHA-2引入Corynebacteriumthermoaminogenes AJ12309菌株以制备一种可进行pdhA基因扩增的菌株。该转化以同实施例5相同的方式进行，并且在含有25μg/ml的卡那霉素的CM-2B培养基上对转化子进行选择以获得AJ12310/pPDHA-2菌株。
<3>用pdhA扩增的菌株进行的L-谷氨酸的生产
AJ12310菌株和上文获得的pdhA基因扩增的菌株AJ12310/pPDHA-2菌株均在CN-2B培养基上生长，将它们接种于一个500-ml体积的烧瓶中并且在37℃下进行振荡培养直至葡萄糖完全耗尽，该烧瓶含有20ml用于育种培养烧瓶的培养基，该培养基见表17。以2ml该培养物肉汤接种于一个500ml体积烧瓶中并且在37℃和44℃下作为主要培养物进行培养，该烧瓶含有20ml用于主要培养烧瓶的培养基，该培养基见表17。持续进行主要培养直至葡萄糖完全耗尽。培养之后测量该培养基的OD以及L-谷氨酸的积累量以测定该基因扩增在细胞生长和谷氨酸生产中的作用。OD的测量通过使用HITACHIU-2000分光光度计(Hitachi)来进行，L-谷氨酸含量的测量通过使用AS-210谷氨酸分析仪(Asahi Chemical Industry)来进行。结果见
图18。
pdhA基因扩增的菌株，即AJ12310/pPDHA-2菌株，同Aj12310菌株相比较表现出L-谷氨酸积累增加并且表现出OD的增加，因此表明该pdhA基因对于L-谷氨酸的产生是有效的。
表17用于pdhA基因扩增的菌株的评测的培养基
实施例7用icd基因扩增的菌株进行的L-谷氨酸的生产
<1>携带来自Corynebacterium thermoaminogenes AJ12310菌株的icd的质粒pICD-4的结构
根据SEQ ID NO29所示的AJ12310菌株的icd基因序列合成了一些引物，这些引物见SEQ ID NO103和SEQ ID NO104。将一个BglII位点引入这两种引物的5’末端。另外，通过使用基因组DNA纯化试剂盒(Edge Biosystems)来制备Corynebacteriumthermoaminogenes AJ12310菌株的基因组DNA。将无菌水加至作为模板的该染色体DNA、上述引物各100pmol、8μl的dNTP混合物(各2.5mM)、10μl的10×Pyrobest缓冲液II(Takara Shuzo)以及2.5U的Pyrobest聚合酶(Taraka Shuzo)，制备成总体积100μl的反应混合物。使用上述的反应混合物以及热循环仪TP240(Taraka Shuzo)进行PCR以扩增出约3.3kb的含有icd基因及其启动子的DNA片段，该PCR所用的循环为98℃下进行10秒钟的变性，在55℃下进行1分钟的结合并且在72℃下进行4分钟的延伸反应，该过程重复循环30次。
随后，以BglII消化这一含有icd基因的DNA片段并且在BamHI位点同pHSG299(Taraka Shuzo)进行连接。随后以XbaI对这一质粒进行处理，并且将一种通过以XbaI消化pXK4而获得的片段插入该质粒中以结构pICD-4。pICD-4的结构程序见
图19。一种DNA连接试剂盒Ver.2(Takara Shuzo)被用于连接反应，大肠杆菌JM109菌株(Takara Shuzo)被用作为遗传操作的宿主。
<2>携带icd基因的质粒向AJ12310菌株中的转移
将上文所产生的质粒pICD-4引入Corynebacteriumthermoaminogenes AJ12310菌株以制备一种icd基因扩增的菌株。该转化以与实施例5相同的方式进行，并且在含有25μg/ml的卡那霉素的CM-2B培养基上对转化子进行选择以获得AJ12310/pICD-4菌株。
<3>通过icd扩增菌株进行的L-谷氨酸的生产
通过实施例6所描述的培养方法对AJ12310菌株和其icd扩增菌株AJ12310/pICD-4进行培养评估。结果见图20。icd扩增菌株AJ12310/pICD-4菌株同AJ12310菌株相比较，表现出L-谷氨酸积累增加以及OD的提高，由此表明icd基因的扩增对于L-谷氨酸的生产是有效的。
实施例8用gdh基因扩增菌株进行的L-谷氨酸的生产
<1>携带来自Corynebacterium thermoaminogenes AJ12310菌株的gdh的质粒结构
根据SEQ ID NO79所示的AJ12310菌株的gdh基因序列合成了一些引物，这些引物见SEQ ID NO105和SEQ ID NO106。另外，通过使用细菌基因组DNA纯化试剂盒(Advanced GeneticTechnologies Corp.)制备AJ12310菌株的基因组DNA。将无菌水加至0.5μg作为模板的该染色体DNA、上述引物各10pmol、8μl的dNTP混合物(各2.5mM)、5μl的10×LA Taq缓冲液(Takara Shuzo)以及2U的LA Taq(Taraka Shuzo)，制备成总体积50μl的反应混合物。使用上述的反应混合物以及热循环仪TP240(Taraka Shuzo)进行PCR以扩增出约2kb的含有gdh基因及其启动子的DNA片段，该PCR所用的循环为94℃下进行30秒钟的变性，在55℃下进行1秒种的结合并且在72℃下进行3分钟的延伸反应，该过程重复循环30次。以PstI(Takara Shuzo)消化所获得的扩增片段，将其与经过PstI完全消化的pHSG299(Takara Shuzo)混合并同其进行连接。Takara Shuzo生产的一种DNA连接试剂盒Ver.2被用于连接反应。连接之后，以连接产物对大肠杆菌JM109的感受态细胞(Takara Shuzo出产)进行转化，涂布于含有10μg/ml的IPTG(异丙基-β-D-硫代牛乳糖苷)、40μg/ml的X-Gal(5-溴-4-氯-3-吲哚-β-D-半乳糖苷)以及40μg/ml的氯霉素的L培养基平板上(10g/l的细菌蛋白胨、5g/l的细菌酵母抽提物、5g/l的NaCl、15g/l的琼脂，pH7.2)，并且进行过夜培养。挑选出现的白色菌落并且进行单菌落分离以获得转化子。
通过碱法从转化子中制备质粒(由日本生物科学和生物工程学会编辑的生物工程实验课本，p.105，Baifukan，1992)并且制作其限制性内切图谱。具有与图21所示的图谱等同的限制性图谱的质粒被命名为pHSG299YGDH。
将一种在棒状杆菌中发挥作用的复制起点引入pHSG299YGDH。具体地，以一种限制性酶XbaI消化pXC4以获得一种含有来自pHM1519的复制起点的片段，并且将该片段同经过XbaI完全消化的pHSG299YGDH进行混合并且同其进行连接。以上述途径制备质粒并且将具有与图21所示的图谱等同的限制性图谱的质粒命名为pYGDH。除了使用pHSG399(Cmr)代替pHSG299之外，pXC4的结构方式与实施例6中提及的pXK4的结构相同。
<2>携带gdh基因的质粒向AJ12310中的转移
将上文所产生的质粒引入Corynebacterium thermoaminogenesAJ12310菌株以制备一种可进行gdh基因扩增的菌株。该转化以与实施例5相同的方式进行，并且在31℃下在含有25μg/ml的卡那霉素的CM-2B培养基上对转化子进行选择以获得AJ12310/pYGDH菌株。
<3>通过gdh扩增菌株进行的的L-谷氨酸的生产
分别将在CM-2B培养基上生长的AJ12310菌株和上文获得的gdh扩增菌株AJ12310/pYGDH接种于500ml体积的烧瓶中并且在37℃下进行振荡培养直至葡萄糖被耗尽，该烧瓶中含有20ml用于育种培养的培养基，该培养基见表18。以2ml该培养物肉汤接种于一个500ml体积烧瓶中并且在37℃和44℃下作为主要培养物而培养，该烧瓶含有20ml用于主要培养烧瓶的培养基，该培养基见表19。持续进行主要培养直至葡萄糖完全耗尽。培养完成之后测量该培养基的OD620以及L-谷氨酸的积累量，以测定该基因扩增在细胞形成和谷氨酸生产中的作用。OD的测量通过使用HITACHIU-2000分光光度计(Hitachi)来进行，L-谷氨酸含量的测量通过使用AS-210谷氨酸分析仪(Asahi ChemicalIndustry)来进行。
表18用于育种培养的培养基的组成
表19用于主要培养的培养基的组成
培养的结果见表20和21。在37℃下，该gdh扩增菌株同亲本菌株AJ12310相比较表现出较高的糖类消耗速率、较为良好的生长以及获得较高的OD。进一步，L-谷氨酸积累和产率被显著地提高，即，在37％下提高5-7％。在44℃下产率也被提高，获得的OD也有所增加。在另一方面，在gdh扩增菌株中已经证实α-酮戊二酸的积累有所减少。这些结果表明gdh基因的扩增对于L-谷氨酸的产率的改善和副产物的减少是有效的。
表20可进行gdh扩增的菌株的培养结果(37℃)
表21 gdh扩增菌株的培养结果(44℃)实施例9通过gltA基因扩增的菌株进行的L-谷氨酸生产
<1>来自Corynebacterium thermoaminogenes的携带gltA基因的质粒的结构
根据SEQ ID NO89所示的AJ12310菌株的gltA基因序列合成了一些引物，这些引物见SEQ ID NO107和SEQ ID NO108。
另外，通过使用细菌基因组DNA纯化试剂盒(Advanced GeneticTechnologies Corp.)来制备AJ12310菌株的基因组DNA。将无菌水加至0.5μg作为模板的该染色体DNA、上述引物各10pmol、8μl的dNTP混合物(各2.5mM)、10μl的10×Pyrobest-Taq缓冲液(Takara Shuzo)以及2U的Pyrobest Taq(Taraka Shuzo)，制备成总体积100μl的反应混合物。使用上述的反应混合物以及热循环仪TP240(TarakaShuzo)进行PCR以扩增出约2kb的含有gltA基因及其启动子的DNA片段，该PCR所用的循环为94℃下进行30秒钟的变性，在45℃下进行30秒种的结合并且在72℃下进行3分钟的延伸反应，该过程重复循环30次。以KpnI(Takara Shuzo)消化所获得的扩增片段，将其与经过KpnI完全消化的pHSG299(Takara Shuzo)混合并同其进行连接。Takara Shuzo生产的一种DNA连接试剂盒Ver.2被用于连接反应。连接之后，以连接产物对大肠杆菌JM109的感受态细胞(Takara Shuzo出产)进行转化，涂布于含有10μg/ml的IPTG(异丙基-β-D-硫代半乳糖苷)、40μg/ml的X-Gal(5-溴-4-氯-3-吲哚-β-D-半乳糖苷)以及40μg/ml的氯霉素的L培养基平板上(10g/l的细菌蛋白胨、5g/l的细菌酵母抽提物、5g/l的NaCl、15g/l的琼脂，pH7.2)，并且进行过夜培养。挑选出现的白色菌落并且进行单菌落分离以获得转化子。
通过碱法从转化子中制备质粒(由日本生物科学和生物工程学会编辑的生物工程实验课本，p.105，Baifukan，1992)并且制作其限制性内切图谱。具有与图22所示的图谱等同的限制性图谱的质粒被命名为pHSG299YCS。
将一种在棒状杆菌中可以复制的复制起点引入pHSG299YCS。具体地，以一种限制性酶XbaI消化pXC4以获得一种含有来自pHM1519的复制起点的片段，并且将该片段同经过XbaI完全消化的pHSG299YCS进行混合并且同其进行连接。以上述途径制备质粒并且将具有与图22所示的图谱等同的限制性图谱的质粒命名为pYCS。
<2>携带gltA基因的质粒向AJ12310中的转移
将上文所产生的质粒引入Corynebacterium thermoaminogenesAJ12310菌株以制备一种gltA基因扩增的菌株。该转化以与实施例5相同的方式进行，并且在含有25μg/ml的卡那霉素的CM-2B培养基上对转化子进行选择以获得AJ12310/pYCS菌株。
<3>通过gltA扩增菌株进行的的L-谷氨酸的生产
分别将在CM-2B培养基上生长的AJ12310菌株和上文获得的gltA扩增菌株AJ12310/pYCS以与实施例8相同的方式进行培养。培养的结果见表22和表23。在培养温度37℃和44℃下，CS增强的菌株与其亲代菌株相比较表现出谷氨酸积累增加。进一步，该gltA扩增菌株表现出L-天冬氨酸和L-赖氨酸的积累的降低，它们合成自草酰乙酸。
这些结果表明gltA基因的扩增对于L-谷氨酸的产率的改善和副产物的减少是有效的。
表22可进行gltA扩增的菌株的培养结果(37℃)
表23可进行gltA扩增的菌株的培养结果(44℃)
SEQ ID NO1aceA，核苷酸序列
SEQ ID NO2aceA，氨基酸序列
SEQ ID NO3accBC，核苷酸序列
SEQ ID NO4accBC，氨基酸序列
SEQ ID NO5dtsR1，核苷酸序列
SEQ ID NO6dtsR1，氨基酸序列
SEQ ID NO7dtsR2，核苷酸序列
SEQ ID NO8dtsR2，氨基酸序列
SEQ ID NO9pfk，核苷酸序列
SEQ ID NO10pfk，氨基酸序列SEQ ID NO11scrB(AJ12340)，核苷酸序列SEQ ID NO12scrB(AJ12340)，氨基酸序列SEQ ID NO13scrB(AJ12309)，核苷酸序列SEQ ID NO14scrB(AJ12309)，氨基酸序列SEQ ID NO15scrB(AJ12310)，核苷酸序列SEQ ID NO16gluABCD，核苷酸序列SEQ ID NO17gluABCD，氨基酸序列SEQ ID NO18gluABCD，氨基酸序列SEQ ID NO19gluABCD，氨基酸序列SEQ ID NO20gluABCD，氨基酸序列SEQ ID NO21pdhA，核苷酸序列SEQ ID NO22pdhA，氨基酸序列SEQ ID NO23pc，核苷酸序列SEQ ID NO24pc，氨基酸序列SEQ ID NO25ppc，核苷酸序列SEQ ID NO26ppc，氨基酸序列SEQ ID NO27acn，核苷酸序列SEQ ID NO28acn，氨基酸序列SEQ ID NO29icd，核苷酸序列SEQ ID NO30icd，氨基酸序列SEQ ID NO31lpd，核苷酸序列SEQ ID NO32lpd，氨基酸序列SEQ ID NO33odhA，核苷酸序列SEQ ID NO34odhA，氨基酸序列SEQ ID NO79gdh(AJ12310)，核苷酸序列SEQ ID NO80gdh(AJ12310)，氨基酸序列SEQ ID NO81gdh(2256)，核苷酸序列SEQ ID NO82gdh(2256)，氨基酸序列SEQ ID NO89gltA(AJ12310)，核苷酸序列SEQ ID NO90gltA(AJ12310)，氨基酸序列SEQ ID NO91gltA(2256)，核苷酸序列SEQ ID NO92gltA(2256)，氨基酸序列
SEQ ID NO93scrB(AJ12309)，核苷酸序列
SEQ ID NO94scrB(AJ12309)，氨基酸序列
工业应用
根据本发明，编码来自Corynebacterium thermoaminogenes的氨基酸生物合成途径酶的基因或者编码氨基酸向细胞内摄入的蛋白质的基因。
本发明的基因可被用于上述酶或者蛋白质的生产或者氨基酸生产细菌的育种。
序列表<110>Ajinomoto Co.，Inc.<120>来自棒状嗜热细菌的氨基酸生物合成途径的酶的基团<130>B691SMOP1072<140><141>2000-10-04<150>JP 11-282716<151>1999-10-04<150>JP 11-311147<151>1999-11-01<150>JP 2000-120687<151>2000-04-21<160>108<170>PatentIn Ver.2.0<210>1<211>1980<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(577)..(1869)<400>1tgcattccac cgacggtcac gcgttcggtc ttgtcagcgg cgtcaatctg ctgatggttc 60atgcaaagct ccttcgaagc aagagatcgg gtgtgtgcgg gcacctatcg ggggaagccc 120tcgctgcgcc ccagggggag ctggcgatgt gaccaggtta agtgataacc atcaccttgc 180caatgggttt gcgaacttta ccgtgacgct acccccgctt ttgtttgatc acaccagctc 240gaaggctgtc gcttttccga agatgcacgt gaagtggcaa atccttgcca cccgaggttt 300tcccagtaca aacgtactag tgatgaggat cacggggaac attgtggaga ttgcactttg 360caatatttgc aaaaggggtg actacccccg cgcaaaactt aaaaacccaa atccgttgac 420ggacccatgc ccgatgaagc aatgtgtgaa gcacgccacc ggaacacagg ttgtggatca 480ctcaccatga tgtgggggat tcgcatcaca cagtgtgcag ggcggcacct ctaccgaatg 540cgccttacag cagcaccaag aagaagtgac tcttag atg tca aac gtt gga acg 594
Met Ser Asn Val Gly Thr
1 5cca cgt acc gca cag gaa atc cag cag gat tgg gac acc aac cca cgc 642Pro Arg Thr Ala Gln Glu Ile Gln Gln Asp Trp Asp Thr Asn Pro Arg
10 15 20tgg aac gga atc acc cgc gac tac acc gct gag cag gta gct gag ctc 690Trp Asn Gly Ile Thr Arg Asp Tyr Thr Ala Glu Gln Val Ala Glu Leu
25 30 35cag ggc agc gtc gtc gag gag cac acc ctc gca aag cgc ggc gcc gag 738Gln Gly Ser Val Val Glu Glu His Thr Leu Ala Lys Arg Gly Ala Glu
40 45 50atc ctg tgg gat gca gtt tcc gca gag ggc gac gac tac atc aac gca 786Ile Leu Trp Asp Ala Val Ser Ala Glu Gly Asp Asp Tyr Ile Asn Ala 55 60 65 70ctg ggc gcc ctt acc ggt aac cag gct gtc cag cag gtc cgt gcc ggc 834Leu Gly Ala Leu Thr Gly Asn Gln Ala Val Gln Gln Val Arg Ala Gly
75 80 85ctg aag gct gtc tac ctc tcc ggc tgg cag gtc gca ggt gac gcc aac 882Leu Lys Ala Val Tyr Leu Ser Gly Trp Gln Val Ala Gly Asp Ala Asn
90 95 100ctc gcc ggt cac acc tac ccc gac cag tcc ctg tac ccg gcg aac tcc 930Leu Ala Gly His Thr Tyr Pro Asp Gln Ser Leu Tyr Pro Ala Asn Ser
105 110 115gtc ccg aac gtt gtc cgt cgc atc aac aac gca ctg ctg cgc gcc gat 978Val Pro Asn Val Val Arg Arg Ile Asn Asn Ala Leu Leu Arg Ala Asp
120 125 130gag atc gca cgc gtc gag ggt gac acc tcc gtc gac aac tgg ctc gtc 1026Glu Ile Ala Arg Val Glu Gly Asp Thr Ser Val Asp Asn Trp Leu Val135 140 145 150ccg atc gtc gcc gac ggc gag gcc ggc ttc ggt ggc gcc ctc aac gtc 1074Pro Ile Val Ala Asp Gly Glu Ala Gly Phe Gly Gly Ala Leu Asn Val
155 160 165tac gag ctc cag aag ggc atg atc acc gct ggt gcc gca ggc acc cac 1122Tyr Glu Leu Gln Lys Gly Met Ile Thr Ala Gly Ala Ala Gly Thr His
170 175 180tgg gag gat cag ctc gct tcc gag aag aag tgt ggc cac ctc ggt ggc 1170Trp Glu Asp Gln Leu Ala Ser Glu Lys Lys Cys Gly His Leu Gly Gly
185 190 195aag gtc ctc atc ccg acc cag cag cac atc cgc acc ctg aac tcc gcc 1218Lys Val Leu Ile Pro Thr Gln Gln His Ile Arg Thr Leu Asn Ser Ala
200 205 210cgc ctg gca gct gac gtg gcc aac acc ccg acc gtc gtc atc gcc cgc 1266Arg Leu Ala Ala Asp Val Ala Asn Thr Pro Thr Val Val Ile Ala Arg215 220 225 230acc gac gca gag gcc gcc acc ctg atc acc tct gat gtt gat gag cgc 1314Thr Asp Ala Glu Ala Ala Thr Leu Ile Thr Ser Asp Val Asp Glu Arg
235 240 245gac cgc cca ttc atc acc ggc gag cgc acc gcc gag ggc tac tac cac 1362Asp Arg Pro Phe Ile Thr Gly Glu Arg Thr Ala Glu Gly Tyr Tyr His
250 255 260gtc aag ccg ggt ctc gag ccc tgc atc gca cgt gcg aag tcc tac gct 1410Val Lys Pro Gly Leu Glu Pro Cys Ile Ala Arg Ala Lys Ser Tyr Ala
265 270 275ccc tac gca gac atg atc tgg atg gag acc ggc acc cct gac ctc gag 1458Pro Tyr Ala Asp Met Ile Trp Met Glu Thr Gly Thr Pro Asp Leu Glu
280 285 290ctg gcc aag aag ttc gcc gag ggc gtc cgc agc gag ttc ccg gac cag 1506Leu Ala Lys Lys Phe Ala Glu Gly Val Arg Ser Glu Phe Pro Asp Gln295 300 305 310ctg ctg tcc tac aac tgc tcc ccg tcc ttc aac tgg tct gca cac ctc 1554Leu Leu Ser Tyr Asn Cys Ser Pro Ser Phe Asn Trp Ser Ala His Leu
315 320 325gag gcc gac gag atc gct aag ttc cag aag gaa ctg ggt gcc atg ggc 1602Glu Ala Asp Glu Ile Ala Lys Phe Gln Lys Glu Leu Gly Ala Met Gly
330 335 340ttc aag ttc cag ttc atc acc ctg gct ggc ttc cac tcc ctc aac tac 1650Phe Lys Phe Gln Phe Ile Thr Leu Ala Gly Phe His Ser Leu Asn Tyr
345 350 355ggt atg ttc gac ctg gct tac ggc tac gcc cgt gaa ggc atg ccc gcc 1698Gly Met Phe Asp Leu Ala Tyr Gly Tyr Ala Arg Glu Gly Met Pro Ala
360 365 370ttc gtc gac ctg cag aac cgt gag ttc aag gca gct gag gag cgc ggc 1746Phe Val Asp Leu Gln Asn Arg Glu Phe Lys Ala Ala Glu Glu Arg Gly375 380 385 390ttc acc gcc gtc aag cac cag cgt gag gtc ggc gcc ggc tac ttc gac 1794Phe Thr Ala Val Lys His Gln Arg Glu Val Gly Ala Gly Tyr Phe Asp
395 400 405acc atc gcc acc acc gtt gac ccg aac tcc tcc acc acc gcg ctg aag 1842Thr Ile Ala Thr Thr Val Asp Pro Asn Ser Ser Thr Thr Ala Leu Lys
410 415 420ggt tcc acc gag gaa tgc cag ttc cac taggaaccac ctgatgcggt 1889Gly Ser Thr Glu Glu Cys Gln Phe His
425 430gccgtatggc ctgacggcac cgcccctccc tttgcactcc agtactcctt tgtgcacatc 1949ggccatctcc acaccgcgcg ccccgccacc t1980<210>2<211>431<212>PRT<213>Corynebacterium thermoaminogenes<400>2Met Ser Asn Val Gly Thr Pro Arg Thr Ala Gln Glu Ile Gln Gln Asp 1 5 10 15Trp Asp Thr Asn Pro Arg Trp Asn Gly Ile Thr Arg Asp Tyr Thr Ala
20 25 30Glu Gln Val Ala Glu Leu Gln Gly Ser Val Val Glu Glu His Thr Leu
35 40 45Ala Lys Arg Gly Ala Glu Ile Leu Trp Asp Ala Val Ser Ala Glu Gly
50 55 60Asp Asp Tyr Ile Asn Ala Leu Gly Ala Leu Thr Gly Asn Gln Ala Val 65 70 75 80Gln Gln Val Arg Ala Gly Leu Lys Ala Val Tyr Leu Ser Gly Trp Gln
85 90 95Val Ala Gly Asp Ala Asn Leu Ala Gly His Thr Tyr Pro Asp Gln Ser
100 105 110Leu Tyr Pro Ala Asn Ser Val Pro Asn Val Val Arg Arg Ile Asn Asn
115 120 125Ala Leu Leu Arg Ala Asp Glu Ile Ala Arg Val Glu Gly Asp Thr Ser
130 13 140Val Asp Asn Trp Leu Val Pro Ile Val Ala Asp Gly Glu Ala Gly Phe145 150 155 160Gly Gly Ala Leu Asn Val Tyr Glu Leu Gln Lys Gly Met Ile Thr Ala
165 170 175Gly Ala Ala Gly Thr His Trp Glu Asp Gln Leu Ala Ser Glu Lys Lys
180 185 190Cys Gly His Leu Gly Gly Lys Val Leu Ile Pro Thr Gln Gln His Ile
195 200 205Arg Thr Leu Asn Ser Ala Arg Leu Ala Ala Asp Val Ala Asn Thr Pro
210 215 220Thr Val Val Ile Ala Arg Thr Asp Ala Glu Ala Ala Thr Leu Ile Thr225 230 235 240Ser Asp Val Asp Glu Arg Asp Arg Pro Phe Ile Thr Gly Glu Arg Thr
245 250 255Ala Glu Gly Tyr Tyr His Val Lys Pro Gly Leu Glu Pro Cys Ile Ala
260 265 270Arg Ala Lys Ser Tyr Ala Pro Tyr Ala Asp Met Ile Trp Met Glu Thr
275 280 285Gly Thr Pro Asp Leu Glu Leu Ala Lys Lys Phe Ala Glu Gly Val Arg
290 295 300Ser Glu Phe Pro Asp Gln Leu Leu Ser Tyr Asn Cys Ser Pro Ser Phe305 310 315 320Asn Trp Ser Ala His Leu Glu Ala Asp Glu Ile Ala Lys Phe Gln Lys
325 330 335Glu Leu Gly Ala Met Gly Phe Lys Phe Gln Phe Ile Thr Leu Ala Gly
340 345 350Phe His Ser Leu Asn Tyr Gly Met Phe Asp Leu Ala Tyr Gly Tyr Ala
355 360 365Arg Glu Gly Met Pro Ala Phe Val Asp Leu Gln Asn Arg Glu Phe Lys
370 375 380Ala Ala Glu Glu Arg Gly Phe Thr Ala Val Lys His Gln Arg Glu Val385 390 395 400Gly Ala Gly Tyr Phe Asp Thr Ile Ala Thr Thr Val Asp Pro Asn Ser
405 410 415Ser Thr Thr Ala Leu Lys Gly Ser Thr Glu Glu Cys Gln Phe His
420 425 430<210>3<211>2381<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(577)..(2349)<400>3agcaggccgt gttgccgaac ggcaacttcc gcagccgcaa ggagatcgag gaggtgtact 60cgcacctcaa ccctgccgag gacaccgtgg tgtactgccg cgtgggtgac cgcgcggccc 120acacctggtt cgtgttgaag tacctgctgg ggtttgaaaa cgtccgcaac tatgacggtt 180cctggtccga gtggggcaac atggtgcgca tgcccatcgt ccagggtgat gagccgggct 240cactctagtc accccggggt cacctccctg gtcacccccg taccctcccg ggtacacccc 300ggggacgggg tgtgacctgg atctcccctg catgtggaca ccgggaaact ttgcctggga 360aatgaccatc cagtaccgta atgcgggtat gttaacgcgg tcacagggta caccagaatc 420cggatcgtct aaccccctta gcgggattcg ctaaaagatc accgagttag tgtgcaagaa 480taatgctgat cgcaggggca ctgtcatacg ctgtcatgca gtcaatgaac agtgcggtgc 540tctgtcgtga agaaaatcaa aaccaggagg gtttta gtg tca gtc gag acc agg 594
Val Ser Val Glu Thr Arg
1 5aag atc acc aag gta ctt gtc gcc aac cgt ggt gaa atc gca atc cgt 642Lys Ile Thr Lys Val Leu Val Ala Asn Arg Gly Glu Ile Ala Ile Arg
10 15 20gtt ttc cgc gca gca cgg gat gaa ggc atc gcc tct gtc gcc gtc tac 690Val Phe Arg Ala Ala Arg Asp Glu Gly Ile Ala Ser Val Ala Val Tyr
25 30 35gcg gag ccg gac gca gat gcc cct ttc gtc gag tat gcc gat gag gcc 738Ala Glu Pro Asp Ala Asp Ala Pro Phe Val Glu Tyr Ala Asp Glu Ala
40 45 50ttc gca ctc ggt ggc cag act tcc gca gag tcc tac ctc gtc att gac 786Phe Ala Leu Gly Gly Gln Thr Ser Ala Glu Ser Tyr Leu Val Ile Asp 55 60 65 70aag atc att gac gca gca cgc aag tcc ggt gca gac gct gtc cac ccc 834Lys Ile Ile Asp Ala Ala Arg Lys Ser Gly Ala Asp Ala Val His Pro
75 80 85ggc tac ggc ttc ctc gcc gag aac gcc gat ttc gct gaa gct gtc atc 882Gly Tyr Gly Phe Leu Ala Glu Asn Ala Asp Phe Ala Glu Ala Val Ile
90 95 100aac gag ggc ctg atc tgg atc gga cca tcc cct gag tcc atc cgt tcc 930Asn Glu Gly Leu Ile Trp Ile Gly Pro Ser Pro Glu Ser Ile Arg Ser
105 110 115ctc ggt gac aag gtc acc gca cgc cac atc gcc aac aac gcc aac gca 978Leu Gly Asp Lys Val Thr Ala Arg His Ile Ala Asn Asn Ala Asn Ala
120 125 130ccg atg gca ccg ggc acc aag gag cct gtc aag gac gcc gct gag gtt 1026Pro Met Ala Pro Gly Thr Lys Glu Pro Val Lys Asp Ala Ala Glu Val135 140 145 150gtc gcc ttc gcc gag gag ttc ggt ctc ccc atc gcc atc aag gct gcc 1074Val Ala Phe Ala Glu Glu Phe Gly Leu Pro Ile Ala Ile Lys Ala Ala
155 160 165ttc ggt ggc ggc gga cgt ggc atg aag gtc gcc tac gag atg gac gag 1122Phe Gly Gly Gly Gly Arg Gly Met Lys Val Ala Tyr Glu Met Asp Glu
170 175 180gtc gcc gac ctc ttc gaa tcc gcc acc cgt gag gcc acc gcc gcc ttc 1170Val Ala Asp Leu Phe Glu Ser Ala Thr Arg Glu Ala Thr Ala Ala Phe
185 190 195ggt cgt ggt gag tgc ttc gtg gag cgc tac ctg gac aag gcc cgc cac 1218Gly Arg Gly Glu Cys Phe Val Glu Arg Tyr Leu Asp Lys Ala Arg His
200 205 210gtc gag gca cag gtc atc gcc gac aag cac ggc aac gtt gtg gtc gcc 1266Val Glu Ala Gln Val Ile Ala Asp Lys His Gly Asn Val Val Val Ala215 220 225230ggt acc cgt gac tgc tcc ctg cag cgt cgt ttc cag aag ctc gtc gag 1314Gly Thr Arg Asp Cys Ser Leu Gln Arg Arg Phe Gln Lys Leu Val Glu
235 240 245gag gca ccg gca ccg ttc ctc acc gat gag cag cgt gac cgc atc cac 1362Glu Ala Pro Ala Pro Phe Leu Thr Asp Glu Gln Arg Asp Arg Ile His
250 255 260tcc tcc gcc aag gct atc tgc cgc gag gcc ggt tac tac ggt gcc ggc 1410Ser Ser Ala Lys Ala Ile Cys Arg Glu Ala Gly Tyr Tyr Gly Ala Gly
265 270 275acc gtg gag tac ctg gtc ggt tcc gac gga ctg atc tcc ttc ctg gag 1458Thr Val Glu Tyr Leu Val Gly Ser Asp Gly Leu Ile Ser Phe Leu Glu
280 285 290gtc aac acc cgc ctg cag gtg gag cac ccc gtc acc gag gag acc acc 1506Val Asn Thr Arg Leu Gln Val Glu His Pro Val Thr Glu Glu Thr Thr295 300 305 310ggc atc gac ctg gtg cgc gag atg ttc cgc atc gcc gag ggc gcc gag 1554Gly Ile Asp Leu Val Arg Glu Met Phe Arg Ile Ala Glu Gly Ala Glu
315 320 325ctc tcc atc aag gag gac ccg acc cca cgc ggc cac gcc ttc gag ttc 1602Leu Ser Ile Lys Glu Asp Pro Thr Pro Arg Gly His Ala Phe Glu Phe
330 335 340cgc atc aac ggc gag gac gca ggc tcc aac ttc atg ccc gca ccg ggc 1650Arg Ile Asn Gly Glu Asp Ala Gly Ser Asn Phe Met Pro Ala Pro Gly
345 350 355aag atc acc cgc tac cgt gag ccc gcc ggc ccg ggt gtc cgc atg gac 1698Lys Ile Thr Arg Tyr Arg Glu Pro Ala Gly Pro Gly Val Arg Met Asp
360 365 370tcc ggc gtt gtc gag ggt tcc gag atc tcc ggc cag ttc gac tcc atg 1746Ser Gly Val Val Glu Gly Ser Glu Ile Ser Gly Gln Phe Asp Ser Met375 380 385 390ctg gcc aag ctg atc gtc tgg ggc cag acc cgt gag cag gcc ctg gag 1794Leu Ala Lys Leu Ile Val Trp Gly Gln Thr Arg Glu Gln Ala Leu Glu
395 400 405cgt tcc cgt cgt gcg ctc ggc gag tac atc gtc gag ggc atg ccg acc 1842Arg Ser Arg Arg Ala Leu Gly Glu Tyr Ile Val Glu Gly Met Pro Thr
410 415 420gtc atc ccg ttc cac tcc cac atc gtc tcc aac ccg gca ttc gtc ggt 1890Val Ile Pro Phe His Ser His Ile Val Ser Asn Pro Ala Phe Val Gly
425 430 435gac ggc gag ggc ttc gag gtc tac acc aag tgg atc gag gag gtc tgg 1938Asp Gly Glu Gly Phe Glu Val Tyr Thr Lys Trp Ile Glu Glu Val Trp
440 445 450gac aac ccg atc gag ccg ttc gtc gat gca gcc gac ctc gac gac gag 1986Asp Asn Pro Ile Glu Pro Phe Val Asp Ala Ala Asp Leu Asp Asp Glu455 460 465 470gag aag acc ccg tcg cag aag gtc atc gtc gag atc gac ggc cgc cgc 2034Glu Lys Thr Pro Ser Gln Lys Val Ile Val Glu Ile Asp Gly Arg Arg
475 480 485gtc gag gtg gct ctc ccg ggc gac ctc gct ctc ggc ggt ggc gca ggt 2082Val Glu Val Ala Leu Pro Gly Asp Leu Ala Leu Gly Gly Gly Ala Gly
490 495 500gcc gcc aag aag aag ccg aag aag cgt cgc gca ggt ggc gcc aag gcc 2130Ala Ala Lys Lys Lys Pro Lys Lys Arg Arg Ala Gly Gly Ala Lys Ala
505 510 515ggt gtc tcc ggt gac tcc gtc gca gcc ccg atg cag ggc acc gtc atc 2178Gly Val Ser Gly Asp Ser Val Ala Ala Pro Met Gln Gly Thr Val Ile
520 525 530aag gtc aac gtt gag gac ggc gcc gag gtc tcc gag ggt gac acc gtc 2226Lys Val Asn Val Glu Asp Gly Ala Glu Val Ser Glu Gly Asp Thr Val535 540 545 550gtg gtt ctc gag gcc atg aag atg gag aac ccg gtc aag gcc cac aag 2274Val Val Leu Glu Ala Met Lys Met Glu Asn Pro Val Lys Ala His Lys
555 560 565tcc ggt acc gtc tcc ggt ctg acc atc gcc gcg ggt gag ggc gtg acc 2322Ser Gly Thr Val Ser Gly Leu Thr Ile Ala Ala Gly Glu Gly Val Thr
570 575 580aag ggt cag gtt ctc ctg gag atc aag taatcccttc agggaacaga 2369Lys Gly Gln Val Leu Leu Glu Ile Lys
585 590cagccctgtt ct 2381<210>4<211>591<212>PRT<213>Corynebacterium thermoaminogenes<400>4Val Ser Val Glu Thr Arg Lys Ile Thr Lys Val Leu Val Ala Asn Arg 1 5 10 15Gly Glu Ile Ala Ile Arg Val Phe Arg Ala Ala Arg Asp Glu Gly Ile
20 25 30Ala Ser Val Ala Val Tyr Ala Glu Pro Asp Ala Asp Ala Pro Phe Val
35 40 45Glu Tyr Ala Asp Glu Ala Phe Ala Leu Gly Gly Gln Thr Ser Ala Glu
50 55 60Ser Tyr Leu Val Ile Asp Lys Ile Ile Asp Ala Ala Arg Lys Ser Gly 65 70 75 80Ala Asp Ala Val His Pro Gly Tyr Gly Phe Leu Ala Glu Asn Ala Asp
85 90 95Phe Ala Glu Ala Val Ile Asn Glu Gly Leu Ile Trp Ile Gly Pro Ser
100 105 110Pro Glu Ser Ile Arg Ser Leu Gly Asp Lys Val Thr Ala Arg His Ile
115 120 125Ala Asn Asn Ala Asn Ala Pro Met Ala Pro Gly Thr Lys Glu Pro Val
130 135 140Lys Asp Ala Ala Glu Val Val Ala Phe Ala Glu Glu Phe Gly Leu Pro145 150 155 160Ile Ala Ile Lys Ala Ala Phe Gly Gly Gly Gly Arg Gly Met Lys Val
165 170 175Ala Tyr Glu Met Asp Glu Val Ala Asp Leu Phe Glu Ser Ala Thr Arg
180 185 190Glu Ala Thr Ala Ala Phe Gly Arg Gly Glu Cys Phe Val Glu Arg Tyr
195 200 205Leu Asp Lys Ala Arg His Val Glu Ala Gln Val Ile Ala Asp Lys His
210 215 220Gly Asn Val Val Val Ala Gly Thr Arg Asp Cys Ser Leu Gln Arg Arg225 230 235 240Phe Gln Lys Leu Val Glu Glu Ala Pro Ala Pro Phe Leu Thr Asp Glu
245 250 255Gln Arg Asp Arg Ile His Ser Ser Ala Lys Ala Ile Cys Arg Glu Ala
260 265 270Gly Tyr Tyr Gly Ala Gly Thr Val Glu Tyr Leu Val Gly Ser Asp Gly
275 280 285Leu Ile Ser Phe Leu Glu Val Asn Thr Arg Leu Gln Val Glu His Pro
290 295 300Val Thr Glu Glu Thr Thr Gly Ile Asp Leu Val Arg Glu Met Phe Arg305 310 315 320Ile Ala Glu Gly Ala Glu Leu Ser Ile Lys Glu Asp Pro Thr Pro Arg
325 330 335Gly His Ala Phe Glu Phe Arg Ile Asn Gly Glu Asp Ala Gly Ser Asn
340 345 350Phe Met Pro Ala Pro Gly Lys Ile Thr Arg Tyr Arg Glu Pro Ala Gly
355 360 365Pro Gly Val Arg Met Asp Ser Gly Val Val Glu Gly Ser Glu Ile Ser
370 375 380Gly Gln Phe Asp Ser Met Leu Ala Lys Leu Ile Val Trp Gly Gln Thr385 390 395 400Arg Glu Gln Ala Leu Glu Arg Ser Arg Arg Ala Leu Gly Glu Tyr Ile
405 410 415Val Glu Gly Met Pro Thr Val Ile Pro Phe His Ser His Ile Val Ser
420 425 430Asn Pro Ala Phe Val Gly Asp Gly Glu Gly Phe Glu Val Tyr Thr Lys
435 440 445Trp Ile Glu Glu Val Trp Asp Asn Pro Ile Glu Pro Phe Val Asp Ala
450 455 460Ala Asp Leu Asp Asp Glu Glu Lys Thr Pro Ser Gln Lys Val Ile Val465 470 475 480Glu Ile Asp Gly Arg Arg Val Glu Val Ala Leu Pro Gly Asp Leu Ala
485 490 495Leu Gly Gly Gly Ala Gly Ala Ala Lys Lys Lys Pro Lys Lys Arg Arg
500 505 510Ala Gly Gly Ala Lys Ala Gly Val Ser Gly Asp Ser Val Ala Ala Pro
515 520 525Met Gln Gly Thr Val Ile Lys Val Asn Val Glu Asp Gly Ala Glu Val
530 535 540Ser Glu Gly Asp Thr Val Val Val Leu Glu Ala Met Lys Met Glu Asn545 550 555 560Pro Val Lys Ala His Lys Ser Gly Thr Val Ser Gly Leu Thr Ile Ala
565 570 575Ala Gly Glu Gly Val Thr Lys Gly Gln Val Leu Leu Glu Ile Lys
580 585 590<210>5<211>2128<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(339)..(1967)<400>5gctgtcattc cgaccacatt cgccccggga tccgggctcc accacctccc ggacccatgc 60cccatacctg cggaaaccac gggaaacacg ggaaaaaccg atctcattca gaccggcggg 120atccacctgt ggaacagtca gcggcgcggc catggagggc agcgacaggt gacgtccgag 180cacccggttc cccaccgtgg acacggcatt gatccgacac ggtggggata gtttcatgct 240gaaaaactat cgctgtgcag ggaggatccg gaatgtgacc tatttcatgg agaaatgatt 300gtggacgata cccccgggta cggctaccat tccaaaac atg acc att tcc tca cct 356
Met Thr Ile Ser Ser Pro
1 5ttg att gac gtc gct aac ctg cca gac atc aac acc acc gcc ggc aag 404Leu Ile Asp Val Ala Asn Leu Pro Asp Ile Asn Thr Thr Ala Gly Lys
10 15 20atc gcc gac ctg aag gcc cgc cgg gcg gaa gcc cac ttc ccc atg ggt 452Ile Ala Asp Leu Lys Ala Arg Arg Ala Glu Ala His Phe Pro Met Gly
25 30 35gaa aag gcc gta gag aag gtc cac gcg gcc aac cgc ctc acc gcg cgc 500Glu Lys Ala Val Glu Lys Val His Ala Ala Asn Arg Leu Thr Ala Arg
40 45 50gaa cga ctt gac tac ctg ctc gat gaa ggc tcc ttc atc gaa acc gat 548Glu Arg Leu Asp Tyr Leu Leu Asp Glu Gly Ser Phe Ile Glu Thr Asp 55 60 65 70cag ctc gca cgc cac cgc acc acc gcg ttc ggc ctg ggc aac aag cga 596Gln Leu Ala Arg His Arg Thr Thr Ala Phe Gly Leu Gly Asn Lys Arg
75 80 85ccg gcc acc gac ggc atc gtc acc ggc tgg ggc acc atc gac ggc cgc 644Pro Ala Thr Asp Gly Ile Val Thr Gly Trp Gly Thr Ile Asp Gly Arg
90 95 100gag gtc tgc atc ttc tcc cag gac ggc acc gtc ttc ggt ggc gca ctc 692Glu Val Cys Ile Phe Ser Gln Asp Gly Thr Val Phe Gly Gly Ala Leu
105 110 115ggt gag gtc tac ggc gag aag atg atc aag atc atg gag ctg gcc atc 740Gly Glu Val Tyr Gly Glu Lys Met Ile Lys Ile Met Glu Leu Ala Ile
120 125 130gac acc ggc cgc cca ctc atc ggc ctg tac gag ggt gca ggt gcc cgc 788Asp Thr Gly Arg Pro Leu Ile Gly Leu Tyr Glu Gly Ala Gly Ala Arg135 140 145 150atc cag gac ggt gcg gtc tcc ctc gac ttc atc tcc cag acc ttc tat 836Ile Gln Asp Gly Ala Val Ser Leu Asp Phe Ile Ser Gln Thr Phe Tyr
155 160 165cag aac atc cag gcc tcc ggc gtg atc ccg cag atc tcc gtg atc atg 884Gln Asn Ile Gln Ala Ser Gly Val Ile Pro Gln Ile Ser Val Ile Met
170 175 180ggt gcc tgc gcc ggt ggc aac gcc tac ggc ccg gcc ctg acc gac ttc 932Gly Ala Cys Ala Gly Gly Asn Ala Tyr Gly Pro Ala Leu Thr Asp Phe
185 190 195gtg gtc atg gtg gac aag acc tcg aag atg ttc gtc acc ggc ccc gat 980Val Val Met Val Asp Lys Thr Ser Lys Met Phe Val Thr Gly Pro Asp
200 205 210gtg atc aag acc gtc acc ggc gag gag atc acc cag gag gag ctc ggc 1028Val Ile Lys Thr Val Thr Gly Glu Glu Ile Thr Gln Glu Glu Leu Gly215 220 225 230gga gca acc acc cac atg gtc acc gcc ggc aac tcc cac tac acc gtc 1076Gly Ala Thr Thr His Met Val Thr Ala Gly Asn Ser His Tyr Thr Val
235 240 245gcc acc gat gag gag gcc ctc gac tgg gtc cag gac ctc atc tcc ttc 1124Ala Thr Asp Glu Glu Ala Leu Asp Trp Val Gln Asp Leu Ile Ser Phe
250 255 260ctg ccc tcc aac aat cgc tcc tac gcc ccg gtg gag gag ttc gac gag 1172Leu Pro Ser Asn Asn Arg Ser Tyr Ala Pro Val Glu Glu Phe Asp Glu
265 270 275gag gac ggt ggc atc gcc gag aac atc acc gcc gat gac ctg aag ctg 1220Glu Asp Gly Gly Ile Ala Glu Asn Ile Thr Ala Asp Asp Leu Lys Leu
280 285 290gat gag atc atc ccg gat tcc gcc acc gtg ccc tat gat gtc cgc gac 1268Asp Glu Ile Ile Pro Asp Ser Ala Thr Val Pro Tyr Asp Val Arg Asp295 300 305 310gtc atc cag tgc ctg acc gac gac ggt gag tac ctg gag atc cag gcc 1316Val Ile Gln Cys Leu Thr Asp Asp Gly Glu Tyr Leu Glu Ile Gln Ala
315 320 325gac cga gcc gag aat gtc gtc atc gcc ttc ggc cgc atc gag ggc cag 1364Asp Arg Ala Glu Asn Val Val Ile Ala Phe Gly Arg Ile Glu Gly Gln
330 335 340tcc gtc ggt ttc gtc gcc aac cag ccg acc cag ttc gcc ggc tgc ctg 1412Ser Val Gly Phe Val Ala Asn Gln Pro Thr Gln Phe Ala Gly Cys Leu
345 350 355gac atc gac tcc tcc gag aag gca gcc cgc ttc gtc cgc acc tgc gat 1460Asp Ile Asp Ser Ser Glu Lys Ala Ala Arg Phe Val Arg Thr Cys Asp
360 365 370gcc ttc aac atc ccg atc gtc atg ctt gtc gac gtc ccc ggc ttc ctc 1508Ala Phe Asn Ile Pro Ile Val Met Leu Val Asp Val Pro Gly Phe Leu375 380 385 390ccc ggt gcc ggc cag gag tac ggc ggc atc ctg cgt cgt ggc gcc aaa 1556Pro Gly Ala Gly Gln Glu Tyr Gly Gly Ile Leu Arg Arg Gly Ala Lys
395 400 405ctg ctc tac gcc tac ggt gag gcc acc gtc ccg aag atc acc gtg acc 1604Leu Leu Tyr Ala Tyr Gly Glu Ala Thr Val Pro Lys Ile Thr Val Thr
410 415 420atg cgc aag gcc tac ggc ggt gcg tac tgt gtc atg gga tcc aag ggt 1652Met Arg Lys Ala Tyr Gly Gly Ala Tyr Cys Val Met Gly Ser Lys Gly
425 430 435ctg ggc gca gac atc aac ctg gcc tgg ccg acc gcg cag atc gcc gtc 1700Leu Gly Ala Asp Ile Asn Leu Ala Trp Pro Thr Ala Gln Ile Ala Val
440 445 450atg ggt gcc gcc ggc gcg gtc cag ttc atc tac cgc aag gag ctc atg 1748Met Gly Ala Ala Gly Ala Val Gln Phe Ile Tyr Arg Lys Glu Leu Met455 460 465 470gcc gct gat gcc aag ggc ctg gac acc gtc gcc ctg gcc cag tcc ttc 1796Ala Ala Asp Ala Lys Gly Leu Asp Thr Val Ala Leu Ala Gln Ser Phe
475 480 485gag cgt gag tac gag gac cac atg ctc aac ccg tac ctg gcg gcc gag 1844Glu Arg Glu Tyr Glu Asp His Met Leu Asn Pro Tyr Leu Ala Ala Glu
490 495 500cgt ggc ctc atc gac gcg gtg atc ctg ccg tcc gag acc cgt ggc cag 1892Arg Gly Leu Ile Asp Ala Val Ile Leu Pro Ser Glu Thr Arg Gly Gln
505 510 515atc gca cgc aac ctg cgt ctg ctc aag cac aag aat gtc tcc cgc cct 1940Ile Ala Arg Asn Leu Arg Leu Leu Lys His Lys Asn Val Ser Arg Pro
520 525 530gcc cgc aag cac ggc aac atg cca ctg taagcacccg ggaccacccc 1987Ala Arg Lys His Gly Asn Met Pro Leu535 540ctacgcccgc acccacggcc ctttgctggc aggtgcgggc gctgtgcgtt ttccgcgcct 2047gccgacgccc ggccccctgc cctgtgatgc gatctgcgga tgtgatctgc gcccgcgcca 2107actcccctgg ttgaaccctg c 2128<210>6<211>543<212>PRT<213>Corynebacterium thermoaminogenes<400>6Met Thr Ile Ser Ser Pro Leu Ile Asp Val Ala Asn Leu Pro Asp Ile 1 5 10 15Asn Thr Thr Ala Gly Lys Ile Ala Asp Leu Lys Ala Arg Arg Ala Glu
20 25 30Ala His Phe Pro Met Gly Glu Lys Ala Val Glu Lys Val His Ala Ala
35 40 45Asn Arg Leu Thr Ala Arg Glu Arg Leu Asp Tyr Leu Leu Asp Glu Gly
50 55 60Ser Phe Ile Glu Thr Asp Gln Leu Ala Arg His Arg Thr Thr Ala Phe 65 70 75 80Gly Leu Gly Asn Lys Arg Pro Ala Thr Asp Gly Ile Val Thr Gly Trp
85 90 95Gly Thr Ile Asp Gly Arg Glu Val Cys Ile Phe Ser Gln Asp Gly Thr
100 105 110Val Phe Gly Gly Ala Leu Gly Glu Val Tyr Gly Glu Lys Met Ile Lys
115 120 125Ile Met Glu Leu Ala Ile Asp Thr Gly Arg Pro Leu Ile Gly Leu Tyr
130 135 140Glu Gly Ala Gly Ala Arg Ile Gln Asp Gly Ala Val Ser Leu Asp Phe145 150 155 160Ile Ser Gln Thr Phe Tyr Gln Asn Ile Gln Ala Ser Gly Val Ile Pro
165 170 175Gln Ile Ser Val Ile Met Gly Ala Cys Ala Gly Gly Asn Ala Tyr Gly
180 185 190Pro Ala Leu Thr Asp Phe Val Val Met Val Asp Lys Thr Ser Lys Met
195 200 205Phe Val Thr Gly Pro Asp Val Ile Lys Thr Val Thr Gly Glu Glu Ile
210 215 220Thr Gln Glu Glu Leu Gly Gly Ala Thr Thr His Met Val Thr Ala Gly225 230 235 240Asn Ser His Tyr Thr Val Ala Thr Asp Glu Glu Ala Leu Asp Trp Val
245 250 255Gln Asp Leu Ile Ser Phe Leu Pro Ser Asn Asn Arg Ser Tyr Ala Pro
260 265 270Val Glu Glu Phe Asp Glu Glu Asp Gly Gly Ile Ala Glu Asn Ile Thr
275 280 285Ala Asp Asp Leu Lys Leu Asp Glu Ile Ile Pro Asp Ser Ala Thr Val
290 295 300Pro Tyr Asp Val Arg Asp Val Ile Gln Cys Leu Thr Asp Asp Gly Glu305 310 315 320Tyr Leu Glu Ile Gln Ala Asp Arg Ala Glu Asn Val Val Ile Ala Phe
325 330 335Gly Arg Ile Glu Gly Gln Ser Val Gly Phe Val Ala Asn Gln Pro Thr
340 345 350Gln Phe Ala Gly Cys Leu Asp Ile Asp Ser Ser Glu Lys Ala Ala Arg
355 360 365Phe Val Arg Thr Cys Asp Ala Phe Asn Ile Pro Ile Val Met Leu Val
370 375 380Asp Val Pro Gly Phe Leu Pro Gly Ala Gly Gln Glu Tyr Gly Gly Ile385 390 395 400Leu Arg Arg Gly Ala Lys Leu Leu Tyr Ala Tyr Gly Glu Ala Thr Val
405 410 415Pro Lys Ile Thr Val Thr Met Arg Lys Ala Tyr Gly Gly Ala Tyr Cys
420 425 430Val Met Gly Ser Lys Gly Leu Gly Ala Asp Ile Asn Leu Ala Trp Pro
435 440 445Thr Ala Gln Ile Ala Val Met Gly Ala Ala Gly Ala Val Gln Phe Ile
450 455 460Tyr Arg Lys Glu Leu Met Ala Ala Asp Ala Lys Gly Leu Asp Thr Val465 470 475 480Ala Leu Ala Gln Ser Phe Glu Arg Glu Tyr Glu Asp His Met Leu Asn
485 490 495Pro Tyr Leu Ala Ala Glu Arg Gly Leu Ile Asp Ala Val Ile Leu Pro
500 505 510Ser Glu Thr Arg Gly Gln Ile Ala Arg Asn Leu Arg Leu Leu Lys His
515520 525Lys Asn Val Ser Arg Pro Ala Arg Lys His Gly Asn Met Pro Leu
530 535 540<210>7<211>2076<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(412)..(2022)<400>7acgcccggcc ccctgccctg tgatgcgatc tgcggatgtg atctgcgccc gcgccaactc 60ccctggttga accctgccac ataccctgag tcgcacctgg gtggggtcac tttccacctc 120acggggggga ggaggtcaca taggccatac gctgcacttt tgatgaagtg tgggcagatc 180gaccgggcaa atctgggaaa taaggggcct ggtgaactag cattcccctt agcgaagggt 240gagcatcgcg gaccccgcga tgtcccaacc ggtcgtaaat tcatgtgccg ccacagtccc 300ctcaccaggg gatcggaacc agcccagcct gattccggcg tgacggacct caccgtgaac 360aagtccccgc attactcaca gaactcacac caggatttag actaagaaac c atg act 417
Met Thr
1gca gca acg aca gca cct gat ctg acc acc acc gcc ggc aaa ctc gcg 465Ala Ala Thr Thr Ala Pro Asp Leu Thr Thr Thr Ala Gly Lys Leu Ala
5 10 15gat ctc cgc gcc cgc ctt tcc gag acc cag gcc ccc atg ggt cag gcc 513Asp Leu Arg Ala Arg Leu Ser Glu Thr Gln Ala Pro Met Gly Gln Ala
20 25 30tcc gtg gag aag gtg cac gag gca ggg aag aag acc gca cgc gag cgc 561Ser Val Glu Lys Val His Glu Ala Gly Lys Lys Thr Ala Arg Glu Arg 35 40 45 50atc gag tac ctg ctc gat gag ggc tcc ttc gtt gag gtc gat gcc ctc 609Ile Glu Tyr Leu Leu Asp Glu Gly Ser Phe Val Glu Val Asp Ala Leu
55 60 65gcc cgc cac cgt tcc aag aac ttc ggc ctg gac tcc aag cgc ccg gtc 657Ala Arg His Arg Ser Lys Asn Phe Gly Leu Asp Ser Lys Arg Pro Val
70 75 80acc gac ggt gtg gtc acc ggt tac ggc acc atc gac gga cgc aag gtc 705Thr Asp Gly Val Val Thr Gly Tyr Gly Thr Ile Asp Gly Arg Lys Val
85 90 95tgc gtc ttc tcc cag gac ggc gct atc ttc ggc ggt gcc ctc ggt gag 753Cys Val Phe Ser Gln Asp Gly Ala Ile Phe Gly Gly Ala Leu Gly Glu
100 105 110gtc tac ggc gag aag atc gtc aag atc atg gac ctg gcc atc aag acc 801Val Tyr Gly Glu Lys Ile Val Lys Ile Met Asp Leu Ala Ile Lys Thr115 120 125 130ggt gtc ccc ctc atc ggc atc aac gag ggc gcc ggc gcc cgc atc cag 849Gly Val Pro Leu Ile Gly Ile Asn Glu Gly Ala Gly Ala Arg Ile Gln
135 140 145gaa ggc gtt gtc tcc ctg ggc ctg tac tcc cag atc ttc tac cgc aac 897Glu Gly Val Val Ser Leu Gly Leu Tyr Ser Gln Ile Phe Tyr Arg Asn
150 155 160acc cag gca tcc ggt gtc atc cca cag atc tcc ctc atc atg ggt gcc 945Thr Gln Ala Ser Gly Val Ile Pro Gln Ile Ser Leu Ile Met Gly Ala
165 170 175tgc gcc ggt ggc cat gtg tac tcc ccc gcc ctg acc gac ttc atc atc 993Cys Ala Gly Gly His Val Tyr Ser Pro Ala Leu Thr Asp Phe Ile Ile
180 185 190atg gtg gac aag acc tcc aag atg ttc atc acc ggc ccc gac gtg atc 1041Met Val Asp Lys Thr Ser Lys Met Phe Ile Thr Gly Pro Asp Val Ile195 200 205 210aag acc gtc acc ggc gag gag gtc acc cag gag gaa ctg ggt ggt gcc 1089Lys Thr Val Thr Gly Glu Glu Val Thr Gln Glu Glu Leu Gly Gly Ala
215 220 225tac acc cac atg gcc cag tcc ggc acc tcg cac tac acc gca gcc gat 1137Tyr Thr His Met Ala Gln Ser Gly Thr Ser His Tyr Thr Ala Ala Asp
230 235 240gac tcc gat gcc ctc gac tgg gtc cgt gag ctg gtc agc tac ctg ccg 1185Asp Ser Asp Ala Leu Asp Trp Val Arg Glu Leu Val Ser Tyr Leu Pro
245 250 255tcc aac aac cgt gcg gag acc cca cgc cag gac gcc gac atc atg gtg 1233Ser Asn Asn Arg Ala Glu Thr Pro Arg Gln Asp Ala Asp Ile Met Val
260 265 270ggc tcc atc aag gag aac atc acc gag acc gac ctc gaa ctc gac acc 1281Gly Ser Ile Lys Glu Asn Ile Thr Glu Thr Asp Leu Glu Leu Asp Thr275 280 285 290ctg atc ccg gat tcc ccg aac cag ccg tac gac atg aag gac gtc atc 1329Leu Ile Pro Asp Ser Pro Asn Gln Pro Tyr Asp Met Lys Asp Val Ile
295 300 305acc cgc atc gtc gat gat gcc gag ttc ttc gag atc cag gag ggt tac 1377Thr Arg Ile Val Asp Asp Ala Glu Phe Phe Glu Ile Gln Glu Gly Tyr
310 315 320gcc gag aac atc atc tgc ggt ttc gcc cgc gtc gag ggt cgt gcc gtg 1425Ala Glu Asn Ile Ile Cys Gly Phe Ala Arg Val Glu Gly Arg Ala Val
325 330 335ggt atc gtg gcc aac cag ccg atg cag ttc gcc ggc tgc ctg gac atc 1473Gly Ile Val Ala Asn Gln Pro Met Gln Phe Ala Gly Cys Leu Asp Ile
340 345 350aag gca tcc gag aag gcc gcc cgc ttc atc cgc acc tgt gac gcc ttc 1521Lys Ala Ser Glu Lys Ala Ala Arg Phe Ile Arg Thr Cys Asp Ala Phe355 360 365 370aac atc ccg atc atc gag ctt gtc gac gtc cca ggc ttc ctc ccg ggc 1569Asn Ile Pro Ile Ile Glu Leu Val Asp Val Pro Gly Phe Leu Pro Gly
375 380 385acc aac cag gag ttc gac ggc atc atc cgt cgc ggc gcg aag ctg ctc 1617Thr Asn Gln Glu Phe Asp Gly Ile Ile Arg Arg Gly Ala Lys Leu Leu
390 395 400tac gcc tac gcc gag gcc acc gtc ggc aag atc acc gtg atc acc cgc 1665Tyr Ala Tyr Ala Glu Ala Thr Val Gly Lys Ile Thr Val Ile Thr Arg
405 410 415aag tcc tac ggc ggt gcc tac tgc gtg atg ggc tcc aag gac atg ggt 1713Lys Ser Tyr Gly Gly Ala Tyr Cys Val Met Gly Ser Lys Asp Met Gly
420 425 430gcg gac ctc gtc ttc gca tgg ccc acc gcg cag atc gcc gtc atg ggt 1761Ala Asp Leu Val Phe Ala Trp Pro Thr Ala Gln Ile Ala Val Met Gly435 440 445 450gcc tcc ggt gcc gtc ggc ttc atc tac cgc aag gag ctc aag cag gct 1809Ala Ser Gly Ala Val Gly Phe Ile Tyr Arg Lys Glu Leu Lys Gln Ala
455 460 465gca gcg gcc ggc gag gat gtc acc gcg ctg atg aag aag tac gag cag 1857Ala Ala Ala Gly Glu Asp Val Thr Ala Leu Met Lys Lys Tyr Glu Gln
470 475 480gag tac gag gag acc ctg gtc aac ccg tac atg gct gca gag cgt ggc 1905Glu Tyr Glu Glu Thr Leu Val Asn Pro Tyr Met Ala Ala Glu Arg Gly
485 490 495tac gtc gac gcc gtc atc cca cca tcc gag acc cgt ggt cag atc atc 1953Tyr Val Asp Ala Val Ile Pro Pro Ser Glu Thr Arg Gly Gln Ile Ile
500 505 510gag ggt ctg cgt ctg ctc gac cgc aag gtg gtc aac gtc ccg gcc aag 2001Glu Gly Leu Arg Leu Leu Asp Arg Lys Val Val Asn Val Pro Ala Lys515 520 525 530aag cac ggt aac atc ccg ctg taaaccgtct tcccctccgg caccacgccg 2052Lys His Gly Asn Ile Pro Leu
535gagaaggctt tgtccgcagc tgtc2076<210>8<211>537<212>PRT<213>Corynebacterium thermoaminogenes<400>8Met Thr Ala Ala Thr Thr Ala Pro Asp Leu Thr Thr Thr Ala Gly Lys 1 5 10 15Leu Ala Asp Leu Arg Ala Arg Leu Ser Glu Thr Gln Ala Pro Met Gly
20 25 30Gln Ala Ser Val Glu Lys Val His Glu Ala Gly Lys Lys Thr Ala Arg
35 40 45Glu Arg Ile Glu Tyr Leu Leu Asp Glu Gly Ser Phe Val Glu Val Asp
50 55 60Ala Leu Ala Arg His Arg Ser Lys Asn Phe Gly Leu Asp Ser Lys Arg 65 70 75 80Pro Val Thr Asp Gly Val Val Thr Gly Tyr Gly Thr Ile Asp Gly Arg
85 90 95Lys Val Cys Val Phe Ser Gln Asp Gly Ala Ile Phe Gly Gly Ala Leu
100 105 110Gly Glu Val Tyr Gly Glu Lys Ile Val Lys Ile Met Asp Leu Ala Ile
115 120 125Lys Thr Gly Val Pro Leu Ile Gly Ile Asn Glu Gly Ala Gly Ala Arg
130 135 140Ile Gln Glu Gly Val Val Ser Leu Gly Leu Tyr Ser Gln Ile Phe Tyr145 150 155 160Arg Asn Thr Gln Ala Ser Gly Val Ile Pro Gln Ile Ser Leu Ile Met
165 170 175Gly Ala Cys Ala Gly Gly His Val Tyr Ser Pro Ala Leu Thr Asp Phe
180 185 190Ile Ile Met Val Asp Lys Thr Ser Lys Met Phe Ile Thr Gly Pro Asp
195 200 205Val Ile Lys Thr Val Thr Gly Glu Glu Val Thr Gln Glu Glu Leu Gly
210 215 220Gly Ala Tyr Thr His Met Ala Gln Ser Gly Thr Ser His Tyr Thr Ala225230 235 240Ala Asp Asp Ser Asp Ala Leu Asp Trp Val Arg Glu Leu Val Ser Tyr
245 250 255Leu Pro Ser Asn Asn Arg Ala Glu Thr Pro Arg Gln Asp Ala Asp Ile
260 265 270Met Val Gly Ser Ile Lys Glu Asn Ile Thr Glu Thr Asp Leu Glu Leu
275 280 285Asp Thr Leu Ile Pro Asp Ser Pro Asn Gln Pro Tyr Asp Met Lys Asp
290 295 300Val Ile Thr Arg Ile Val Asp Asp Ala Glu Phe Phe Glu Ile Gln Glu305 310 315 320Gly Tyr Ala Glu Asn Ile Ile Cys Gly Phe Ala Arg Val Glu Gly Arg
325 330 335Ala Val Gly Ile Val Ala Asn Gln Pro Met Gln Phe Ala Gly Cys Leu
340 345 350Asp Ile Lys Ala Ser Glu Lys Ala Ala Arg Phe Ile Arg Thr Cys Asp
355 360 365Ala Phe Asn Ile Pro Ile Ile Glu Leu Val Asp Val Pro Gly Phe Leu
370 375 380Pro Gly Thr Asn Gln Glu Phe Asp Gly Ile Ile Arg Arg Gly Ala Lys385 390 395 400Leu Leu Tyr Ala Tyr Ala Glu Ala Thr Val Gly Lys Ile Thr Val Ile
405 410 415Thr Arg Lys Ser Tyr Gly Gly Ala Tyr Cys Val Met Gly Ser Lys Asp
420 425 430Met Gly Ala Asp Leu Val Phe Ala Trp Pro Thr Ala Gln Ile Ala Val
435 440 445Met Gly Ala Ser Gly Ala Val Gly Phe Ile Tyr Arg Lys Glu Leu Lys
450 455 460Gln Ala Ala Ala Ala Gly Glu Asp Val Thr Ala Leu Met Lys Lys Tyr465 470 475 480Glu Gln Glu Tyr Glu Glu Thr Leu Val Asn Pro Tyr Met Ala Ala Glu
485 490 495Arg Gly Tyr Val Asp Ala Val Ile Pro Pro Ser Glu Thr Arg Gly Gln
500 505 510Ile Ile Glu Gly Leu Arg Leu Leu Asp Arg Lys Val Val Asn Val Pro
515 520 525Ala Lys Lys His Gly Asn Ile Pro Leu
530 535<210>9<211>1643<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(326)..(1363)<400>9agcgcgccgg cagccaccag tgggatcgtg cccagcggac ggatgccgga ttcacggcgg 60tcagccaccc gccgatgaga cctgcagcga caacggtggc ggtgctgacc tggtcagcgt 120ctttgagttt catatccatg tcagacagtc taaccactct ctccgacgcg tccgaacatg 180ctggggtggc ggacaccatg tccgttcggg cgttgccccg acgggggaaa atcgcaggca 240gatgtgtccg atgtgggata aacccaccgg ttcgggcgtg tcttcgggat caatggcaca 300gcattaaccg tgtggggggt ttaat atg gga gcc atg cga att gcc act ctc 352
Met Gly Ala Met Arg Ile Ala Thr Leu
1 5acg tca ggc ggc gac tgc ccc gga ctc aat gct gtc atc agg gga atc 400Thr Ser Gly Gly Asp Cys Pro Gly Leu Asn Ala Val Ile Arg Gly Ile 10 15 20 25gtc cgt acc gca agt aat gaa ttc ggt tcc acc gtc gtg ggt tat cag 448Val Arg Thr Ala Ser Asn Glu Phe Gly Ser Thr Val Val Gly Tyr Gln
30 35 40gac ggc tgg gag ggc ctg ctg gcg gac cga cgt gtt cag ctc tat gac 496Asp Gly Trp Glu Gly Leu Leu Ala Asp Arg Arg Val Gln Leu Tyr Asp
45 50 55gat gag gac atc gac cgc atc ctg ctc cgc ggt gga aca atc ctg ggc 544Asp Glu Asp Ile Asp Arg Ile Leu Leu Arg Gly Gly Thr Ile Leu Gly
60 65 70acc ggt cgt ctc cac ccc gac aag ttc aga gcc gga atc gac cag gtc 592Thr Gly Arg Leu His Pro Asp Lys Phe Arg Ala Gly Ile Asp Gln Val
75 80 85aag gcg aat ctc gcc gat gcg gga att gac gca ctc atc ccg atc ggt 640Lys Ala Asn Leu Ala Asp Ala Gly Ile Asp Ala Leu Ile Pro Ile Gly90 95 100 105ggc gag ggc acc ctc aag gga gcg aag tgg ctc gcc gac aac ggc atc 688Gly Glu Gly Thr Leu Lys Gly Ala Lys Trp Leu Ala Asp Asn Gly Ile
110 115 120ccc gtg gtc ggt gtc ccg aaa acc atc gac aat gat gtc aac ggc acg 736Pro Val Val Gly Val Pro Lys Thr Ile Asp Asn Asp Val Asn Gly Thr
125 130 135gat ttc acc ttc ggt ttc gat tcc gcg gtc tct gtg gcc acc gac gcc 784Asp Phe Thr Phe Gly Phe Asp Ser Ala Val Ser Val Ala Thr Asp Ala
140 145 150atc gac cgg ctg cac acc acg gcg gaa tcc cac aac cgt gtg atg atc 832Ile Asp Arg Leu His Thr Thr Ala Glu Ser His Asn Arg Val Met Ile
155 160 165gtc gag gtc atg ggc cgc cac gtc ggt tgg atc gca ctg cat gcc ggc 880Val Glu Val Met Gly Arg His Val Gly Trp Ile Ala Leu His Ala Gly170 175 180 185atg gcc ggt gga gcc cac tac acc gtc atc ccc gag gtg ccc ttc gac 928Met Ala Gly Gly Ala His Tyr Thr Val Ile Pro Glu Val Pro Phe Asp
190 195 200atc tcg gag atc tgc aag cgt atg gaa cgt cgc ttc cag atg ggg gag 976Ile Ser Glu Ile Cys Lys Arg Met Glu Arg Arg Phe Gln Met Gly Glu
205 210 215aag tac ggc atc atc gtc gtc gcg gag ggt gcc ctg ccc aag gag gga 1024Lys Tyr Gly Ile Ile Val Val Ala Glu Gly Ala Leu Pro Lys Glu Gly
220 225 230acc atg gag ctg cgt gag ggg gag gtg gat cag ttc ggt cac aag acc 1072Thr Met Glu Leu Arg Glu Gly Glu Val Asp Gln Phe Gly His Lys Thr
235 240 245ttc acc ggc atc ggc cag cag atc gcc gac gag gtg cac agg cgt ctg 1120Phe Thr Gly Ile Gly Gln Gln Ile Ala Asp Glu Val His Arg Arg Leu250 255 260 265ggt cat gat gtc cgc acc acg gtc ctg ggc cat atc cag cgt ggt ggc 1168Gly His Asp Val Arg Thr Thr Val Leu Gly His Ile Gln Arg Gly Gly
270 275 280acc ccc acc gcc ttc gac cgt gtc ctg gcc acc cgg tac ggt gtc cgc 1216Thr Pro Thr Ala Phe Asp Arg Val Leu Ala Thr Arg Tyr Gly Val Arg
285 290 295gcc gcg cgt gcc tgc cac gag ggt cag ttc aac acc gtg gtc gcg ctc 1264Ala Ala Arg Ala Cys His Glu Gly Gln Phe Asn Thr Val Val Ala Leu
300 305 310aag ggg gag cgc atc cgg atg atc tcc ttc gat gag gcc gtg ggc acc 1312Lys Gly Glu Arg Ile Arg Met Ile Ser Phe Asp Glu Ala Val Gly Thr
315 320 325ctg aag aag gtg ccg atg gaa cgc tgg gtg acc gcc cag gct atg ttc 1360Leu Lys Lys Val Pro Met Glu Arg Trp Val Thr Ala Gln Ala Met Phe330 335 340 345ggt tagtcaggcc gcattcccgg ttccgcgccc gcggggccgg gttttttcat1413Glygccccggaac acatcggtat gaaatcgtga tatgcattac ttgacgggga agtgggggat 1473ccgtcacctc gcgttgtcca actacagccc gcagcgcctg cgggaattct tcgagcaatc 1533cgccgattcc ccggcccgtc ccgtcgccgt ccaaccgcag tacaatctgc tggcccgccg 1593ggattatgag accggtatcc gcccggtcgt ggacgagttc ggtcccgcgg1643<210>10<211>346<212>PRT<213>Corynebacterium thermoaminogenes<400>10Met Gly Ala Met Arg Ile Ala Thr Leu Thr Ser Gly Gly Asp Cys Pro 1 5 10 15Gly Leu Asn Ala Val Ile Arg Gly Ile Val Arg Thr Ala Ser Asn Glu
20 25 30Phe Gly Ser Thr Val Val Gly Tyr Gln Asp Gly Trp Glu Gly Leu Leu
35 40 45Ala Asp Arg Arg Val Gln Leu Tyr Asp Asp Glu Asp Ile Asp Arg Ile
50 55 60Leu Leu Arg Gly Gly Thr Ile Leu Gly Thr Gly Arg Leu His Pro Asp 65 70 75 80Lys Phe Arg Ala Gly Ile Asp Gln Val Lys Ala Asn Leu Ala Asp Ala
85 90 95Gly Ile Asp Ala Leu Ile Pro Ile Gly Gly Glu Gly Thr Leu Lys Gly
100 105 110Ala Lys Trp Leu Ala Asp Asn Gly Ile Pro Val Val Gly Val Pro Lys
115 120 125Thr Ile Asp Asn Asp Val Asn Gly Thr Asp Phe Thr Phe Gly Phe Asp
130 135 140Ser Ala Val Ser Val Ala Thr Asp Ala Ile Asp Arg Leu His Thr Thr145 150 155 160Ala Glu Ser His Asn Arg Val Met Ile Val Glu Val Met Gly Arg His
165 170 175Val Gly Trp Ile Ala Leu His Ala Gly Met Ala Gly Gly Ala His Tyr
180 185 190Thr Val Ile Pro Glu Val Pro Phe Asp Ile Ser Glu Ile Cys Lys Arg
195 200 205Met Glu Arg Arg Phe Gln Met Gly Glu Lys Tyr Gly Ile Ile Val Val
210 215 220Ala Glu Gly Ala Leu Pro Lys Glu Gly Thr Met Glu Leu Arg Glu Gly225 230 235 240Glu Val Asp Gln Phe Gly His Lys Thr Phe Thr Gly Ile Gly Gln Gln
245 250 255Ile Ala Asp Glu Val His Arg Arg Leu Gly His Asp Val Arg Thr Thr
260 265 270Val Leu Gly His Ile Gln Arg Gly Gly Thr Pro Thr Ala Phe Asp Arg
275 280 285Val Leu Ala Thr Arg Tyr Gly Val Arg Ala Ala Arg Ala Cys His Glu
290 295 300Gly Gln Phe Asn Thr Val Val Ala Leu Lys Gly Glu Arg Ile Arg Met305 310 315 320Ile Ser Phe Asp Glu Ala Val Gly Thr Leu Lys Lys Val Pro Met Glu
325 330 335Arg Trp Val Thr Ala Gln Ala Met Phe Gly
340 345<210>11<211>498<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(1)..(498)<400>11tac tac cag cac gat cca ggt ttc ccc ttc gca cca aag cgc acc ggt 48Tyr Tyr Gln His Asp Pro Gly Phe Pro Phe Ala Pro Lys Arg Thr Gly 1 5 10 15tgg gct cac acc acc acg ccg ttg acc gga ccg cag cga ttg cag tgg 96Trp Ala His Thr Thr Thr Pro Leu Thr Gly Pro Gln Arg Leu Gln Trp
20 25 30acg cac ctg ccc gat gct ctt tac ccg gat gta tcc tat gac ctg gat 144Thr His Leu Pro Asp Ala Leu Tyr Pro Asp Val Ser Tyr Asp Leu Asp
35 40 45gga tgc tat tcc ggc gga gcc gta ttt tct gac ggc acg ctt aaa ctt 192Gly Cys Tyr Ser Gly Gly Ala Val Phe Ser Asp Gly Thr Leu Lys Leu
50 55 60ttc tac acc ggc aac cga aaa att gac ggc aag cgc cgc gcc acc caa 240Phe Tyr Thr Gly Asn Arg Lys Ile Asp Gly Lys Arg Arg Ala Thr Gln 65 70 75 80aac ctc gtc gaa gtc gag gac cca act ggg ctg atg ggc ggc att cat 288Asn Leu Val Glu Val Glu Asp Pro Thr Gly Leu Met Gly Gly Ile His
85 90 95cgc cgc tcg cct aaa aat ccg ctt atc gac gga ccc gcc agc ggt ttt 336Arg Arg Ser Pro Lys Asn Pro Leu Ile Asp Gly Pro Ala Ser Gly Phe
100 105 110acg ccc cac tac cgc gat ccc atg atc agc cct gat ggg gat ggt tgg 384Thr Pro His Tyr Arg Asp Pro Met Ile Ser Pro Asp Gly Asp Gly Trp
115 120 125aag atg gtt ctt ggg gct cag cgc gaa aac ctc acc ggt gca gcg gtt 432Lys Met Val Leu Gly Ala Gln Arg Glu Asn Leu Thr Gly Ala Ala Val
130 135 140cta tac cgc tcg gca gat ctt gaa aac tgg gaa ttc tcc ggt gaa atc 480Leu Tyr Arg Ser Ala Asp Leu Glu Asn Trp Glu Phe Ser Gly Glu Ile145 150 155 160acc ttt gac ctc agc gac 498Thr Phe Asp Leu Ser Asp
165<210>12<211>166<212>PRT<213>Corynebacterium thermoaminogenes<400>12Tyr Tyr Gln His Asp Pro Gly Phe Pro Phe Ala Pro Lys Arg Thr Gly 1 5 10 15Trp Ala His Thr Thr Thr Pro Leu Thr Gly Pro Gln Arg Leu Gln Trp
20 25 30Thr His Leu Pro Asp Ala Leu Tyr Pro Asp Val Ser Tyr Asp Leu Asp
35 40 45Gly Cys Tyr Ser Gly Gly Ala Val Phe Ser Asp Gly Thr Leu Lys Leu
50 55 60Phe Tyr Thr Gly Asn Arg Lys Ile Asp Gly Lys Arg Arg Ala Thr Gln 65 70 75 80Asn Leu Val Glu Val Glu Asp Pro Thr Gly Leu Met Gly Gly Ile His
85 90 95Arg Arg Ser Pro Lys Asn Pro Leu Ile Asp Gly Pro Ala Ser Gly Phe
100 105 110Thr Pro His Tyr Arg Asp Pro Met Ile Ser Pro Asp Gly Asp Gly Trp
115 120 125Lys Met Val Leu Gly Ala Gln Arg Glu Asn Leu Thr Gly Ala Ala Val
130 135 140Leu Tyr Arg Ser Ala Asp Leu Glu Asn Trp Glu Phe Ser Gly Glu Ile145 150 155 160Thr Phe Asp Leu Ser Asp
165<210>13<211>479<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(1)..(477)<400>13tac tac cag cac gat cca ggt ttc ccc ttc gca cca aag cgc acc ggc 48Tyr Tyr Gln His Asp Pro Gly Phe Pro Phe Ala Pro Lys Arg Thr Gly 1 5 10 15tgg gct cac acc acc acg ccg ttg acc gga ccg cag cga ttg cag tgg 96Trp Ala His Thr Thr Thr Pro Leu Thr Gly Pro Gln Arg Leu Gln Trp
20 25 30acg cac ctg ccc gac gct ctt tac ccg gat gca tcc tat gac ctg gat 144Thr His Leu Pro Asp Ala Leu Tyr Pro Asp Ala Ser Tyr Asp Leu Asp
35 40 45gga tgc tat tcc ggt gga gcc gta ttt act gac ggc aca ctt aaa ctt 192Gly Cys Tyr Ser Gly Gly Ala Val Phe Thr Asp Gly Thr Leu Lys Leu
50 55 60ttc tac acc ggc aac cta aaa att gac ggc aag cgc cgc gcc acc caa 240Phe Tyr Thr Gly Asn Leu Lys Ile Asp Gly Lys Arg Arg Ala Thr Gln 65 70 75 80aac ctc gtc gaa gtc gag gac cca act ggg ctg atg ggc ggc att cat 288Asn Leu Val Glu Val Glu Asp Pro Thr Gly Leu Met Gly Gly Ile His
85 90 95cgc cgt tcg cct aaa aat ccg ctt atc gac gga ccc gcc agc ggt ttc 336Arg Arg Ser Pro Lys Asn Pro Leu Ile Asp Gly Pro Ala Ser Gly Phe
100 105 110aca ccc cat tac cgc gat ccc atg atc agc cct gat ggt gat ggt tgg 384Thr Pro His Tyr Arg Asp Pro Met Ile Ser Pro Asp Gly Asp Gly Trp
115 120 125aaa atg gtt ctt ggg gcc caa cgc gaa aac ctc acc ggt gca gcg gtt 432Lys Met Val Leu Gly Ala Gln Arg Glu Asn Leu Thr Gly Ala Ala Val
130 135 140cta tac cgc tcg aca gat ctt gaa aac tgg gaa ttc tcc ggt gaa at479Leu Tyr Arg Ser Thr Asp Leu Glu Asn Trp Glu Phe Ser Gly Glu145 150 155<210>14<211>159<212>PRT<213>Corynebacterium thermoaminogenes<400>14Tyr Tyr Gln His Asp Pro Gly Phe Pro Phe Ala Pro Lys Arg Thr Gly 1 5 10 15Trp Ala His Thr Thr Thr Pro Leu Thr Gly Pro Gln Arg Leu Gln Trp
20 25 30Thr His Leu Pro Asp Ala Leu Tyr Pro Asp Ala Ser Tyr Asp Leu Asp
35 40 45Gly Cys Tyr Ser Gly Gly Ala Val Phe Thr Asp Gly Thr Leu Lys Leu
50 55 60Phe Tyr Thr Gly Asn Leu Lys Ile Asp Gly Lys Arg Arg Ala Thr Gln 65 70 75 80Asn Leu Val Glu Val Glu Asp Pro Thr Gly Leu Met Gly Gly Ile His
85 90 95Arg Arg Ser Pro Lys Asn Pro Leu Ile Asp Gly Pro Ala Ser Gly Phe
100 105 110Thr Pro His Tyr Arg Asp Pro Met Ile Ser Pro Asp Gly Asp Gly Trp
115 120 125Lys Met Val Leu Gly Ala Gln Arg Glu Asn Leu Thr Gly Ala Ala Val
130 135 140Leu Tyr Arg Ser Thr Asp Leu Glu Asn Trp Glu Phe Ser Gly Glu145 150 155<210>15<211>490<212>DNA<213>Corynebacterium thermoaminogenes<400>15attttaatgg atattatcta tattttatca atattatcct tatgcacctg aatggggacc 60aatgcattgg ggacacgcac gtagtaaaga tttagttcat tgggaaacat taccgattgc 120tttagaacct ggagatgaag aagaaaaatg gttgtttctc tggtacaggt atagtcaaag 180atgataagtt gtatttattt tatacaggtc accattatta taatgacgat gatcccgatc 240atttttggca aaatcaaaat atggcttata gtgaagatgg cattcatttt caaaaatata 300aacaaaatgc aatcattcct accccacctg aagataatac acatcacttc agagatccaa 360aggtatggga acatccatgg cttattatta catgatagta ggtagtcaaa atgatagaga 420attaggacgt attatcttat atcgttctga ggatttatag aggggaattc tggtcctgag 480atcaatccaa490<210>16<211>4254<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(637)..(1362)<220><221>CDS<222>(1434)..(2315)<220><221>CDS<222>(2432)..(3115)<220><221>CDS<222>(3235)..(4065)<400>16tcacggcgcg cagattaccc agtgtgccgt agagacgctg atcggcattc tcacgcaccg 60cgcaggtgtt gaagacgatg agatcagggg tgtcaccctc ccccgccgcg gtgtaaccgg 120cctcctcgag cagaccggag agacgctcgg aatcgtggac gttcatctgg cagccgaagg 180tacgcacctc ataggtgcgg gcagtggtgc cctcccggtt cccccgcgcc gggagggtgt 240cggcggggtg gtccgggtgg gatggatggg tgttcatctg gtgggtatca atctgctgcg 300tcacgggagg taattgtatc ggccgcgggc accctgacat aaacgtccga tccagaggaa 360cgcaaccccg tggagtgtcg cagccatgca ggttgggcaa caccgtaacg gaacctagca 420gagtggtagg attgacttca cattctttac ctattgagct attgataaaa tccgggcgga 480aatggaaatc acccccacaa atcaccccaa ctgacctgtg gaaagggcga gaaatccagg 540gaaattcatt tcaaaatgga ctcaatcaca ggatttaccc cacatgaccc aacattcctt 600tatgctatcc ccatgacgca gaccacaaat cacccg atg atc aag atg acg ggg 654
Met Ile Lys Met Thr Gly
1 5gtg cag aag ttc ttc gat gac ttc cag gcc ctg acc gat atc aat ctt 702Val Gln Lys Phe Phe Asp Asp Phe Gln Ala Leu Thr Asp Ile Asn Leu
10 15 20gag gtc ccc gcg gga cag gtc gtt gtt gtt ctc ggc ccg tcc ggt tcc 750Glu Val Pro Ala Gly Gln Val Val Val Val Leu Gly Pro Ser Gly Ser
25 30 35gga aag tcg acg ctg tgc cgc acc atc aac cgc ctc gaa acc atc gag 798Gly Lys Ser Thr Leu Cys Arg Thr Ile Asn Arg Leu Glu Thr Ile Glu
40 45 50gag gga acc atc gag atc gat gga aaa ctg ctt ccg gag gag ggc aag 846Glu Gly Thr Ile Glu Ile Asp Gly Lys Leu Leu Pro Glu Glu Gly Lys 55 60 65 70gac ctg gcc aag atc cgt gcc gac gtg ggc atg gtg ttc cag tct ttc 894Asp Leu Ala Lys Ile Arg Ala Asp Val Gly Met Val Phe Gln Ser Phe
75 80 85aac ctc ttc ccc cac ctc acc atc aag gac aat gtc acc ctc ggc ccg 942Asn Leu Phe Pro His Leu Thr Ile Lys Asp Asn Val Thr Leu Gly Pro
90 95 100atg aag gtc cgg aag atg aag aag tcc gag gcc aat gag gtg gcc atg 990Met Lys Val Arg Lys Met Lys Lys Ser Glu Ala Asn Glu Val Ala Met
105 110 115aag ctg ttg gaa cgc gtc ggc atc gcc aac cag gcc gag aaa tac ccg 1038Lys Leu Leu Glu Arg Val Gly Ile Ala Asn Gln Ala Glu Lys Tyr Pro
120 125 130gca cag ctc tcg ggc ggg cag cag cag cgc gtg gcc atc gcc cgc gca 1086Ala Gln Leu Ser Gly Gly Gln Gln Gln Arg Val Ala Ile Ala Arg Ala135 140 145 150ctg gcg atg aac ccc aag atc atg ctt ttc gac gaa cca acc tcc gcc 1134Leu Ala Met Asn Pro Lys Ile Met Leu Phe Asp Glu Pro Thr Ser Ala
155 160 165ctc gac ccc gag atg gtc aac gag gtt ctg gac gtc atg gcg agt ctg 1182Leu Asp Pro Glu Met Val Asn Glu Val Leu Asp Val Met Ala Ser Leu
170 175 180gcc aag gaa ggc atg acc atg gtg tgt gtc acc cac gag atg ggt ttc 1230Ala Lys Glu Gly Met Thr Met Val Cys Val Thr His Glu Met Gly Phe
185 190 195gca cgc agg gcc gca gac cgt gtg ctg ttc atg tct gac ggc gcc atc 1278Ala Arg Arg Ala Ala Asp Arg Val Leu Phe Met Ser Asp Gly Ala Ile
200 205 210gtc gag gac tcc gac ccg gag acc ttc ttc acc aat cca caa acc gac 1326Val Glu Asp Ser Asp Pro Glu Thr Phe Phe Thr Asn Pro Gln Thr Asp215 220 225 230cgg gcg aag gat ttc ctg ggc aag atc ctc gcc cac tgacctcccc1372Arg Ala Lys Asp Phe Leu Gly Lys Ile Leu Ala His
235 240tcactctgtg tccaactccc ccgctggcca aaatcagcga ccatgaccaa caggagcatc 1432a atg tcg cac aaa cgc atg ttc acc cgt ctc gcc gca gcc acc agc gca 1481 Met Ser His Lys Arg Met Phe Thr Arg Leu Ala Ala Ala Thr Ser Ala
245 250 255gct gtt ctc gcc ggc atc acc ctc acc gcc tgt ggt gat tcc gag ggt 1529Ala Val Leu Ala Gly Ile Thr Leu Thr Ala Cys Gly Asp Ser Glu Gly
260 265 270ggt gac ggt ctg ctc gcc gcc atc gaa aat ggc aat gtc acc atc ggc 1577Gly Asp Gly Leu Leu Ala Ala Ile Glu Asn Gly Asn Val Thr Ile Gly275 280 285 290acc aag tac gat cag ccg ggt ctg gga ctg cgt aac ccg gac aat tcc 1625Thr Lys Tyr Asp Gln Pro Gly Leu Gly Leu Arg Asn Pro Asp Asn Ser
295 300 305atg agc gga ctg gat gtc gac gtc gcg cag tac gtg gtc aac tcc atc 1673Met Ser Gly Leu Asp Val Asp Val Ala Gln Tyr Val Val Asn Ser Ile
310 315 320gcc gat gac aac ggt tgg gat cac ccc acc gtg gaa tgg cgc gag acc 1721Ala Asp Asp Asn Gly Trp Asp His Pro Thr Val Glu Trp Arg Glu Thr
325 330 335ccc tcc gcc cag cgc gag acc ctc atc cag aac ggt gag gtg gat atg 1769Pro Ser Ala Gln Arg Glu Thr Leu Ile Gln Asn Gly Glu Val Asp Met
340 345 350atc gcc gca acc tac tcc atc aac ccc gga cgc tcc gaa tcg gtg aac 1817Ile Ala Ala Thr Tyr Ser Ile Asn Pro Gly Arg Ser Glu Ser Val Asn355 360 365 370ttc ggt gga cca tac ctc ctc acc cac cag gcc ctc ctg gtc cgc gag 1865Phe Gly Gly Pro Tyr Leu Leu Thr His Gln Ala Leu Leu Val Arg Glu
375 380 385gac gat gac cgc atc cag acc ctc gag gac ctc gat gac ggc ctg atc 1913Asp Asp Asp Arg Ile Gln Thr Leu Glu Asp Leu Asp Asp Gly Leu Ile
390 395 400ctg tgt tcc gtt acc gga tcc acc ccc gcc cag aag gtc aag gat gtc 1961Leu Cys Ser Val Thr Gly Ser Thr Pro Ala Gln Lys Val Lys Asp Val
405 410 415ctc ccc ggc gtc cag ctg cag gaa tac gac acc tac tcc tcc tgt gtg 2009Leu Pro Gly Val Gln Leu Gln Glu Tyr Asp Thr Tyr Ser Ser Cys Val
420 425 430gag gca ctg agc cag ggc aac gtc gat gca atg acc acc gac gcc acc 2057Glu Ala Leu Ser Gln Gly Asn Val Asp Ala Met Thr Thr Asp Ala Thr435 440 445 450atc ctc ttc ggc tac gcg cag cag cgc gaa ggt gaa ttc cgc gtc gtg 2105Ile Leu Phe Gly Tyr Ala Gln Gln Arg Glu Gly Glu Phe Arg Val Val
455 460 465gag atg gaa cag gac ggc gag ccg ttc acc aat gag tac tac ggc atc 2153Glu Met Glu Gln Asp Gly Glu Pro Phe Thr Asn Glu Tyr Tyr Gly Ile
470 475 480ggt atc acc aag gat gac acc gaa gcc acc gat gcg atc aac gca gcg 2201Gly Ile Thr Lys Asp Asp Thr Glu Ala Thr Asp Ala Ile Asn Ala Ala
485 490 495ttg gag cgt atg tac gcc gac ggt tcc ttc cag cgt ttc ctc acc gag 2249Leu Glu Arg Met Tyr Ala Asp Gly Ser Phe Gln Arg Phe Leu Thr Glu
500 505 510aac ctc ggc gag gat tcc cag gtt gtc cag gag ggc acc ccg ggt gac 2297Asn Leu Gly Glu Asp Ser Gln Val Val Gln Glu Gly Thr Pro Gly Asp515 520 525 530ctc tcc ttc ctg gac gag tgacctgacg gggccgaacg cccgatgagc 2345Leu Ser Phe Leu Asp Glu
535atgcgtggcc cccgcatccc ggggtgccac gcatcatcac tttcaccact gatcccctac 2405cgttccttac cgaggagaaa ttcccc atg agt aca tta tgg gcg gat ctg ggt 2458
Met Ser Thr Leu Trp Ala Asp Leu Gly
540 545ccg tca ctc cta ccc gca ttc tgg gtg aca atc caa ctc acc gtc tat 2506Pro Ser Leu Leu Pro Ala Phe Trp Val Thr Ile Gln Leu Thr Val Tyr
550 555 560tcc gcc atc gga tcc atg atc ctc ggt acc atc ctc acc gcc atg agg 2554Ser Ala Ile Gly Ser Met Ile Leu Gly Thr Ile Leu Thr Ala Met Arg
565 570 575gtg tcc ccg gtg aag atc ctg cgc agc ata tcc acc gcc tac atc aac 2602Val Ser Pro Val Lys Ile Leu Arg Ser Ile Ser Thr Ala Tyr Ile Asn
580 585 590acg gtc cgt aac acc cca ctg acc ctg gtg atc ctg ttc tgt tcc ttc 2650Thr Val Arg Asn Thr Pro Leu Thr Leu Val Ile Leu Phe Cys Ser Phe
595 600 605ggc ctg tat cag aat ctc ggt ctc acc ctc gcc ggt cgc gac agt tcc 2698Gly Leu Tyr Gln Asn Leu Gly Leu Thr Leu Ala Gly Arg Asp Ser Ser610 615 620 625acc ttt ctg gcc gat aac aac ttc cgg ctc gcg gtg ctc gga ttc atc 2746Thr Phe Leu Ala Asp Asn Asn Phe Arg Leu Ala Val Leu Gly Phe Ile
630 635 640ctg tac acc tcc gcc ttc gtt gcg gaa tca ctc cgg tca ggc atc aac 2794Leu Tyr Thr Ser Ala Phe Val Ala Glu Ser Leu Arg Ser Gly Ile Asn
645 650 655acc gtg cac ttc ggg cag gcg gag gcc gcc cgg tcg ctg gga ctc ggt 2842Thr Val His Phe Gly Gln Ala Glu Ala Ala Arg Ser Leu Gly Leu Gly
660 665 670ttc agt gac atc ttc cgg tcc atc atc ttc ccc cag gcg gtg cgt gcc 2890Phe Ser Asp Ile Phe Arg Ser Ile Ile Phe Pro Gln Ala Val Arg Ala
675 680 685gcc atc atc ccg ctg ggc aac acc ctc atc gcc ctg acc aag aac acc 2938Ala Ile Ile Pro Leu Gly Asn Thr Leu Ile Ala Leu Thr Lys Asn Thr690 695 700 705acg atc gcg tcc gtg atc ggc gtc ggt gag gcc tcg ctg ctg atg aag 2986Thr Ile Ala Ser Val Ile Gly Val Gly Glu Ala Ser Leu Leu Met Lys
710 715 720tcc acg att gaa aat cat gcc aac atg ctc ttc gtc gtg ttc gcc atc 3034Ser Thr Ile Glu Asn His Ala Asn Met Leu Phe Val Val Phe Ala Ile
725 730 735ttc gcc gtc ggc ttc atg atc ctc acc ctc ccc atg ggc ctg ggg ctt 3082Phe Ala Val Gly Phe Met Ile Leu Thr Leu Pro Met Gly Leu Gly Leu
740 745 750gga aaa ctc gct gag aaa atg gcg gtg aag aaa taatgtcctc ctccgtacgc 3135Gly Lys Leu Ala Glu Lys Met Ala Val Lys Lys
755 760gcaacagtcc tctacgacgc ccccggcccc cggggacgca ggtccaacac catcatcacc 3195atcgccacca ccctggtggc agtggccgtc ctgttctgg gtg ggc agt gtt ctc3249
Val Gly Ser Val Leu
765cag gaa aac ggc cag ttg gac ggc gac aaa tgg acc ccg ttc ctc gat 3297Gln Glu Asn Gly Gln Leu Asp Gly Asp Lys Trp Thr Pro Phe Leu Asp770 775 780 785ccc cag acc tgg acc acc tat ctt ctg ccc ggc ctg tgg gga acc ctg 3345Pro Gln Thr Trp Thr Thr Tyr Leu Leu Pro Gly Leu Trp Gly Thr Leu
790 795 800aag gca gcg gtg gcc tcc atc ctt ctc gcg ctg atc atg ggc acc ctg 3393Lys Ala Ala Val Ala Ser Ile Leu Leu Ala Leu Ile Met Gly Thr Leu
805 810 815ctc ggg ctc gga cgc atc tcc gaa atc cgg ctc ctg cgc tgg ttc tgc 3441Leu Gly Leu Gly Arg Ile Ser Glu Ile Arg Leu Leu Arg Trp Phe Cys
820 825 830ggg atc atc atc gag acc ttc cgt gcc atc ccg gtg ctg atc ctc atg 3489Gly Ile Ile Ile Glu Thr Phe Arg Ala Ile Pro Val Leu Ile Leu Met
835 840 845atc ttc gcc tat cag ttg ttc gcc cgt tac cag ctc gtt cca tca cgc 3537Ile Phe Ala Tyr Gln Leu Phe Ala Arg Tyr Gln Leu Val Pro Ser Arg850 855 860 865cag ctg gcc ttc gcc gcg gtg gtc ttc ggt ctc acc atg tac aac ggc 3585Gln Leu Ala Phe Ala Ala Val Val Phe Gly Leu Thr Met Tyr Asn Gly
870 875 880tcc gtc atc gcc gag atc ctt aga tcg ggt atc gcc tcc ctg ccg aag 3633Ser Val Ile Ala Glu Ile Leu Arg Ser Gly Ile Ala Ser Leu Pro Lys
885 890 895gga cag cgt gag gcg gcg atc gcc ctg ggc atg tca acc cgc cag acc 3681Gly Gln Arg Glu Ala Ala Ile Ala Leu Gly Met Ser Thr Arg Gln Thr
900 905 910acc tgg tcg atc ctg ctc ccc cag gcg gtg gca gcg atg ctg ccc gcc 3729Thr Trp Ser Ile Leu Leu Pro Gln Ala Val Ala Ala Met Leu Pro Ala
915920 925ctg atc gcg cag atg gtc atc gcg ctg aag gac tcc gcc ctc ggt tac 3777Leu Ile Ala Gln Met Val Ile Ala Leu Lys Asp Ser Ala Leu Gly Tyr930 935 940 945cag atc ggt tat atc gag gtg gta cgc tcc ggt atc cag tcc gca tcc 3825Gln Ile Gly Tyr Ile Glu Val Val Arg Ser Gly Ile Gln Ser Ala Ser
950 955 960gtc aac cgg aac tac ctg gct gcc ctc gcg gtg gtc gcg gtc atc atg 3873Val Asn Arg Asn Tyr Leu Ala Ala Leu Ala Val Val Ala Val Ile Met
965 970 975atc ctg atc aac ttc gca ctg acc gca ctg gca gag cgt atc cag cgt 3921Ile Leu Ile Asn Phe Ala Leu Thr Ala Leu Ala Glu Arg Ile Gln Arg
980 985 990cag ctg cgt gcc gga cgt gcc cgc agg aac att gtg gca aag gtg ccc 3969Gln Leu Arg Ala Gly Arg Ala Arg Arg Asn Ile Val Ala Lys Val Pro
99510001005gag gaa ccc gat cag ggc ctg gat acc aag gac aat gtg aac gtg gat 4017Glu Glu Pro Asp Gln Gly Leu Asp Thr Lys Asp Asn Val Asn Val Asp1010 101510201025tgg cac gat ccc gat tac aag gaa gtc aaa cac ccg gga ccg tca ttc 4065Trp His Asp Pro Asp Tyr Lys Glu Val Lys His Pro Gly Pro Ser Phe
103010351040tgacaggtcc ctggatcccc gctgcggtca ggaggcgggt gcaacaatga agtccggctg 4125cccagatgtc tggggcagcc ggactttgtg gcagatcaat gctgactgag gtcctcgatg 4185cgctcatcga gagcctcccg ggccaggtcc atcgacatac ccgcggggaa tccacgacgg 4245gcaagtgct 4254<210>17<211>242<212>PRT<213>Corynebacterium thermoaminogenes<400>17Met Ile Lys Met Thr Gly Val Gln Lys Phe Phe Asp Asp Phe Gln Ala 1 5 10 15Leu Thr Asp Ile Asn Leu Glu Val Pro Ala Gly Gln Val Val Val Val
20 25 30Leu Gly Pro Ser Gly Ser Gly Lys Ser Thr Leu Cys Arg Thr Ile Asn
35 40 45Arg Leu Glu Thr Ile Glu Glu Gly Thr Ile Glu Ile Asp Gly Lys Leu
50 55 60Leu Pro Glu Glu Gly Lys Asp Leu Ala Lys Ile Arg Ala Asp Val Gly 65 70 75 80Met Val Phe Gln Ser Phe Asn Leu Phe Pro His Leu Thr Ile Lys Asp
85 90 95Asn Val Thr Leu Gly Pro Met Lys Val Arg Lys Met Lys Lys Ser Glu
100 105 110Ala Asn Glu Val Ala Met Lys Leu Leu Glu Arg Val Gly Ile Ala Asn
115 120 125Gln Ala Glu Lys Tyr Pro Ala Gln Leu Ser Gly Gly Gln Gln Gln Arg
130 135 140Val Ala Ile Ala Arg Ala Leu Ala Met Asn Pro Lys Ile Met Leu Phe145 150 155 160Asp Glu Pro Thr Ser Ala Leu Asp Pro Glu Met Val Asn Glu Val Leu
165 170 175Asp Val Met Ala Ser Leu Ala Lys Glu Gly Met Thr Met Val Cys Val
180 185 190Thr His Glu Met Gly Phe Ala Arg Arg Ala Ala Asp Arg Val Leu Phe
195 200 205Met Ser Asp Gly Ala Ile Val Glu Asp Ser Asp Pro Glu Thr Phe Phe
210 215 220Thr Asn Pro Gln Thr Asp Arg Ala Lys Asp Phe Leu Gly Lys Ile Leu225 230 235 240Ala His<210>18<211>294<212>PRT<213>Corynebacterium thermoaminogenes<400>18Met Ser His Lys Arg Met Phe Thr Arg Leu Ala Ala Ala Thr Ser Ala 1 5 10 15Ala Val Leu Ala Gly Ile Thr Leu Thr Ala Cys Gly Asp Ser Glu Gly
20 25 30Gly Asp Gly Leu Leu Ala Ala Ile Glu Asn Gly Asn Val Thr Ile Gly
35 40 45Thr Lys Tyr Asp Gln Pro Gly Leu Gly Leu Arg Asn Pro Asp Asn Ser
50 55 60Met Ser Gly Leu Asp Val Asp Val Ala Gln Tyr Val Val Asn Ser Ile 65 70 75 80Ala Asp Asp Asn Gly Trp Asp His Pro Thr Val Glu Trp Arg Glu Thr
85 90 95Pro Ser Ala Gln Arg Glu Thr Leu Ile Gln Asn Gly Glu Val Asp Met
100 105 110Ile Ala Ala Thr Tyr Ser Ile Asn Pro Gly Arg Ser Glu Ser Val Asn
115 120 125Phe Gly Gly Pro Tyr Leu Leu Thr His Gln Ala Leu Leu Val Arg Glu
130 135 140Asp Asp Asp Arg Ile Gln Thr Leu Glu Asp Leu Asp Asp Gly Leu Ile145 150 155 160Leu Cys Ser Val Thr Gly Ser Thr Pro Ala Gln Lys Val Lys Asp Val
165 170 175Leu Pro Gly Val Gln Leu Gln Glu Tyr Asp Thr Tyr Ser Ser Cys Val
180 185 190Glu Ala Leu Ser Gln Gly Asn Val Asp Ala Met Thr Thr Asp Ala Thr
195 200 205Ile Leu Phe Gly Tyr Ala Gln Gln Arg Glu Gly Glu Phe Arg Val Val
210 215 220Glu Met Glu Gln Asp Gly Glu Pro Phe Thr Asn Glu Tyr Tyr Gly Ile225 230 235 240Gly Ile Thr Lys Asp Asp Thr Glu Ala Thr Asp Ala Ile Asn Ala Ala
245 250 255Leu Glu Arg Met Tyr Ala Asp Gly Ser Phe Gln Arg Phe Leu Thr Glu
260 265 270Asn Leu Gly Glu Asp Ser Gln Val Val Gln Glu Gly Thr Pro Gly Asp
275 280 285Leu Ser Phe Leu Asp Glu
290<210>19<211>228<212>PRT<213>Corynebacterium thermoaminogenes<400>19Met Ser Thr Leu Trp Ala Asp Leu Gly Pro Ser Leu Leu Pro Ala Phe 1 5 l0 15Trp Val Thr Ile Gln Leu Thr Val Tyr Ser Ala Ile Gly Ser Met Ile
20 25 30Leu Gly Thr Ile Leu Thr Ala Met Arg Val Ser Pro Val Lys Ile Leu
35 40 45Arg Ser Ile Ser Thr Ala Tyr Ile Asn Thr Val Arg Asn Thr Pro Leu
50 55 60Thr Leu Val Ile Leu Phe Cys Ser Phe Gly Leu Tyr Gln Asn Leu Gly 65 70 75 80Leu Thr Leu Ala Gly Arg Asp Ser Ser Thr Phe Leu Ala Asp Asn Asn
85 90 95Phe Arg Leu Ala Val Leu Gly Phe Ile Leu Tyr Thr Ser Ala Phe Val
100 105 110Ala Glu Ser Leu Arg Ser Gly Ile Asn Thr Val His Phe Gly Gln Ala
115120 125Glu Ala Ala Arg Ser Leu Gly Leu Gly Phe Ser Asp Ile Phe Arg Ser
130 135 140Ile Ile Phe Pro Gln Ala Val Arg Ala Ala Ile Ile Pro Leu Gly Asn145 150 155 160Thr Leu Ile Ala Leu Thr Lys Asn Thr Thr Ile Ala Ser Val Ile Gly
165 170 175Val Gly Glu Ala Ser Leu Leu Met Lys Ser Thr Ile Glu Asn His Ala
180 185 190Asn Met Leu Phe Val Val Phe Ala Ile Phe Ala Val Gly Phe Met Ile
195 200 205Leu Thr Leu Pro Met Gly Leu Gly Leu Gly Lys Leu Ala Glu Lys Met
210 215 220Ala Val Lys Lys225<210>20<211>277<212>PRT<213>Corynebacterium thermoaminogenes<400>20Val Gly Ser Val Leu Gln Glu Asn Gly Gln Leu Asp Gly Asp Lys Trp 1 5 10 15Thr Pro Phe Leu Asp Pro Gln Thr Trp Thr Thr Tyr Leu Leu Pro Gly
20 25 30Leu Trp Gly Thr Leu Lys Ala Ala Val Ala Ser Ile Leu Leu Ala Leu
35 40 45Ile Met Gly Thr Leu Leu Gly Leu Gly Arg Ile Ser Glu Ile Arg Leu
50 55 60Leu Arg Trp Phe Cys Gly Ile Ile Ile Glu Thr Phe Arg Ala Ile Pro 65 70 75 80Val Leu Ile Leu Met Ile Phe Ala Tyr Gln Leu Phe Ala Arg Tyr Gln
85 90 95Leu Val Pro Ser Arg Gln Leu Ala Phe Ala Ala Val Val Phe Gly Leu
100 105 110Thr Met Tyr Asn Gly Ser Val Ile Ala Glu Ile Leu Arg Ser Gly Ile
115 120 125Ala Ser Leu Pro Lys Gly Gln Arg Glu Ala Ala Ile Ala Leu Gly Met
130 135 140Ser Thr Arg Gln Thr Thr Trp Ser Ile Leu Leu Pro Gln Ala Val Ala145 150 155 160Ala Met Leu Pro Ala Leu Ile Ala Gln Met Val Ile Ala Leu Lys Asp
165 170 175Ser Ala Leu Gly Tyr Gln Ile Gly Tyr Ile Glu Val Val Arg Ser Gly
180 185 190Ile Gln Ser Ala Ser Val Asn Arg Asn Tyr Leu Ala Ala Leu Ala Val
195 200 205Val Ala Val Ile Met Ile Leu Ile Asn Phe Ala Leu Thr Ala Leu Ala
210 215 220Glu Arg Ile Gln Arg Gln Leu Arg Ala Gly Arg Ala Arg Arg Asn Ile225 230 235 240Val Ala Lys Val Pro Glu Glu Pro Asp Gln Gly Leu Asp Thr Lys Asp
245 250 255Asn Val Asn Val Asp Trp His Asp Pro Asp Tyr Lys Glu Val Lys His
260 265 270Pro Gly Pro Ser Phe
275<210>21<211>3598<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(454)..(3222)<400>21agcacggcca aacatgagag aaacttcaca ttttgaattt cccctttcct gcatatggaa 60aaccgccggt gacacccctg ccatttgggc agctcccccc acctcaccat gtccacattt 120tccataatgt ggcctgtaac acccttgggc tcaaggcttc cacgccccac cgggaccctc 180atcagcaggt gaaacagacc ctcctgcaat gctttgttaa aaagaaccgc cctttgtgcg 240tatccttgtg tcaattgtgc gcgcactgcc accagctttc ctcaggattg aacacggtcg 300ggaaatcctc cccggatacc ctgcacgccc cacctcccac accgacaccg gcggggaggg 360ccgggcacgt tttcagctgc gggtgatgga agcggtcgcc ggtcccccgg tcgcataaac 420gaaatgaaaa acattccaac aggaggtgtg gaa atg gcc gat caa gca aaa ctt 474
Met Ala Asp Gln Ala Lys Leu
1 5ggt ggc aaa ccc aca gat gac acc aac ttc gcg atg atc cgt gat ggc 522Gly Gly Lys Pro Thr Asp Asp Thr Asn Phe Ala Met Ile Arg Asp Gly
10 15 20gtt gca tct tat ttg aac gac tcc gac ccg gag gag acc aag gag tgg 570Val Ala Ser Tyr Leu Asn Asp Ser Asp Pro Glu Glu Thr Lys Glu Trp
25 30 35atg gac tcc cta gac ggt cta ctg cag gat tcc tct ccg gag cgc gcc 618Met Asp Ser Leu Asp Gly Leu Leu Gln Asp Ser Ser Pro Glu Arg Ala 40 45 50 55cgt tac ctg atg ctg cgc ctg ctg gag cgg gca tcc gcc aag cgt gtc 666Arg Tyr Leu Met Leu Arg Leu Leu Glu Arg Ala Ser Ala Lys Arg Val
60 65 70cca ctg ccc ccg atg acg tcc acc gat tac gtc aac acc atc ccc aca 714Pro Leu Pro Pro Met Thr Ser Thr Asp Tyr Val Asn Thr Ile Pro Thr
75 80 85tcc atg gag ccc gat ttc ccg ggt gat gag gag atg gag aag cgc tac 762Ser Met Glu Pro Asp Phe Pro Gly Asp Glu Glu Met Glu Lys Arg Tyr
90 95 100cgc cgc tgg atg cgc tgg aac gcc gcc atc atg gtg cac cgt gcc cag 810Arg Arg Trp Met Arg Trp Asn Ala Ala Ile Met Val His Arg Ala Gln
105 110 115cgc ccg gga atc ggt gtg ggt ggg cac atc tcc acc tac gcc ggc gcc 858Arg Pro Gly Ile Gly Val Gly Gly His Ile Ser Thr Tyr Ala Gly Ala120 125 130 135gcc cca ctc tac gag gtc ggt ttc aac cac ttc ttc cgc ggc aag gac 906Ala Pro Leu Tyr Glu Val Gly Phe Asn His Phe Phe Arg Gly Lys Asp
140 145 150cac ccg ggt ggc ggt gac cag gtc ttc ttc cag ggt cac gcc tcc ccg 954His Pro Gly Gly Gly Asp Gln Val Phe Phe Gln Gly His Ala Ser Pro
155 160 165ggc atg tac gcc cgc gcc ttc ctc gag ggc cgt ctc acc gag agc gat 1002Gly Met Tyr Ala Arg Ala Phe Leu Glu Gly Arg Leu Thr Glu Ser Asp
170 175 180ctg gac agc ttc cgc cag gag gtc tcc tac gaa ggt ggt ggc atc ccg 1050Leu Asp Ser Phe Arg Gln Glu Val Ser Tyr Glu Gly Gly Gly Ile Pro
185 190 195tcc tac ccg cac ccg cac ggc atg ccg gac ttc tgg gag ttc ccg acc 1098Ser Tyr Pro His Pro His Gly Met Pro Asp Phe Trp Glu Phe Pro Thr200 205 210 215gtg tcc atg ggc ctc ggg ccc atg gat gcc atc tac cag gcg cgc ttc 1146Val Ser Met Gly Leu Gly Pro Met Asp Ala Ile Tyr Gln Ala Arg Phe
220 225 230aac cgc tac ctg cac aac cgt ggc atc aag gac acc tcg gag cag cac 1194Asn Arg Tyr Leu His Asn Arg Gly Ile Lys Asp Thr Ser Glu Gln His
235 240 245gtc tgg gca ttc ctc ggt gac ggc gag atg gat gag ccg gag tcc cgt 1242Val Trp Ala Phe Leu Gly Asp Gly Glu Met Asp Glu Pro Glu Ser Arg
250 255 260ggt ctc atc cac cag gct gcg ctg aac aac ctg gac aac ctc acc ttc 1290Gly Leu Ile His Gln Ala Ala Leu Asn Asn Leu Asp Asn Leu Thr Phe
265 270 275gtg atc aac tgc aac ctg cag cgt ctt gat ggc ccg gtc cgc ggt aac 1338Val Ile Asn Cys Asn Leu Gln Arg Leu Asp Gly Pro Val Arg Gly Asn280 285 290 295acc aag atc atc cag gaa ctc gag tcc ttc ttc cgt ggt gcc ggc tgg 1386Thr Lys Ile Ile Gln Glu Leu Glu Ser Phe Phe Arg Gly Ala Gly Trp
300 305 310tcc gtc atc aag gtc atc tgg ggc cgt gag tgg gat gaa ctg ctg gag 1434Ser Val Ile Lys Val Ile Trp Gly Arg Glu Trp Asp Glu Leu Leu Glu
315 320 325aag gac cag gac ggt gct ctt gtc gag gtc atg aac aac acc tcc gac 1482Lys Asp Gln Asp Gly Ala Leu Val Glu Val Met Asn Asn Thr Ser Asp
330 335 340ggt gac tac cag acc ttc aag gcc aat gac ggt gcc tac gtc cgt gag 1530Gly Asp Tyr Gln Thr Phe Lys Ala Asn Asp Gly Ala Tyr Val Arg Glu
345 350 355cac ttc ttc ggc cgt gac ccc cgc acc ctc aag ctc gtc gag gac atg 1578His Phe Phe Gly Arg Asp Pro Arg Thr Leu Lys Leu Val Glu Asp Met360 365 370 375acc gac gag gag atc tgg aag ctg ccc cgt ggt ggc cat gac tac cgt 1626Thr Asp Glu Glu Ile Trp Lys Leu Pro Arg Gly Gly His Asp Tyr Arg
380 385 390aag gtc tac gcc gcc tac aag cgt gcg ctg gag acc aag gac cgc ccg 1674Lys Val Tyr Ala Ala Tyr Lys Arg Ala Leu Glu Thr Lys Asp Arg Pro
395 400 405acc gtc att ctc gcc cat acc atc aag ggc tac ggc ctg ggc cac aac 1722Thr Val Ile Leu Ala His Thr Ile Lys Gly Tyr Gly Leu Gly His Asn
410 415 420ttc gag ggc cgc aac gcg acc cac cag atg aag aag ctg acc ctg gat 1770Phe Glu Gly Arg Asn Ala Thr His Gln Met Lys Lys Leu Thr Leu Asp
425 430 435gac ctg aag ctg ttc cgt gac aag cag ggt ctg ccc atc acc gat gag 1818Asp Leu Lys Leu Phe Arg Asp Lys Gln Gly Leu Pro Ile Thr Asp Glu440 445 450 455gag ctg gag aag gat ccc tac ctg cct ccg tac tac cac ccg ggt gag 1866Glu Leu Glu Lys Asp Pro Tyr Leu Pro Pro Tyr Tyr His Pro Gly Glu
460 465 470gac gca ccg gag atc aag tac atg aag gag cgt cgc cag gcg ctc ggt 1914Asp Ala Pro Glu Ile Lys Tyr Met Lys Glu Arg Arg Gln Ala Leu Gly
475 480 485ggt ttc ctg ccg gag cgc cgt gag aag tac gag cca ctg cag gtt ccc 1962Gly Phe Leu Pro Glu Arg Arg Glu Lys Tyr Glu Pro Leu Gln Val Pro
490 495 500ccg ctg gac aag ctg cgg tcc gtg cgc aag ggt tcc ggc aag cag cag 2010Pro Leu Asp Lys Leu Arg Ser Val Arg Lys Gly Ser Gly Lys Gln Gln
505 510 515gtg gcc acc acc atg gcc acg gtg cgt acc ttc aag gaa ctc atg cgg 2058Val Ala Thr Thr Met Ala Thr Val Arg Thr Phe Lys Glu Leu Met Arg520 525 530 535gac aag aac ctg gcc gac cgc ttg gtc ccg atc atc ccg gat gag gcc 2106Asp Lys Asn Leu Ala Asp Arg Leu Val Pro Ile Ile Pro Asp Glu Ala
540 545 550cgc acc ttc ggc ctg gac tcc tgg ttc ccg acc ctg aaa atc tac aac 2154Arg Thr Phe Gly Leu Asp Ser Trp Phe Pro Thr Leu Lys Ile Tyr Asn
555 560 565ccg cac ggt cag aac tac gtg ccg gtc gac cat gac ctc atg ctg tcc 2202Pro His Gly Gln Asn Tyr Val Pro Val Asp His Asp Leu Met Leu Ser
570 575 580tac cgt gag gcc aag gac ggc cag atc ctg cat gag ggc atc aac gag 2250Tyr Arg Glu Ala Lys Asp Gly Gln Ile Leu His Glu Gly Ile Asn Glu
585 590 595gcc ggt tcc gtg gca tcg ttt atc gcc gcc gga acc tcc tac gcc acc 2298Ala Gly Ser Val Ala Ser Phe Ile Ala Ala Gly Thr Ser Tyr Ala Thr600 605 610 615cat ggc gag gcc atg atc ccg ctg tac atc ttc tac tcg atg ttc ggc 2346His Gly Glu Ala Met Ile Pro Leu Tyr Ile Phe Tyr Ser Met Phe Gly
620 625 630ttc cag cgc acc ggt gac ggc atc tgg gcc gca gcc gac cag atg acg 2394Phe Gln Arg Thr Gly Asp Gly Ile Trp Ala Ala Ala Asp Gln Met Thr
635 640 645cgt ggt ttc ctc ctg ggc gcc acc gcc ggt cgc acc acc ctg acc ggt 2442Arg Gly Phe Leu Leu Gly Ala Thr Ala Gly Arg Thr Thr Leu Thr Gly
650 655 660gag ggc ctc cag cac atg gat ggc cac tcc ccg atc ctg gcc tcc acc 2490Glu Gly Leu Gln His Met Asp Gly His Ser Pro Ile Leu Ala Ser Thr
665 670 675aac ccc ggt gtg gag acc tat gac ccg gcg ttc tcc tac gag atc gcg 2538Asn Pro Gly Val Glu Thr Tyr Asp Pro Ala Phe Ser Tyr Glu Ile Ala680 685 690 695cac ctg gtc cac cgc ggc atc gac cgc atg tac gga ccg ggc aag ggt 2586His Leu Val His Arg Gly Ile Asp Arg Met Tyr Gly Pro Gly Lys Gly
700 705 710gag aat gtc atc tac tac ctc acc atc tac aac gag cca acc ccg cag 2634Glu Asn Val Ile Tyr Tyr Leu Thr Ile Tyr Asn Glu Pro Thr Pro Gln
715 720 725ccg gct gag cct gag gat ctg gac gtc gag ggc ctg cac aag ggc atc 2682Pro Ala Glu Pro Glu Asp Leu Asp Val Glu Gly Leu His Lys Gly Ile
730 735 740tac ctc tac gac aag gcc gcc gag ggt gag ggc cat gag gcc tcg atc 2730Tyr Leu Tyr Asp Lys Ala Ala Glu Gly Glu Gly His Glu Ala Ser Ile
745 750 755ctg gcc tcc ggc atc ggc atg cag tgg gca ctg cgc gcc cgt gac atc 2778Leu Ala Ser Gly Ile Gly Met Gln Trp Ala Leu Arg Ala Arg Asp Ile760 765 770 775ctc gcc gag gat tac ggc atc cgt gcc aac atc ttc tcc gcc acc tcg 2826Leu Ala Glu Asp Tyr Gly Ile Arg Ala Asn Ile Phe Ser Ala Thr Ser
780 785 790tgg gtg gag ctg gcc cgc gac ggt gcc cgc cgt aac ctg gag gcg ctg 2874Trp Val Glu Leu Ala Arg Asp Gly Ala Arg Arg Asn Leu Glu Ala Leu
795 800 805cgc aac ccg ggt gcg gat gtc ggt gag gca ttc gtg acc acc cag ctg 2922Arg Asn Pro Gly Ala Asp Val Gly Glu Ala Phe Val Thr Thr Gln Leu
810 815 820aag aag ggt tcc ggc ccc tac gtc gcg gtg tcc gac ttc gcg acc gac 2970Lys Lys Gly Ser Gly Pro Tyr Val Ala Val Ser Asp Phe Ala Thr Asp
825 830 835ctg ccg aac cag atc cgc gag tgg gtt ccc ggt gac tac atc gtc ctc 3018Leu Pro Asn Gln Ile Arg Glu Trp Val Pro Gly Asp Tyr Ile Val Leu840 845 850 855ggt gcc gac ggc ttc ggt ttc tcc gat acc cgt ccg gca gcc cgt cgt 3066Gly Ala Asp Gly Phe Gly Phe Ser Asp Thr Arg Pro Ala Ala Arg Arg
860 865 870tac ttc aac atc gac gcc gag tcc atc gtc gtg gcg gtc ctg cgc ggc 3114Tyr Phe Asn Ile Asp Ala Glu Ser Ile Val Val Ala Val Leu Arg Gly
875 880 885ctg gtc cgc gag ggt gtc atc gat gcc tcc gtg gcg gcg cac gcg gct 3162Leu Val Arg Glu Gly Val Ile Asp Ala Ser Val Ala Ala His Ala Ala
890 895 900gag aag tac aag ctg tcc gac ccg acg gca cca cag gtc gat ccg gac 3210Glu Lys Tyr Lys Leu Ser Asp Pro Thr Ala Pro Gln Val Asp Pro Asp
905 910 915gca ccg atc gag tagacctgct tgtcgacgaa aaacaccccc gccccctcac 3262Ala Pro Ile Glu920atgatgaggg gggcgggggt gtgctcgttt acggcgggta caggggggta tcagcccagc 3322atcgccttat cggagagcgt cgcgcccttg atcttggcga attcctgcag cagatcccgc 3382acggtgagct tctgcttcac ctctgcgctg gcctcataga cgatccgtcc ctcgtgcatc 3442atgatgaggc ggttacccag gcggatagcc tgttccatgt tgtgggtgac catgagggtg 3502gtcagtttgc cgtcctcgac gatcttctcg gtcagggtgg tgaccagttc ggctcgctgg 3562gggtccaggg cggcggtgtg ttcgtcgaga agcatg 3598<210>22<211>923<212>PRT<213>Corynebacterium thermoaminogenes<400>22Met Ala Asp Gln Ala Lys Leu Gly Gly Lys Pro Thr Asp Asp Thr Asn 1 5 10 15Phe Ala Met Ile Arg Asp Gly Val Ala Ser Tyr Leu Asn Asp Ser Asp
20 25 30Pro Glu Glu Thr Lys Glu Trp Met Asp Ser Leu Asp Gly Leu Leu Gln
35 40 45Asp Ser Ser Pro Glu Arg Ala Arg Tyr Leu Met Leu Arg Leu Leu Glu
50 55 60Arg Ala Ser Ala Lys Arg Val Pro Leu Pro Pro Met Thr Ser Thr Asp 65 70 75 80Tyr Val Asn Thr Ile Pro Thr Ser Met Glu Pro Asp Phe Pro Gly Asp
85 90 95Glu Glu Met Glu Lys Arg Tyr Arg Arg Trp Met Arg Trp Asn Ala Ala
100 105 110Ile Met Val His Arg Ala Gln Arg Pro Gly Ile Gly Val Gly Gly His
115 120 125Ile Ser Thr Tyr Ala Gly Ala Ala Pro Leu Tyr Glu Val Gly Phe Asn
130 135 140His Phe Phe Arg Gly Lys Asp His Pro Gly Gly Gly Asp Gln Val Phe145 150 155 160Phe Gln Gly His Ala Ser Pro Gly Met Tyr Ala Arg Ala Phe Leu Glu
165 170 175Gly Arg Leu Thr Glu Ser Asp Leu Asp Ser Phe Arg Gln Glu Val Ser
180 185 190Tyr Glu Gly Gly Gly Ile Pro Ser Tyr Pro His Pro His Gly Met Pro
195 200 205Asp Phe Trp Glu Phe Pro Thr Val Ser Met Gly Leu Gly Pro Met Asp
210 215 220Ala Ile Tyr Gln Ala Arg Phe Asn Arg Tyr Leu His Asn Arg Gly Ile225 230 235 240Lys Asp Thr Ser Glu Gln His Val Trp Ala Phe Leu Gly Asp Gly Glu
245 250 255Met Asp Glu Pro Glu Ser Arg Gly Leu Ile His Gln Ala Ala Leu Asn
260 265 270Asn Leu Asp Asn Leu Thr Phe Val Ile Asn Cys Asn Leu Gln Arg Leu
275 280 285Asp Gly Pro Val Arg Gly Asn Thr Lys Ile Ile Gln Glu Leu Glu Ser
290 295 300Phe Phe Arg Gly Ala Gly Trp Ser Val Ile Lys Val Ile Trp Gly Arg305 310 315 320Glu Trp Asp Glu Leu Leu Glu Lys Asp Gln Asp Gly Ala Leu Val Glu
325 330 335Val Met Asn Asn Thr Ser Asp Gly Asp Tyr Gln Thr Phe Lys Ala Asn
340 345 350Asp Gly Ala Tyr Val Arg Glu His Phe Phe Gly Arg Asp Pro Arg Thr
355 360 365Leu Lys Leu Val Glu Asp Met Thr Asp Glu Glu Ile Trp Lys Leu Pro
370 375 380Arg Gly Gly His Asp Tyr Arg Lys Val Tyr Ala Ala Tyr Lys Arg Ala385 390 395 400Leu Glu Thr Lys Asp Arg Pro Thr Val Ile Leu Ala His Thr Ile Lys
405 410 415Gly Tyr Gly Leu Gly His Asn Phe Glu Gly Arg Asn Ala Thr His Gln
420 425 430Met Lys Lys Leu Thr Leu Asp Asp Leu Lys Leu Phe Arg Asp Lys Gln
435 440 445Gly Leu Pro Ile Thr Asp Glu Glu Leu Glu Lys Asp Pro Tyr Leu Pro
450 455 460Pro Tyr Tyr His Pro Gly Glu Asp Ala Pro Glu Ile Lys Tyr Met Lys465 470 475 480Glu Arg Arg Gln Ala Leu Gly Gly Phe Leu Pro Glu Arg Arg Glu Lys
485 490 495Tyr Glu Pro Leu Gln Val Pro Pro Leu Asp Lys Leu Arg Ser Val Arg
500 505 510Lys Gly Ser Gly Lys Gln Gln Val Ala Thr Thr Met Ala Thr Val Arg
515 520 525Thr Phe Lys Glu Leu Met Arg Asp Lys Asn Leu Ala Asp Arg Leu Val
530 535 540Pro Ile Ile Pro Asp Glu Ala Arg Thr Phe Gly Leu Asp Ser Trp Phe545 550 555 560Pro Thr Leu Lys Ile Tyr Asn Pro His Gly Gln Asn Tyr Val Pro Val
565 570 575Asp His Asp Leu Met Leu Ser Tyr Arg Glu Ala Lys Asp Gly Gln Ile
580 585 590Leu His Glu Gly Ile Asn Glu Ala Gly Ser Val Ala Ser Phe Ile Ala
595 600 605Ala Gly Thr Ser Tyr Ala Thr His Gly Glu Ala Met Ile Pro Leu Tyr
610 615 620Ile Phe Tyr Ser Met Phe Gly Phe Gln Arg Thr Gly Asp Gly Ile Trp625 630 635 640Ala Ala Ala Asp Gln Met Thr Arg Gly Phe Leu Leu Gly Ala Thr Ala
645 650 655Gly Arg Thr Thr Leu Thr Gly Glu Gly Leu Gln His Met Asp Gly His
660 665 670Ser Pro Ile Leu Ala Ser Thr Asn Pro Gly Val Glu Thr Tyr Asp Pro
675 680 685Ala Phe Ser Tyr Glu Ile Ala His Leu Val His Arg Gly Ile Asp Arg
690 695 700Met Tyr Gly Pro Gly Lys Gly Glu Asn Val Ile Tyr Tyr Leu Thr Ile705 710 715 720Tyr Asn Glu Pro Thr Pro Gln Pro Ala Glu Pro Glu Asp Leu Asp Val
725 730 735Glu Gly Leu His Lys Gly Ile Tyr Leu Tyr Asp Lys Ala Ala Glu Gly
740 745 750Glu Gly His Glu Ala Ser Ile Leu Ala Ser Gly Ile Gly Met Gln Trp
755 760 765Ala Leu Arg Ala Arg Asp Ile Leu Ala Glu Asp Tyr Gly Ile Arg Ala
770 775 780Asn Ile Phe Ser Ala Thr Ser Trp Val Glu Leu Ala Arg Asp Gly Ala785 790 795 800Arg Arg Asn Leu Glu Ala Leu Arg Asn Pro Gly Ala Asp Val Gly Glu
805 810 815Ala Phe Val Thr Thr Gln Leu Lys Lys Gly Ser Gly Pro Tyr Val Ala
820 825 830Val Ser Asp Phe Ala Thr Asp Leu Pro Asn Gln Ile Arg Glu Trp Val
835 840 845Pro Gly Asp Tyr Ile Val Leu Gly Ala Asp Gly Phe Gly Phe Ser Asp
850 855 860Thr Arg Pro Ala Ala Arg Arg Tyr Phe Asn Ile Asp Ala Glu Ser Ile865 870 875 880Val Val Ala Val Leu Arg Gly Leu Val Arg Glu Gly Val Ile Asp Ala
885 890 895Ser Val Ala Ala His Ala Ala Glu Lys Tyr Lys Leu Ser Asp Pro Thr
900 905 910Ala Pro Gln Val Asp Pro Asp Ala Pro Ile Glu
915 920<210>23<211>4013<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(319)..(3735)<400>23gtcctttttg caaattctgc aaagtgggta gaggtcagat gtcagcaggt cggtccgatt 60tctgtaggaa agtggagccg ttgggggcaa cattaacctt ccccctggga tgtagctaaa 120cggcaatggg ggtctcgggc ggggggcatt cttttcacgg caaggtggtg aaattccgca 180ggtcactccc cggccggcgg tagagaacgg agcgaaaacg gaaagcaata cgtggttttc 240cggactggcc gttacgatgt tctgaagagt gactgccatc acccaacagg ctggtcctcg 300tcgaaaggaa caaaaact gtg gtt aca aca aca ccc tcc acg ctg ccg gcg 351
Val Val Thr Thr Thr Pro Ser Thr Leu Pro Ala
1 5 10ttc aaa aag atc ctg gtg gcc aac cga ggt gaa atc gcg gtg cga gca 399Phe Lys Lys Ile Leu Val Ala Asn Arg Gly Glu Ile Ala Val Arg Ala
15 20 25ttc cgc gcc gcc tac gag acc ggg gcc gca acc gtg gcc atc tac ccc 447Phe Arg Ala Ala Tyr Glu Thr Gly Ala Ala Thr Val Ala Ile Tyr Pro
30 35 40cgg gag gac cgt ggc tcc ttc cac cgc tcc ttc gcc tcc gag gcg gtg 495Arg Glu Asp Arg Gly Ser Phe His Arg Ser Phe Ala Ser Glu Ala Val
45 50 55agg atc gga acc gag ggc tca ccc gtc aag gcg tac ctc gat att gat 543Arg Ile Gly Thr Glu Gly Ser Pro Val Lys Ala Tyr Leu Asp Ile Asp 60 65 70 75gag atc atc aac gcc gcc aag aag gtg aaa gcg gac gcg gtc tac ccg 591Glu Ile Ile Asn Ala Ala Lys Lys Val Lys Ala Asp Ala Val Tyr Pro
80 85 90ggg tat ggt ttc ctt tcg gaa aat gcc cag ctc gcg cgt gaa tgc gcg 639Gly Tyr Gly Phe Leu Ser Glu Asn Ala Gln Leu Ala Arg Glu Cys Ala
95 100 105gag aac ggc att acc ttc atc ggt ccc acc ccg gag gtg ctc gac ctc 687Glu Asn Gly Ile Thr Phe Ile Gly Pro Thr Pro Glu Val Leu Asp Leu
110 115 120acg ggc gac aag tcc aag gct gtg tcc gcc gcg aag aag gcc ggg ctg 735Thr Gly Asp Lys Ser Lys Ala Val Ser Ala Ala Lys Lys Ala Gly Leu
125 130 135ccg gtg ctg gcg gaa tcc acc ccc agc acc gac atc gat gag atc gtc 783Pro Val Leu Ala Glu Ser Thr Pro Ser Thr Asp Ile Asp Glu Ile Val140 145 150 155aag agt gcc gag ggg cag acc tac ccg atc ttc gtc aag gcc gtc gca 831Lys Ser Ala Glu Gly Gln Thr Tyr Pro Ile Phe Val Lys Ala Val Ala
160 165 170ggt ggt ggc ggg cgt ggt atg cgg ttc gtc gag aag ccc gag gac ctg 879Gly Gly Gly Gly Arg Gly Met Arg Phe Val Glu Lys Pro Glu Asp Leu
175 180 185cgt gag ctg gcc agg gag gcc tcc cgc gag gcg gag gcc gct ttc ggt 927Arg Glu Leu Ala Arg Glu Ala Ser Arg Glu Ala Glu Ala Ala Phe Gly
190 195 200gac gga tcc gtc tac gtc gaa cgg gcc gtg atc aaa ccc cag cac atc 975Asp Gly Ser Val Tyr Val Glu Arg Ala Val Ile Lys Pro Gln His Ile
205 210 215gag gtg cag atc ctc ggt gat cac acc ggc gat gtc atc cac ctg tat 1023Glu Val Gln Ile Leu Gly Asp His Thr Gly Asp Val Ile His Leu Tyr220 225 230 235gaa cgc gac tgt tcc ctg cag cgc cgc cac cag aag gtc gtg gag atc 1071Glu Arg Asp Cys Ser Leu Gln Arg Arg His Gln Lys Val Val Glu Ile
240 245 250gca cct gcc cag cac ctc gac ccg gag ctg cgc gac cgc atc tgt gcc 1119Ala Pro Ala Gln His Leu Asp Pro Glu Leu Arg Asp Arg Ile Cys Ala
255 260 265gat gcc gtg aag ttc tgc aaa tcc atc gga tac cag ggc gcc ggc acc 1167Asp Ala Val Lys Phe Cys Lys Ser Ile Gly Tyr Gln Gly Ala Gly Thr
270 275 280gtg gag ttc ctc gtc gac gag gcg ggc aac cac gtc ttc att gag atg 1215Val Glu Phe Leu Val Asp Glu Ala Gly Asn His Val Phe Ile Glu Met
285 290 295aac ccc cgc atc cag gtg gaa cac acc gtg acc gag gag gtc acc tcc 1263Asn Pro Arg Ile Gln Val Glu His Thr Val Thr Glu Glu Val Thr Ser300 305 310 315gtc gac ctg gtc aag gcg cag atg cac ctg gcc gcc ggt gcc acc ctg 1311Val Asp Leu Val Lys Ala Gln Met His Leu Ala Ala Gly Ala Thr Leu
320 325 330aag gaa ctg ggc ctg acc cag gac aag atc acc acc cac ggt gcc gcc 1359Lys Glu Leu Gly Leu Thr Gln Asp Lys Ile Thr Thr His Gly Ala Ala
335 340 345ctg cag tgc cgc atc acc acg gag gac ccg tcc aac aac ttc cgg ccc 1407Leu Gln Cys Arg Ile Thr Thr Glu Asp Pro Ser Asn Asn Phe Arg Pro
350 355 360gac acc ggt gtg atc acc gcc tac cgc tcc ccg ggt ggt gcg ggt gtg 1455Asp Thr Gly Val Ile Thr Ala Tyr Arg Ser Pro Gly Gly Ala Gly Val
365 370 375cgt ctc gac ggc gca gcc cag ctc ggc ggc gag atc acc gca cat ttc 1503Arg Leu Asp Gly Ala Ala Gln Leu Gly Gly Glu Ile Thr Ala His Phe380 385 390 395gat tcc atg ctg gtc aag atg acc tgc cgc ggt tcc gat ttc gag acc 1551Asp Ser Met Leu Val Lys Met Thr Cys Arg Gly Ser Asp Phe Glu Thr
400 405 410gcc gtg tcc cga gcc cag cgc gcc ctg gcg gag ttc aac gtc tcc ggc 1599Ala Val Ser Arg Ala Gln Arg Ala Leu Ala Glu Phe Asn Val Ser Gly
415 420 425gtg gcc acc aac atc ggc ttc ctg cgt gcg ctg ctg cgc gag gaa gac 1647Val Ala Thr Asn Ile Gly Phe Leu Arg Ala Leu Leu Arg Glu Glu Asp
430 435 440ttc acc aag agg cgc atc gac acc ggc ttc atc ggc tcc cac cag cac 1695Phe Thr Lys Arg Arg Ile Asp Thr Gly Phe Ile Gly Ser His Gln His
445 450 455ctg ctc cag gcc cca ccg gcc gac gat gag cag ggg cgg atc ctg gaa 1743Leu Leu Gln Ala Pro Pro Ala Asp Asp Glu Gln Gly Arg Ile Leu Glu460 465 470 475tac ctg gcg gat gtc acc gtg aac aaa ccc cac ggt gaa cgc ccc gag 1791Tyr Leu Ala Asp Val Thr Val Asn Lys Pro His Gly Glu Arg Pro Glu
480 485 490aca gcc cgt ccg ata gag aag ctg ccc gag gtg gag aac atc ccg ctg 1839Thr Ala Arg Pro Ile Glu Lys Leu Pro Glu Val Glu Asn Ile Pro Leu
495 500 505cca cgc ggc tcc cgc gac cgc ctg aag cag ctc ggc ccg gag ggt ttc 1887Pro Arg Gly Ser Arg Asp Arg Leu Lys Gln Leu Gly Pro Glu Gly Phe
510 515 520gcc cgc gat ctg cgc gaa cag gat gcc ctg gcc gtc acc gac acc acc 1935Ala Arg Asp Leu Arg Glu Gln Asp Ala Leu Ala Val Thr Asp Thr Thr
525 530 535ttc cgc gat gcc cac cag tcc ctc ctg gcc acc cgc gtg cgc tcc ttc 1983Phe Arg Asp Ala His Gln Ser Leu Leu Ala Thr Arg Val Arg Ser Phe540 545 550 555gcg ctg acc ccg gcg gcg cgc gcc gtc gca aag ctc acc ccc gag ctg 2031Ala Leu Thr Pro Ala Ala Arg Ala Val Ala Lys Leu Thr Pro Glu Leu
560 565 570ctg tcg gtg gag gcc tgg ggc ggt gcc acc tac gac gtg gcc atg cgc 2079Leu Ser Val Glu Ala Trp Gly Gly Ala Thr Tyr Asp Val Ala Met Arg
575 580 585ttc ctc ttc gag gat ccg tgg gca cgc ctg gat gag ctg cgt gag gcg 2127Phe Leu Phe Glu Asp Pro Trp Ala Arg Leu Asp Glu Leu Arg Glu Ala
590 595 600atg ccg aat gtg aac atc cag atg ctg ctg cgt ggt cgc aac acc gtc 2175Met Pro Asn Val Asn Ile Gln Met Leu Leu Arg Gly Arg Asn Thr Val
605 610 615ggg tac acc ccg tac ccc gat tcg gtg tgc cgc gcg ttt gtg cag gag 2223Gly Tyr Thr Pro Tyr Pro Asp Ser Val Cys Arg Ala Phe Val Gln Glu620 625 630 635gcc gcc aag tcc ggt gtg gac atc ttc cgc atc ttc gac gcg ctc aac 2271Ala Ala Lys Ser Gly Val Asp Ile Phe Arg Ile Phe Asp Ala Leu Asn
640 645 650gac atc tcc cag atg cgc ccg gcc atc gac gcc gtc ctg gag acc ggc 2319Asp Ile Ser Gln Met Arg Pro Ala Ile Asp Ala Val Leu Glu Thr Gly
655 660 665acc agt gtt gcc gag gtc gcc atg gcg tac tcc ggt gac ctg tcc aat 2367Thr Ser Val Ala Glu Val Ala Met Ala Tyr Ser Gly Asp Leu Ser Asn
670 675 680ccg ggg gag aag ctc tac acc ctg gac tac tac ctg aac ctg gcc gag 2415Pro Gly Glu Lys Leu Tyr Thr Leu Asp Tyr Tyr Leu Asn Leu Ala Glu
685 690 695cag atc gtc gac tcc ggt gca cac atc ctg gcc atc aag gac atg gcc 2463Gln Ile Val Asp Ser Gly Ala His Ile Leu Ala Ile Lys Asp Met Ala700 705 710 715ggc ctg ctg cgc cgc gcc gcg gcg ccc aaa ctg gtc acc gcc ctg cgc 2511Gly Leu Leu Arg Arg Ala Ala Ala Pro Lys Leu Val Thr Ala Leu Arg
720 725 730cgt gaa ttc gac ctg ccc gtg cat gtc cac acc cac gac acc gcc ggc 2559Arg Glu Phe Asp Leu Pro Val His Val His Thr His Asp Thr Ala Gly
735 740 745ggt cag ctg gcc acc tac ctg gcc gcc gcc aac gcc ggg gcc gat gcc 2607Gly Gln Leu Ala Thr Tyr Leu Ala Ala Ala Asn Ala Gly Ala Asp Ala
750 755 760gtc gac gcc gcc tcc gca ccc ctg tcc ggt acc acc tcc cag ccg tcg 2655Val Asp Ala Ala Ser Ala Pro Leu Ser Gly Thr Thr Ser Gln Pro Ser
765 770 775atg tcc gct ctg gtt gcc gcg ttt gcg cac acc cga cgc gac acc ggc 2703Met Ser Ala Leu Val Ala Ala Phe Ala His Thr Arg Arg Asp Thr Gly780 785 790 795ctc aac ctg cag gcc gtc tcc gac ctg gaa ccg tac tgg gag gcg gtc 2751Leu Asn Leu Gln Ala Val Ser Asp Leu Glu Pro Tyr Trp Glu Ala Val
800 805 810cgc gga ctg tac ctg ccg ttt gaa tcc ggc acc ccg ggc ccg acc gga 2799Arg Gly Leu Tyr Leu Pro Phe Glu Ser Gly Thr Pro Gly Pro Thr Gly
815 820 825cgc gtt tac cgc cac gag atc ccc ggc ggt cag ctg tcc aac ctg cgt 2847Arg Val Tyr Arg His Glu Ile Pro Gly Gly Gln Leu Ser Asn Leu Arg
830 835 840gcc cag gcc gtt gca ctg ggt ctg gcc gac cgc ttc gag ctc atc gag 2895Ala Gln Ala Val Ala Leu Gly Leu Ala Asp Arg Phe Glu Leu Ile Glu
845 850 855gac tac tac gcg gcc gtc aac gag atg ctg ggt cgt ccg acc aag gtc 2943Asp Tyr Tyr Ala Ala Val Asn Glu Met Leu Gly Arg Pro Thr Lys Val860 865 870 875acc ccg tcc tcc aag gtt gtc ggt gac ctc gca ctg cac ctc gtc ggt 2991Thr Pro Ser Ser Lys Val Val Gly Asp Leu Ala Leu His Leu Val Gly
880 885 890gcc ggt gtg agc ccg gag gat ttc gcc gcc gat ccg cag aag tac gac 3039Ala Gly Val Ser Pro Glu Asp Phe Ala Ala Asp Pro Gln Lys Tyr Asp
895 900 905atc ccc gat tcg gtc atc gcc ttc ctc cgc ggc gaa ctg ggt acc cct 3087Ile Pro Asp Ser Val Ile Ala Phe Leu Arg Gly Glu Leu Gly Thr Pro
910 915 920ccc ggt ggc tgg ccc gaa ccg ctg cgc acc cgt gca ctc gag ggt cgc 3135Pro Gly Gly Trp Pro Glu Pro Leu Arg Thr Arg Ala Leu Glu Gly Arg
925 930 935tcc cag ggt aag gcc ccg ctg gcg gag atc ccc gcc gag gag cag gcc 3183Ser Gln Gly Lys Ala Pro Leu Ala Glu Ile Pro Ala Glu Glu Gln Ala940 945 950 955cac ctg gat tcc gat gat tcc gcg gag cgt cgc ggc acc ctc aac cgc 3231His Leu Asp Ser Asp Asp Ser Ala Glu Arg Arg Gly Thr Leu Asn Arg
960 965 970ctg ctg ttc ccg aag ccg acc gag gag ttc ctt gag cac cgt cgc cgc 3279Leu Leu Phe Pro Lys Pro Thr Glu Glu Phe Leu Glu His Arg Arg Arg
975 980 985ttc ggc aac acc tcc gcc ctg gat gac cgc gag ttc ttc tac ggc ttg 3327Phe Gly Asn Thr Ser Ala Leu Asp Asp Arg Glu Phe Phe Tyr Gly Leu
990 9951000aag gag gga cgt gag gag ctg atc cga ctg acc ggt gtg tcc acc ccg 3375Lys Glu Gly Arg Glu Glu Leu Ile Arg Leu Thr Gly Val Ser Thr Pro 100510101015atg gtg gtc cgc ctg gat gcg gtg tcc gaa ccg gat gac aaa ggc atg 3423Met Val Val Arg Leu Asp Ala Val Ser Glu Pro Asp Asp Lys Gly Met1020 102510301035cgc aac gtg gtg gtc aac gtc aac ggc cag atc cgc ccg atc aag gtg 3471Arg Asn Val Val Val Asn Val Asn Gly Gln Ile Arg Pro Ile Lys Val
104010451050cgc gac cgt tcc gtg gag tcc gtc acc gcc acc gcg gag aag gcc gat 3519Arg Asp Arg Ser Val Glu Ser Val Thr Ala Thr Ala Glu Lys Ala Asp
105510601065gcc acc aac aag ggc cat gtc gcc gca cca ttc gcc ggt gtg gtc acc 3567Ala Thr Asn Lys Gly His Val Ala Ala Pro Phe Ala Gly Val Val Thr
107010751080gtg acc gtc gcc gag ggt gat gag atc aag gct ggc gac gcc gtg gcc 3615Val Thr Val Ala Glu Gly Asp Glu Ile Lys Ala Gly Asp Ala Val Ala 108510901095atc att gag gcc atg aag atg gag gcc acc atc acc gcg cct gtc gac 3663Ile Ile Glu Ala Met Lys Met Glu Ala Thr Ile Thr Ala Pro Val Asp1100 110511101115ggt gtc atc gac cgc gtc gtg gtg ccc gcc gcc acc aag gtc gag ggc 3711Gly Val Ile Asp Arg Val Val Val Pro Ala Ala Thr Lys Val Glu Gly
112011251130ggc gac ctc atc gtg gtc gtg tcc tagcgactga gagccacaac ccgtcccggg 3765Gly Asp Leu Ile Val Val Val Ser
1135tgccttgtta tcaacctccc cctgatgatg ttctcagggg gaggctctac gtacctcacc 3825gtgacggtgc atgtatatcg tcctgctgga gagaatgctc caggtaggaa cgccaaccac 3885cccactccgt gatgtcccgt gctgatccca ggcaggccgg ttggaaagaa aaaccagtga 3945tggaacggcc atcggacagc gagacggaac caagcgtcat cggctccggt agagcggtga 4005ggagcctg 4013<210>24<211>1139<212>PRT<213>Corynebacterium thermoaminogenes<400>24Val Val Thr Thr Thr Pro Ser Thr Leu Pro Ala Phe Lys Lys Ile Leu 1 5 10 15Val Ala Asn Arg Gly Glu Ile Ala Val Arg Ala Phe Arg Ala Ala Tyr
20 25 30Glu Thr Gly Ala Ala Thr Val Ala Ile Tyr Pro Arg Glu Asp Arg Gly
35 40 45Ser Phe His Arg Ser Phe Ala Ser Glu Ala Val Arg Ile Gly Thr Glu
50 55 60Gly Ser Pro Val Lys Ala Tyr Leu Asp Ile Asp Glu Ile Ile Asn Ala 65 70 75 80Ala Lys Lys Val Lys Ala Asp Ala Val Tyr Pro Gly Tyr Gly Phe Leu
85 90 95Ser Glu Asn Ala Gln Leu Ala Arg Glu Cys Ala Glu Asn Gly Ile Thr
100 105 110Phe Ile Gly Pro Thr Pro Glu Val Leu Asp Leu Thr Gly Asp Lys Ser
115 120 125Lys Ala Val Ser Ala Ala Lys Lys Ala Gly Leu Pro Val Leu Ala Glu
130 135 140Ser Thr Pro Ser Thr Asp Ile Asp Glu Ile Val Lys Ser Ala Glu Gly145 150 155 160Gln Thr Tyr Pro Ile Phe Val Lys Ala Val Ala Gly Gly Gly Gly Arg
165 170 175Gly Met Arg Phe Val Glu Lys Pro Glu Asp Leu Arg Glu Leu Ala Arg
180 185 190Glu Ala Ser Arg Glu Ala Glu Ala Ala Phe Gly Asp Gly Ser Val Tyr
195 200 205Val Glu Arg Ala Val Ile Lys Pro Gln His Ile Glu Val Gln Ile Leu
210 215 220Gly Asp His Thr Gly Asp Val Ile His Leu Tyr Glu Arg Asp Cys Ser225 230 235 240Leu Gln Arg Arg His Gln Lys Val Val Glu Ile Ala Pro Ala Gln His
245 250 255Leu Asp Pro Glu Leu Arg Asp Arg Ile Cys Ala Asp Ala Val Lys Phe
260 265 270Cys Lys Ser Ile Gly Tyr Gln Gly Ala Gly Thr Val Glu Phe Leu Val
275 280 285Asp Glu Ala Gly Asn His Val Phe Ile Glu Met Asn Pro Arg Ile Gln
290 295 300Val Glu His Thr Val Thr Glu Glu Val Thr Ser Val Asp Leu Val Lys305 310 315 320Ala Gln Met His Leu Ala Ala Gly Ala Thr Leu Lys Glu Leu Gly Leu
325 330 335Thr Gln Asp Lys Ile Thr Thr His Gly Ala Ala Leu Gln Cys Arg Ile
340 345 350Thr Thr Glu Asp Pro Ser Asn Asn Phe Arg Pro Asp Thr Gly Val Ile
355 360 365Thr Ala Tyr Arg Ser Pro Gly Gly Ala Gly Val Arg Leu Asp Gly Ala
370 375 380Ala Gln Leu Gly Gly Glu Ile Thr Ala His Phe Asp Ser Met Leu Val385 390 395 400Lys Met Thr Cys Arg Gly Ser Asp Phe Glu Thr Ala Val Ser Arg Ala
405 410 415Gln Arg Ala Leu Ala Glu Phe Asn Val Ser Gly Val Ala Thr Asn Ile
420 425 430Gly Phe Leu Arg Ala Leu Leu Arg Glu Glu Asp Phe Thr Lys Arg Arg
435 440 445Ile Asp Thr Gly Phe Ile Gly Ser His Gln His Leu Leu Gln Ala Pro
450 455 460Pro Ala Asp Asp Glu Gln Gly Arg Ile Leu Glu Tyr Leu Ala Asp Val465 470 475 480Thr Val Asn Lys Pro His Gly Glu Arg Pro Glu Thr Ala Arg Pro Ile
485 490 495Glu Lys Leu Pro Glu Val Glu Asn Ile Pro Leu Pro Arg Gly Ser Arg
500 505 510Asp Arg Leu Lys Gln Leu Gly Pro Glu Gly Phe Ala Arg Asp Leu Arg
515 520 525Glu Gln Asp Ala Leu Ala Val Thr Asp Thr Thr Phe Arg Asp Ala His
530 535 540Gln Ser Leu Leu Ala Thr Arg Val Arg Ser Phe Ala Leu Thr Pro Ala545 550 555 560Ala Arg Ala Val Ala Lys Leu Thr Pro Glu Leu Leu Ser Val Glu Ala
565 570 575Trp Gly Gly Ala Thr Tyr Asp Val Ala Met Arg Phe Leu Phe Glu Asp
580 585 590Pro Trp Ala Arg Leu Asp Glu Leu Arg Glu Ala Met Pro Asn Val Asn
595 600 605Ile Gln Met Leu Leu Arg Gly Arg Asn Thr Val Gly Tyr Thr Pro Tyr
610 615 620Pro Asp Ser Val Cys Arg Ala Phe Val Gln Glu Ala Ala Lys Ser Gly625 630 635 640Val Asp Ile Phe Arg Ile Phe Asp Ala Leu Asn Asp Ile Ser Gln Met
645 650 655Arg Pro Ala Ile Asp Ala Val Leu Glu Thr Gly Thr Ser Val Ala Glu
660 665 670Val Ala Met Ala Tyr Ser Gly Asp Leu Ser Asn Pro Gly Glu Lys Leu
675 680 685Tyr Thr Leu Asp Tyr Tyr Leu Asn Leu Ala Glu Gln Ile Val Asp Ser
690 695 700Gly Ala His Ile Leu Ala Ile Lys Asp Met Ala Gly Leu Leu Arg Arg705 710 715 720Ala Ala Ala Pro Lys Leu Val Thr Ala Leu Arg Arg Glu Phe Asp Leu
725 730 735Pro Val His Val His Thr His Asp Thr Ala Gly Gly Gln Leu Ala Thr
740 745 750Tyr Leu Ala Ala Ala Asn Ala Gly Ala Asp Ala Val Asp Ala Ala Ser
755 760 765Ala Pro Leu Ser Gly Thr Thr Ser Gln Pro Ser Met Ser Ala Leu Val
770 775 780Ala Ala Phe Ala His Thr Arg Arg Asp Thr Gly Leu Asn Leu Gln Ala785 790 795 800Val Ser Asp Leu Glu Pro Tyr Trp Glu Ala Val Arg Gly Leu Tyr Leu
805 810 815Pro Phe Glu Ser Gly Thr Pro Gly Pro Thr Gly Arg Val Tyr Arg His
820 825 830Glu Ile Pro Gly Gly Gln Leu Ser Asn Leu Arg Ala Gln Ala Val Ala
835 840 845Leu Gly Leu Ala Asp Arg Phe Glu Leu Ile Glu Asp Tyr Tyr Ala Ala
850 855 860Val Asn Glu Met Leu Gly Arg Pro Thr Lys Val Thr Pro Ser Ser Lys865 870 875 880Val Val Gly Asp Leu Ala Leu His Leu Val Gly Ala Gly Val Ser Pro
885 890 895Glu Asp Phe Ala Ala Asp Pro Gln Lys Tyr Asp Ile Pro Asp Ser Val
900 905 910Ile Ala Phe Leu Arg Gly Glu Leu Gly Thr Pro Pro Gly Gly Trp Pro
915 920 925Glu Pro Leu Arg Thr Arg Ala Leu Glu Gly Arg Ser Gln Gly Lys Ala
930 935 940Pro Leu Ala Glu Ile Pro Ala Glu Glu Gln Ala His Leu Asp Ser Asp945 950 955 960Asp Ser Ala Glu Arg Arg Gly Thr Leu Asn Arg Leu Leu Phe Pro Lys
965 970 975Pro Thr Glu Glu Phe Leu Glu His Arg Arg Arg Phe Gly Asn Thr Ser
980 985 990Ala Leu Asp Asp Arg Glu Phe Phe Tyr Gly Leu Lys Glu Gly Arg Glu
99510001005Glu Leu Ile Arg Leu Thr Gly Val Ser Thr Pro Met Val Val Arg Leu 10101015 1020Asp Ala Val Ser Glu Pro Asp Asp Lys Gly Met Arg Asn Val Val Val1025 103010351040Asn Val Asn Gly Gln Ile Arg Pro Ile Lys Val Arg Asp Arg Ser Val
104510501055Glu Ser Val Thr Ala Thr Ala Glu Lys Ala Asp Ala Thr Asn Lys Gly
106010651070His Val Ala Ala Pro Phe Ala Gly Val Val Thr Val Thr Val Ala Glu
107510801085Gly Asp Glu Ile Lys Ala Gly Asp Ala Val Ala Ile Ile Glu Ala Met 109010951100Lys Met Glu Ala Thr Ile Thr Ala Pro Val Asp Gly Val Ile Asp Arg105111011151120Val Val Val Pro Ala Ala Thr Lys Val Glu Gly Gly Asp Leu Ile Val
112511301135Val Val Ser<210>25<211>3306<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(64)..(2820)<400>25gatcaaccta agccaggaga atccggcggg cggtttctac ttctacagga gctgaacccc 60acc gtg aat gaa ctt ctc cgt gac gat atc cgt tat ctc ggc cgg atc 108
Val Asn Glu Leu Leu Arg Asp Asp Ile Arg Tyr Leu Gly Arg Ile
1 5 10 15ctg ggc gag gtg atc tcc gag cag gag ggc cac cat gtc ttc gaa ctg 156Leu Gly Glu Val Ile Ser Glu Gln Glu Gly His His Val Phe Glu Leu
20 25 30gtt gaa cgc gcc cgc cgg acc tcc ttc gac atc gcc aag gga cgc gcg 204Val Glu Arg Ala Arg Arg Thr Ser Phe Asp Ile Ala Lys Gly Arg Ala
35 40 45gag atg gac agt ctg gtg gag gtg ttc gct ggc atc gac ccg gag gac 252Glu Met Asp Ser Leu Val Glu Val Phe Ala Gly Ile Asp Pro Glu Asp
50 55 60gcc acg ccc gtg gcc cga gcc ttc acc cat ttc gcc ctg ttg gcc aac 300Ala Thr Pro Val Ala Arg Ala Phe Thr His Phe Ala Leu Leu Ala Asn
65 70 75ctc gcg gag gat ttg cat gac gca gcc cag cgg gaa cag gcc ctg aac 348Leu Ala Glu Asp Leu His Asp Ala Ala Gln Arg Glu Gln Ala Leu Asn 80 85 90 95tcg ggt gag ccc gcg ccg gac agc acc ctc gag gcc acc tgg gtg aaa 396Ser Gly Glu Pro Ala Pro Asp Ser Thr Leu Glu Ala Thr Trp Val Lys
100 105 110ctg gat gat gcc ggg gtg ggc agc ggt gag gtc gcc gcg gtg atc cgc 444Leu Asp Asp Ala Gly Val Gly Ser Gly Glu Val Ala Ala Val Ile Arg
115 120 125aat gcg ctc gtc gcc ccg gtg ctc acc gcg cac ccg acg gaa acc cga 492Asn Ala Leu Val Ala Pro Val Leu Thr Ala His Pro Thr Glu Thr Arg
130 135 140cgt cgt acc gtg ttc gac gcg cag aag cac atc acc gcc ctg atg gag 540Arg Arg Thr Val Phe Asp Ala Gln Lys His Ile Thr Ala Leu Met Glu
145 150 155gaa cgc cac ctc ctc ctg gcg ctg ccc acg cat gcc cgg acc cag tcc 588Glu Arg His Leu Leu Leu Ala Leu Pro Thr His Ala Arg Thr Gln Ser160 165 170 175aag ctg gat gac atc gag cgc aac atc cgg cga cgg atc acg atc ctg 636Lys Leu Asp Asp Ile Glu Arg Asn Ile Arg Arg Arg Ile Thr Ile Leu
180 185 190tgg cag acg gcc ctc atc cgt gtg gcc cgt ccc cgc atc gag gat gag 684Trp Gln Thr Ala Leu Ile Arg Val Ala Arg Pro Arg Ile Glu Asp Glu
195 200 205gtc gag gtt gga ctg cgc tac tac aag ctc agc ctg ttg gcc gag atc 732Val Glu Val Gly Leu Arg Tyr Tyr Lys Leu Ser Leu Leu Ala Glu Ile
210 215 220ccc cgc atc aat cat gat gtg acc gtg gaa ctg gcc cgg cgt ttc ggc 780Pro Arg Ile Asn His Asp Val Thr Val Glu Leu Ala Arg Arg Phe Gly
225 230 235ggg gat atc ccc acc acg gcg atg gtc agg ccg gga tcc tgg atc ggc 828Gly Asp Ile Pro Thr Thr Ala Met Val Arg Pro Gly Ser Trp Ile Gly240 245 250 255ggg gac cat gat ggc aac ccc ttc gtc acc gcg gag act gtc acc tac 876Gly Asp His Asp Gly Asn Pro Phe Val Thr Ala Glu Thr Val Thr Tyr
260 265 270gcc acc cat cgg gcc gcg gag acc gtg ctc aag tac tac gtc aag caa 924Ala Thr His Arg Ala Ala Glu Thr Val Leu Lys Tyr Tyr Val Lys Gln
275 280 285ctg cac gcc ctg gaa cac gaa ctc agt ctc tcc gac cgg atg aac gtc 972Leu His Ala Leu Glu His Glu Leu Ser Leu Ser Asp Arg Met Asn Val
290 295 300atc agc gat gag ctg cgt gtg ctt gcc gat gcc ggc cag aat gac atg 1020Ile Ser Asp Glu Leu Arg Val Leu Ala Asp Ala Gly Gln Asn Asp Met
305 310 315ccc agc cgg gtt gat gaa ccc tac cgg cgg gcc atc cac ggc atg cgt 1068Pro Ser Arg Val Asp Glu Pro Tyr Arg Arg Ala Ile His Gly Met Arg320 325 330 335ggc cgg atg ctg gcc acc acg gcc gcc ctg atc ggt gag gag gcg gtc 1116Gly Arg Met Leu Ala Thr Thr Ala Ala Leu Ile Gly Glu Glu Ala Val
340 345 350gag ggc acc tgg ttc aag acc ttc acg ccc tat acc gat acc cac gag 1164Glu Gly Thr Trp Phe Lys Thr Phe Thr Pro Tyr Thr Asp Thr His Glu
355 360 365ttc aaa cgc gac ctc gat atc gtg gat ggt tcc ctg aga atg tcc cgg 1212Phe Lys Arg Asp Leu Asp Ile Val Asp Gly Ser Leu Arg Met Ser Arg
370 375 380gat gac atc atc gcc gat gac cgt ctg gcc atg ctg cgc tcg gcc ctg 1260Asp Asp Ile Ile Ala Asp Asp Arg Leu Ala Met Leu Arg Ser Ala Leu
385 390 395gac agc ttc ggg ttc aac ctc tac tcc ctg gat ctg cgc cag aat tcc 1308Asp Ser Phe Gly Phe Asn Leu Tyr Ser Leu Asp Leu Arg Gln Asn Ser400 405 410 415gac ggt ttc gag gat gtc ctc acc gaa ttg ttc gcc acc gcc cag acc 1356Asp Gly Phe Glu Asp Val Leu Thr Glu Leu Phe Ala Thr Ala Gln Thr
420 425 430gag aag aac tac cgc ggg ttg acg gag gcg gag aag ctg gac ctg ctg 1404Glu Lys Asn Tyr Arg Gly Leu Thr Glu Ala Glu Lys Leu Asp Leu Leu
435 440 445atc cgc gaa ctg agc aca ccc cgc ccg ctc atc ccg cac ggg gac ccg 1452Ile Arg Glu Leu Ser Thr Pro Arg Pro Leu Ile Pro His Gly Asp Pro
450 455 460gac tac tcc gag gcc acc aac cgt gaa ctg ggg att ttt tcg aag gcc 1500Asp Tyr Ser Glu Ala Thr Asn Arg Glu Leu Gly Ile Phe Ser Lys Ala
465 470 475gcg gag gcc gtg cgt aaa ttc ggt cct ctc atg gtg ccg cac tgc atc 1548Ala Glu Ala Val Arg Lys Phe Gly Pro Leu Met Val Pro His Cys Ile480 485 490 495atc tcc atg gcc tct tcc gtc acg gac ate ctc gaa ccg atg gtg ctg 1596Ile Ser Met Ala Ser Ser Val Thr Asp Ile Leu Glu Pro Met Val Leu
500 505 510ctc aag gag ttc ggt ctg atc cgg gcc aac ggg aag aac ccg acg ggc 1644Leu Lys Glu Phe Gly Leu Ile Arg Ala Asn Gly Lys Asn Pro Thr Gly
515 520 525agc gtc gac gtg atc ccg ctg ttc gag acg atc gat gac ctc cag cgt 1692Ser Val Asp Val Ile Pro Leu Phe Glu Thr Ile Asp Asp Leu Gln Arg
530 535 540ggc gcg ggc atc ctg gag gaa ttg tgg gac atc gac ctc tac cgc aat 1740Gly Ala Gly Ile Leu Glu Glu Leu Trp Asp Ile Asp Leu Tyr Arg Asn
545 550 555tac ctt gag cag cgg gac aac gtc cag gag gtc atg ctg ggg tat tcc 1788Tyr Leu Glu Gln Arg Asp Asn Val Gln Glu Val Met Leu Gly Tyr Ser560 565 570 575gac tcc aac aag gac ggc ggg tac ttc gcc gcc aac tgg gcg ctt tac 1836Asp Ser Asn Lys Asp Gly Gly Tyr Phe Ala Ala Asn Trp Ala Leu Tyr
580 585 590gac gcg gag tta cgc ctg gtc gaa cta tgc cgg ggc cgt aat gtc aag 1884Asp Ala Glu Leu Arg Leu Val Glu Leu Cys Arg Gly Arg Asn Val Lys
595 600 605ctc cgt ctc ttc cac ggt cgt ggt ggc acg gtg ggt cgt ggc ggt ggc 1932Leu Arg Leu Phe His Gly Arg Gly Gly Thr Val Gly Arg Gly Gly Gly
610 615 620ccc tcc tat gat gcg atc ctg gcc cag ccc aag ggc gcg gtc cgg ggt 1980Pro Ser Tyr Asp Ala Ile Leu Ala Gln Pro Lys Gly Ala Val Arg Gly
625 630 635gcg gtg cgg gtg act gaa cag ggc gag atc atc tcc gcg aag tac ggt 2028Ala Val Arg Val Thr Glu Gln Gly Glu Ile Ile Ser Ala Lys Tyr Gly640 645 650 655aac ccg gat acg gca cgc cgc aac ctt gag gcc ctg gtg tcc gcg acg 2076Asn Pro Asp Thr Ala Arg Arg Asn Leu Glu Ala Leu Val Ser Ala Thr
660 665 670ctg gag gca tcg ctt ctg gat gat gtg gaa ctg ccc aat cgg gaa cgc 2124Leu Glu Ala Ser Leu Leu Asp Asp Val Glu Leu Pro Asn Arg Glu Arg
675 680 685gcg cac cag atc atg ggg gag atc tcg gag ttg agc ttc cgc agg tac 2172Ala His Gln Ile Met Gly Glu Ile Ser Glu Leu Ser Phe Arg Arg Tyr
690 695 700tca tca ctg gtc cat gag gat ccc gga ttc atc cag tac ttc acc cag 2220Ser Ser Leu Val His Glu Asp Pro Gly Phe Ile Gln Tyr Phe Thr Gln
705 710 715tcc acc ccc ctg cag gag atc gga tcc ctc aac atc ggt tcc cga ccc 2268Ser Thr Pro Leu Gln Glu Ile Gly Ser Leu Asn Ile Gly Ser Arg Pro720 725 730 735tcc tca cgt aaa cag acc aac acg gtg gag gat ctg cgt gcc atc ccg 2316Ser Ser Arg Lys Gln Thr Asn Thr Val Glu Asp Leu Arg Ala Ile Pro
740 745 750tgg gtg ctc agc tgg tcc cag tcc cgt gtc atg ctg ccg ggc tgg ttc 2364Trp Val Leu Ser Trp Ser Gln Ser Arg Val Met Leu Pro Gly Trp Phe
755 760 765ggt gtg ggt acc gca ctg cgt gag tgg atc ggt gag ggg gag ggg gct 2412Gly Val Gly Thr Ala Leu Arg Glu Trp Ile Gly Glu Gly Glu Gly Ala
770 775 780gcg gag cgc atc gcg gag ctg cag gaa ctc aac cgg tgc tgg ccg ttc 2460Ala Glu Arg Ile Ala Glu Leu Gln Glu Leu Asn Arg Cys Trp Pro Phe
785 790 795ttc acc tcg gtg ctg gac aac atg gcc cag gtg atg agc aag gcg gaa 2508Phe Thr Ser Val Leu Asp Asn Met Ala Gln Val Met Ser Lys Ala Glu800 805 810 815ctg cgc ctg gcc agg ttg tac gcc gat ctc atc ccg gat cgc gag gtg 2556Leu Arg Leu Ala Arg Leu Tyr Ala Asp Leu Ile Pro Asp Arg Glu Val
820 825 830gcg gac cgg atc tat gag acc atc ttc ggg gag tat ttc ctg acc aag 2604Ala Asp Arg Ile Tyr Glu Thr Ile Phe Gly Glu Tyr Phe Leu Thr Lys
835 840 845gag atg ttc tgc acc atc acc ggt tcc cag gac ctg ctc gat gac aac 2652Glu Met Phe Cys Thr Ile Thr Gly Ser Gln Asp Leu Leu Asp Asp Asn
850 855 860ccg gcg ctg gcg cga tcg gtg cgc agt cgg ttc ccg tac ctg ctg ccg 2700Pro Ala Leu Ala Arg Ser Val Arg Ser Arg Phe Pro Tyr Leu Leu Pro
865 870 875ctc aat gtc atc cag gtg gag atg atg cgc cgg tac cgg tcc ggt gat 2748Leu Asn Val Ile Gln Val Glu Met Met Arg Arg Tyr Arg Ser Gly Asp880 885 890 895gag ggc acg gct gtc cca cgt aat atc cgc ctg acc atg aat gga ttg 2796Glu Gly Thr Ala Val Pro Arg Asn Ile Arg Leu Thr Met Asn Gly Leu
900 905 910tcc acg gcc ctg cgc aac tcg ggt tagggcgcca gacgccccgg gaacccgcac 2850Ser Thr Ala Leu Arg Asn Ser Gly
915cctgtgtata ctgtctaaag ttgcccggtg tcatccgggc gtgatggata gacaacttaa 2910cggcaaagga ttctccccac atggcactga cgcttcaaat cgtcctcgtt ctcgccagcg 2970tgctcatgac ggtcttcgtc ctgctgcaca agggtaaggg cggaggtctg tcaagcctct 3030tcggtggtgg cgtccagtcc aacctctccg gttccacggt ggtggagaag aacctggacc 3090gcgtcaccat cctgaccgca gtcatctggt tgatctgcat tgtcgcgctc aacctcatcc 3150aggcgtactc ctagcacctg atctttcaag gcctgccctt cggggcaggc cttttttgca 3210ttctccaggt gatgtccatc acccaccggt tttaaactat tgaccgatag aaacacctgc 3270actaggttat ctgttatgca atagaaaata gtgcat 3306<210>26<211>919<212>PRT<213>Corynebacterium thermoaminogenes<400>26Val Asn Glu Leu Leu Arg Asp Asp Ile Arg Tyr Leu Gly Arg Ile Leu 1 5 10 15Gly Glu Val Ile Ser Glu Gln Glu Gly His His Val Phe Glu Leu Val
20 25 30Glu Arg Ala Arg Arg Thr Ser Phe Asp Ile Ala Lys Gly Arg Ala Glu
35 40 45Met Asp Ser Leu Val Glu Val Phe Ala Gly Ile Asp Pro Glu Asp Ala
50 55 60Thr Pro Val Ala Arg Ala Phe Thr His Phe Ala Leu Leu Ala Asn Leu 65 70 75 80Ala Glu Asp Leu His Asp Ala Ala Gln Arg Glu Gln Ala Leu Asn Ser
85 90 95Gly Glu Pro Ala Pro Asp Ser Thr Leu Glu Ala Thr Trp Val Lys Leu
100 105 110Asp Asp Ala Gly Val Gly Ser Gly Glu Val Ala Ala Val Ile Arg Asn
115 120 125Ala Leu Val Ala Pro Val Leu Thr Ala His Pro Thr Glu Thr Arg Arg
130 135 140Arg Thr Val Phe Asp Ala Gln Lys His Ile Thr Ala Leu Met Glu Glu145 150 155 160Arg His Leu Leu Leu Ala Leu Pro Thr His Ala Arg Thr Gln Ser Lys
165 170 175Leu Asp Asp Ile Glu Arg Asn Ile Arg Arg Arg Ile Thr Ile Leu Trp
180 185 190Gln Thr Ala Leu Ile Arg Val Ala Arg Pro Arg Ile Glu Asp Glu Val
195 200 205Glu Val Gly Leu Arg Tyr Tyr Lys Leu Ser Leu Leu Ala Glu Ile Pro
210 215 220Arg Ile Asn His Asp Val Thr Val Glu Leu Ala Arg Arg Phe Gly Gly225 230 235 240Asp Ile Pro Thr Thr Ala Met Val Arg Pro Gly Ser Trp Ile Gly Gly
245 250 255Asp His Asp Gly Asn Pro Phe Val Thr Ala Glu Thr Val Thr Tyr Ala
260 265 270Thr His Arg Ala Ala Glu Thr Val Leu Lys Tyr Tyr Val Lys Gln Leu
275 280 285His Ala Leu Glu His Glu Leu Ser Leu Ser Asp Arg Met Asn Val Ile
290 295 300Ser Asp Glu Leu Arg Val Leu Ala Asp Ala Gly Gln Asn Asp Met Pro305 310 315 320Ser Arg Val Asp Glu Pro Tyr Arg Arg Ala Ile His Gly Met Arg Gly
325 330 335Arg Met Leu Ala Thr Thr Ala Ala Leu Ile Gly Glu Glu Ala Val Glu
340 345 350Gly Thr Trp Phe Lys Thr Phe Thr Pro Tyr Thr Asp Thr His Glu Phe
355 360 365Lys Arg Asp Leu Asp Ile Val Asp Gly Ser Leu Arg Met Ser Arg Asp
370 375 380Asp Ile Ile Ala Asp Asp Arg Leu Ala Met Leu Arg Ser Ala Leu Asp385 390 395 400Ser Phe Gly Phe Asn Leu Tyr Ser Leu Asp Leu Arg Gln Asn Ser Asp
405 410 415Gly Phe Glu Asp Val Leu Thr Glu Leu Phe Ala Thr Ala Gln Thr Glu
420 425 430Lys Asn Tyr Arg Gly Leu Thr Glu Ala Glu Lys Leu Asp Leu Leu Ile
435 440 445Arg Glu Leu Ser Thr Pro Arg Pro Leu Ile Pro His Gly Asp Pro Asp
450 455 460Tyr Ser Glu Ala Thr Asn Arg Glu Leu Gly Ile Phe Ser Lys Ala Ala465 470 475 480Glu Ala Val Arg Lys Phe Gly Pro Leu Met Val Pro His Cys Ile Ile
485 490 495Ser Met Ala Ser Ser Val Thr Asp Ile Leu Glu Pro Met Val Leu Leu
500 505 510Lys Glu Phe Gly Leu Ile Arg Ala Asn Gly Lys Asn Pro Thr Gly Ser
515 520 525Val Asp Val Ile Pro Leu Phe Glu Thr Ile Asp Asp Leu Gln Arg Gly
530 535 540Ala Gly Ile Leu Glu Glu Leu Trp Asp Ile Asp Leu Tyr Arg Asn Tyr545 550 555 560Leu Glu Gln Arg Asp Asn Val Gln Glu Val Met Leu Gly Tyr Ser Asp
565 570 575Ser Asn Lys Asp Gly Gly Tyr Phe Ala Ala Asn Trp Ala Leu Tyr Asp
580 585 590Ala Glu Leu Arg Leu Val Glu Leu Cys Arg Gly Arg Asn Val Lys Leu
595 600 605Arg Leu Phe His Gly Arg Gly Gly Thr Val Gly Arg Gly Gly Gly Pro
610 615 620Ser Tyr Asp Ala Ile Leu Ala Gln Pro Lys Gly Ala Val Arg Gly Ala625 630 635 640Val Arg Val Thr Glu Gln Gly Glu Ile Ile Ser Ala Lys Tyr Gly Asn
645 650 655Pro Asp Thr Ala Arg Arg Asn Leu Glu Ala Leu Val Ser Ala Thr Leu
660 665 670Glu Ala Ser Leu Leu Asp Asp Val Glu Leu Pro Asn Arg Glu Arg Ala
675 680 685His Gln Ile Met Gly Glu Ile Ser Glu Leu Ser Phe Arg Arg Tyr Ser
690 695 700Ser Leu Val His Glu Asp Pro Gly Phe Ile Gln Tyr Phe Thr Gln Ser705 710 715 720Thr Pro Leu Gln Glu Ile Gly Ser Leu Asn Ile Gly Ser Arg Pro Ser
725 730 735Ser Arg Lys Gln Thr Asn Thr Val Glu Asp Leu Arg Ala Ile Pro Trp
740 745 750Val Leu Ser Trp Ser Gln Ser Arg Val Met Leu Pro Gly Trp Phe Gly
755 760 765Val Gly Thr Ala Leu Arg Glu Trp Ile Gly Glu Gly Glu Gly Ala Ala
770 775 780Glu Arg Ile Ala Glu Leu Gln Glu Leu Asn Arg Cys Trp Pro Phe Phe785 790 795 800Thr Ser Val Leu Asp Asn Met Ala Gln Val Met Ser Lys Ala Glu Leu
805 810 815Arg Leu Ala Arg Leu Tyr Ala Asp Leu Ile Pro Asp Arg Glu Val Ala
820 825 830Asp Arg Ile Tyr Glu Thr Ile Phe Gly Glu Tyr Phe Leu Thr Lys Glu
835 840 845Met Phe Cys Thr Ile Thr Gly Ser Gln Asp Leu Leu Asp Asp Asn Pro
850 855 860Ala Leu Ala Arg Ser Val Arg Ser Arg Phe Pro Tyr Leu Leu Pro Leu865 870 875 880Asn Val Ile Gln Val Glu Met Met Arg Arg Tyr Arg Ser Gly Asp Glu
885 890 895Gly Thr Ala Val Pro Arg Asn Ile Arg Leu Thr Met Asn Gly Leu Ser
900 905 910Thr Ala Leu Arg Asn Ser Gly
915<210>27<211>3907<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(686)..(3388)<400>27attacttcag ctgactcagc aacattcgta ttaggtatgc aaacaacatt tggttcgtta 60aatccaagta gtatggttaa agtaacttgg ggtattgctc aagcacttat cgcctttgta 120ttattattag ctggtggcgg agatggaact aaagctctca acgcaattca gagtgccgct 180attattagtg cgtttccatt ctcctttgtc gtcatattaa tgatgatcag tttctacaaa 240gatgctaata aagaacgtaa attcttagga ttaacattaa cgcctaataa acacagatta 300gaagaatacg ttaaatatca acaagaggat tacgaatctg atattttaga aaaacgtgaa 360tctagacgta atcgtgaaag agaagaataa ttgaatgaaa tatctactat aatggtgggt 420ttaaagctat caacaatttt gttgatagct atttttatgt ttcaaacata taaatattat 480ttacttgcga ttgataacca ttctcaatta ataaaaataa cttatagtac aaatgcgtta 540taataagttt tacttatact acctgattaa aaatgcgaaa tgaaaaatga cccctttata 600tacctataca gttgtgttcg aaaacatata ataatacaat ttaactaagg catataaata 660tatagaaatt caagggggat atcaa atg gct tct aat ttt aaa gaa aca gcg 712
Met Ala Ser Asn Phe Lys Glu Thr Ala
1 5aag aaa caa ttt gat tta aat ggc caa tca tac acg tac tat gat tta 760Lys Lys Gln Phe Asp Leu Asn Gly Gln Ser Tyr Thr Tyr Tyr Asp Leu 10 15 20 25aaa tca tta gaa gaa caa ggt tta act aaa att tca aag tta cct tat 808Lys Ser Leu Glu Glu Gln Gly Leu Thr Lys Ile Ser Lys Leu Pro Tyr
30 35 40tca atc cgt gta tta cta gaa tca gtg tta cgt cag gaa gat gat ttt 856Ser Ile Arg Val Leu Leu Glu Ser Val Leu Arg Gln Glu Asp Asp Phe
45 50 55gta att act gat gat cac att aaa caa tta gca gaa ttt ggc aaa aaa 904Val Ile Thr Asp Asp His Ile Lys Gln Leu Ala Glu Phe Gly Lys Lys
60 65 70ggt aac gaa ggt gaa gta cct ttc aaa cca tct cga gtt att tta caa 952Gly Asn Glu Gly Glu Val Pro Phe Lys Pro Scr Arg Val Ile Leu Gln
75 80 85gac ttc act ggt gta cca gca gta gtt gac tta gcg tct tta cgt aaa 1000Asp Phe Thr Gly Val Pro Ala Val Val Asp Leu Ala Ser Leu Arg Lys 90 95 100 105gca atg aat gat gtt ggt ggg gat att aat aaa att aac cct gaa gta 1048Ala Met Asn Asp Val Gly Gly Asp Ile Asn Lys Ile Asn Pro Glu Val
110 115 120cca gtt gac tta gtt att gac cac tct gta caa gta gat agt tat gct 1096Pro Val Asp Leu Val Ile Asp His Ser Val Gln Val Asp Ser Tyr Ala
125 130 135aat cca gat gca tta caa cgt aac atg aaa tta gaa ttt gaa cgt aac 1144Asn Pro Asp Ala Leu Gln Arg Asn Met Lys Leu Glu Phe Glu Arg Asn
140 145 150tat gaa cgt tac caa ttc tta aac tgg gca aca aaa gca ttt gat aac 1192Tyr Glu Arg Tyr Gln Phe Leu Asn Trp Ala Thr Lys Ala Phe Asp Asn
155 160 165tat aat gca gta cca cct gct aca ggt att gtc cac caa gta aac tta 1240Tyr Asn Ala Val Pro Pro Ala Thr Gly Ile Val His Gln Val Asn Leu170 175 180 185gaa tac tta gcg aat gtt gta cat gtt cgt gac gtt gac gga gaa caa 1288Glu Tyr Leu Ala Asn Val Val His Val Arg Asp Val Asp Gly Glu Gln
190 195 200act gct ttc cca gat aca tta gtt ggt act gac tca cat act aca atg 1336Thr Ala Phe Pro Asp Thr Leu Val Gly Thr Asp Ser His Thr Thr Met
205 210 215att aac ggt att ggt gta tta ggt tgg ggt gtc ggc ggt atc gaa gct 1384Ile Asn Gly Ile Gly Val Leu Gly Trp Gly Val Gly Gly Ile Glu Ala
220 225 230gaa gca ggt atg tta gga caa cca tca tac ttc cca att cca gaa gtt 1432Glu Ala Gly Met Leu Gly Gln Pro Ser Tyr Phe Pro Ile Pro Glu Val
235 240 245att ggt gtt aaa tta agt aat gaa tta cca caa ggt tca aca gca act 1480Ile Gly Val Lys Leu Ser Asn Glu Leu Pro Gln Gly Ser Thr Ala Thr250 255 260 265gac tta gca tta cgt gta act gaa gag tta cgt aaa cgt ggt gta gta 1528Asp Leu Ala Leu Arg Val Thr Glu Glu Leu Arg Lys Arg Gly Val Val
270 275 280ggt aaa ttc gtt gag ttc ttt ggt cct ggt gta aca aac tta cca tta 1576Gly Lys Phe Val Glu Phe Phe Gly Pro Gly Val Thr Asn Leu Pro Leu
285 290 295gct gac cgt gca aca att gcg aac atg gcg cct gaa tat ggt gca act 1624Ala Asp Arg Ala Thr Ile Ala Asn Met Ala Pro Glu Tyr Gly Ala Thr
300 305 310tgt ggt ttc ttc cca gtt gat gaa gaa tca ctt aaa tac atg aaa tta 1672Cys Gly Phe Phe Pro Val Asp Glu Glu Ser Leu Lys Tyr Met Lys Leu
315 320 325act ggt cgt aaa gat gat cat att gca cta gta aaa gaa tat tta caa 1720Thr Gly Arg Lys Asp Asp His Ile Ala Leu Val Lys Glu Tyr Leu Gln330 335 340 345caa aat aat atg ttc ttc caa gtt gaa aat gaa gat cct gaa tat act 1768Gln Asn Asn Met Phe Phe Gln Val Glu Asn Glu Asp Pro Glu Tyr Thr
350 355 360gaa gtg att gat tta gat tta tct aca gtt caa gct tct tta tca ggt 1816Glu Val Ile Asp Leu Asp Leu Ser Thr Val Gln Ala Ser Leu Ser Gly
365 370 375cca aaa cgt cca caa gat tta atc ttc tta agt gac atg aaa act gaa 1864Pro Lys Arg Pro Gln Asp Leu Ile Phe Leu Ser Asp Met Lys Thr Glu
380 385 390ttc gaa aaa tca gtt aca gca cca gct ggt aac caa ggt cac ggt tta 1912Phe Glu Lys Ser Val Thr Ala Pro Ala Gly Asn Gln Gly His Gly Leu
395 400 405gat gaa agt gaa ttt gat aag aaa gca gaa atc aaa ttt aat gat ggt 1960Asp Glu Ser Glu Phe Asp Lys Lys Ala Glu Ile Lys Phe Asn Asp Gly410 415 420 425aga act tca act atg aag act ggt gat gtt gcg att gca gcg att aca 2008Arg Thr Ser Thr Met Lys Thr Gly Asp Val Ala Ile Ala Ala Ile Thr
430 435 440tca tgt aca aat aca tct aac cct tac gtt atg tta ggt gca ggt tta 2056Ser Cys Thr Asn Thr Ser Asn Pro Tyr Val Met Leu Gly Ala Gly Leu
445 450 455gta get aaa aaa gca att gaa aaa ggc tta aaa gta cct gat tat gta 2104Val Ala Lys Lys Ala Ile Glu Lys Gly Leu Lys Val Pro Asp Tyr Val
460 465 470aaa act tca tta gca cca ggt tca aaa gtt gtt act gga tat tta aga 2152Lys Thr Ser Leu Ala Pro Gly Ser Lys Val Val Thr Gly Tyr Leu Arg
475 480 485gat tca ggt tta caa gaa tat ctt gat gac tta ggt ttc aac tta gtt 2200Asp Ser Gly Leu Gln Glu Tyr Leu Asp Asp Leu Gly Phe Asn Leu Val490 495 500 505ggt tat ggt tgt aca act tgt atc ggt aac tca ggt cca tta tta cct 2248Gly Tyr Gly Cys Thr Thr Cys Ile Gly Asn Ser Gly Pro Leu Leu Pro
510 515 520gaa att gaa aaa gca gta gct gac gaa gat tta tta gta act tct gta 2296Glu Ile Glu Lys Ala Val Ala Asp Glu Asp Leu Leu Val Thr Ser Val
525 530 535ctt tct ggt aac cgt aac ttt gaa ggt cgt atc cat ccg tta gtt aaa 2344Leu Ser Gly Asn Arg Asn Phe Glu Gly Arg Ile His Pro Leu Val Lys
540 545 550gct aac tac tta gct tca cca caa tta gtt gta gct tat gca tta gct 2392Ala Asn Tyr Leu Ala Ser Pro Gin Leu Val Val Ala Tyr Ala Leu Ala
555 560 565gga acg gtt gat atc gat tta cac aat gaa cct atc ggt aaa ggt aaa 2440Gly Thr Val Asp Ile Asp Leu His Asn Glu Pro Ile Gly Lys Gly Lys570 575 580 585gat ggc gaa gat gta tac ctt aaa gat atc tgg cca agt atc aaa gaa 2488Asp Gly Glu Asp Val Tyr Leu Lys Asp Ile Trp Pro Ser Ile Lys Glu
590 595 600gtt gca gac act gtt gat agt gtc gta acg cca gaa tta ttc tta gaa 2536Val Ala Asp Thr Val Asp Ser Val Val Thr Pro Glu Leu Phe Leu Glu
605 610 615gaa tat gca aat gta tac gaa aat aat gaa atg tgg aat gaa atc gac 2584Glu Tyr Ala Asn Val Tyr Glu Asn Asn Glu Met Trp Asn Glu Ile Asp
620 625 630gtt act gac gca cca tta tat gat ttc gat cca aat tca act tat att 2632Val Thr Asp Ala Pro Leu Tyr Asp Phe Asp Pro Asn Ser Thr Tyr Ile
635 640 645caa aat cca tca ttc ttc caa ggt tta tct aaa gaa cca gga act att 2680Gln Asn Pro Ser Phe Phe Gln Gly Leu Ser Lys Glu Pro Gly Thr Ile650 655 660 665gaa cca tta aaa gat tta cgt att atg ggt aaa ttt ggt gat tca gtt 2728Glu Pro Leu Lys Asp Leu Arg Ile Met Gly Lys Phe Gly Asp Ser Val
670 675 680aca act gac cac att tct cca gca ggt gcg atc ggt aaa gat aca cca 2776Thr Thr Asp His Ile Ser Pro Ala Gly Ala Ile Gly Lys Asp Thr Pro
685 690 695gca ggt aaa tat tta tta gac cat gat gtt cca att aga gaa ttt aac 2824Ala Gly Lys Tyr Leu Leu Asp His Asp Val Pro Ile Arg Glu Phe Asn
700 705 710tct tat ggt tca aga cgt ggt aac cat gaa gta atg gta cgt ggt act 2872Ser Tyr Gly Ser Arg Arg Gly Asn His Glu Val Met Val Arg Gly Thr
715 720 725ttc gct aat atc cgt att aaa aac caa tta gca cca ggc act gaa ggt 2920Phe Ala Asn Ile Arg Ile Lys Asn Gln Leu Ala Pro Gly Thr Glu Gly730 735 740 745gga ttt aca aca tat tgg cct aca gaa gaa atc atg cct atc tat gat 2968Gly Phe Thr Thr Tyr Trp Pro Thr Glu Glu Ile Met Pro Ile Tyr Asp
750 755 760gca gct atg aga tac aaa gaa aat ggt act ggt tta gct gtt tta gct 3016Ala Ala Met Arg Tyr Lys Glu Asn Gly Thr Gly Leu Ala Val Leu Ala
765 770 775ggt aat gat tac ggt atg ggt tca tct cgt gac tgg gca gct aaa ggt 3064Gly Asn Asp Tyr Gly Met Gly Ser Ser Arg Asp Trp Ala Ala Lys Gly
780 785 790act aac tta tta ggt gtt aaa act gtt att gca caa agt tat gaa cgt 3112Thr Asn Leu Leu Gly Val Lys Thr Val Ile Ala Gln Ser Tyr Glu Arg
795 800 805atc cat cgt tca aac tta gta atg atg ggt gta tta cca tta caa ttt 3160Ile His Arg Ser Asn Leu Val Met Met Gly Val Leu Pro Leu Gln Phe810 815 820 825aaa caa ggt gag tca gct gat tct cta ggt tta gaa ggt aaa gaa gaa 3208Lys Gln Gly Glu Ser Ala Asp Ser Leu Gly Leu Glu Gly Lys Glu Glu
830 835 840att tct gta gat atc gat gaa aat gtt aaa cca cat gat tta gta act 3256Ile Ser Val Asp Ile Asp Glu Asn Val Lys Pro His Asp Leu Val Thr
845 850 855gtt cat gct aaa aaa gaa aac gga gaa gtt gtt gat ttt gaa gca atg 3304Val His Ala Lys Lys Glu Asn Gly Glu Val Val Asp Phe Glu Ala Met
860 865 870gtt cgt ttc gat tca tta gta gaa tta gat tat tat cgt cat ggt ggt 3352Val Arg Phe Asp Ser Leu Val Glu Leu Asp Tyr Tyr Arg His Gly Gly
875 880 885atc tta caa atg gta tta aga aac aaa tta gct caa taatcacaat3398Ile Leu Gln Met Val Leu Arg Asn Lys Leu Ala Gln890 895 900gtgacttttg acagtgctaa cgtttaggtt agcactgttt ttttatgcta aactatatat 3458gtaatgttaa tagttaagga aggattggac ttaaatgatt tatagtttga ctgaaattga 3518accaagatat caagagacag ataaaatggg cgtgatttat catggcaatt atgcaacatg 3578gtttgaagta gcgcgtacag attacattag aaaactagga tttagttatg ctgatatgga 3638aaagcaaggg atcatttctc cagttacaga cttaaatatc aaatataaaa aatcaatttt 3698ttatcctgaa aaagtaacca ttaaaacatg ggtggaaaaa tattcaagat tacgttctgt 3758gtatagatat gaaattttta atgaacaggg agaacttgca actacaggtt atactgagtt 3818aatttgtatg aaagctgata cctttagacc aattagatta gatcgttatt tctcagattg 3878gcatgaaacc tatagtaaag ttgaagctt 3907<210>28<211>901<212>PRT<213>Corynebacterium thermoaminogenes<400>28Met Ala Ser Asn Phe Lys Glu Thr Ala Lys Lys Gln Phe Asp Leu Asn 1 5 10 15Gly Gln Ser Tyr Thr Tyr Tyr Asp Leu Lys Ser Leu Glu Glu Gln Gly
20 25 30Leu Thr Lys Ile Ser Lys Leu Pro Tyr Ser Ile Arg Val Leu Leu Glu
35 40 45Ser Val Leu Arg Gln Glu Asp Asp Phe Val Ile Thr Asp Asp His Ile
50 55 60Lys Gln Leu Ala Glu Phe Gly Lys Lys Gly Asn Glu Gly Glu Val Pro 65 70 75 80Phe Lys Pro Ser Arg Val Ile Leu Gln Asp Phe Thr Gly Val Pro Ala
85 90 95Val Val Asp Leu Ala Ser Leu Arg Lys Ala Met Asn Asp Val Gly Gly
100 105 110Asp Ile Asn Lys Ile Asn Pro Glu Val Pro Val Asp Leu Val Ile Asp
115 120 125His Ser Val Gln Val Asp Ser Tyr Ala Asn Pro Asp Ala Leu Gln Arg
130135 140Asn Met Lys Leu Glu Phe Glu Arg Asn Tyr Glu Arg Tyr Gln Phe Leu145 150 155 160Asn Trp Ala Thr Lys Ala Phe Asp Asn Tyr Asn Ala Val Pro Pro Ala
165 170 175Thr Gly Ile Val His Gln Val Asn Leu Glu Tyr Leu Ala Asn Val Val
180 185 190His Val Arg Asp Val Asp Gly Glu Gln Thr Ala Phe Pro Asp Thr Leu
195 200 205Val Gly Thr Asp Ser His Thr Thr Met Ile Asn Gly Ile Gly Val Leu
210 215 220Gly Trp Gly Val Gly Gly Ile Glu Ala Glu Ala Gly Met Leu Gly Gln225 230 235 240Pro Ser Tyr Phe Pro Ile Pro Glu Val Ile Gly Val Lys Leu Ser Asn
245 250 255Glu Leu Pro Gln Gly Ser Thr Ala Thr Asp Leu Ala Leu Arg Val Thr
260 265 270Glu Glu Leu Arg Lys Arg Gly Val Val Gly Lys Phe Val Glu Phe Phe
275 280 285Gly Pro Gly Val Thr Asn Leu Pro Leu Ala Asp Arg Ala Thr Ile Ala
290 295 300Asn Met Ala Pro Glu Tyr Gly Ala Thr Cys Gly Phe Phe Pro Val Asp305 310 315 320Glu Glu Ser Leu Lys Tyr Met Lys Leu Thr Gly Arg Lys Asp Asp His
325 330 335Ile Ala Leu Val Lys Glu Tyr Leu Gln Gln Asn Asn Met Phe Phe Gln
340 345 350Val Glu Asn Glu Asp Pro Glu Tyr Thr Glu Val Ile Asp Leu Asp Leu
355 360 365Ser Thr Val Gln Ala Ser Leu Ser Gly Pro Lys Arg Pro Gln Asp Leu
370 375 380Ile Phe Leu Ser Asp Met Lys Thr Glu Phe Glu Lys Ser Val Thr Ala385 390 395 400Pro Ala Gly Asn Gln Gly His Gly Leu Asp Glu Ser Glu Phe Asp Lys
405 410 415Lys Ala Glu Ile Lys Phe Asn Asp Gly Arg Thr Ser Thr Met Lys Thr
420 425 430Gly Asp Val Ala Ile Ala Ala Ile Thr Ser Cys Thr Asn Thr Ser Asn
435 440 445Pro Tyr Val Met Leu Gly Ala Gly Leu Val Ala Lys Lys Ala Ile Glu
450 455 460Lys Gly Leu Lys Val Pro Asp Tyr Val Lys Thr Ser Leu Ala Pro Gly465 470 475 480Ser Lys Val Val Thr Gly Tyr Leu Arg Asp Ser Gly Leu Gln Glu Tyr
485 490 495Leu Asp Asp Leu Gly Phe Asn Leu Val Gly Tyr Gly Cys Thr Thr Cys
500 505 510Ile Gly Asn Ser Gly Pro Leu Leu Pro Glu Ile Glu Lys Ala Val Ala
515 520 525Asp Glu Asp Leu Leu Val Thr Ser Val Leu Ser Gly Asn Arg Asn Phe
530 535 540Glu Gly Arg Ile His Pro Leu Val Lys Ala Asn Tyr Leu Ala Ser Pro545 550 555 560Gln Leu Val Val Ala Tyr Ala Leu Ala Gly Thr Val Asp Ile Asp Leu
565 570 575His Asn Glu Pro Ile Gly Lys Gly Lys Asp Gly Glu Asp Val Tyr Leu
580 585 590Lys Asp Ile Trp Pro Ser Ile Lys Glu Val Ala Asp Thr Val Asp Ser
595 600 605Val Val Thr Pro Glu Leu Phe Leu Glu Glu Tyr Ala Asn Val Tyr Glu
610 615 620Asn Asn Glu Met Trp Asn Glu Ile Asp Val Thr Asp Ala Pro Leu Tyr625 630 635 640Asp Phe Asp Pro Asn Ser Thr Tyr Ile Gln Asn Pro Ser Phe Phe Gln
645 650 655Gly Leu Ser Lys Glu Pro Gly Thr Ile Glu Pro Leu Lys Asp Leu Arg
660 665 670Ile Met Gly Lys Phe Gly Asp Ser Val Thr Thr Asp His Ile Ser Pro
675 680 685Ala Gly Ala Ile Gly Lys Asp Thr Pro Ala Gly Lys Tyr Leu Leu Asp
690 695 700His Asp Val Pro Ile Arg Glu Phe Asn Ser Tyr Gly Ser Arg Arg Gly705 710 715 720Asn His Glu Val Met Val Arg Gly Thr Phe Ala Asn Ile Arg Ile Lys
725 730 735Asn Gln Leu Ala Pro Gly Thr Glu Gly Gly Phe Thr Thr Tyr Trp Pro
740 745 750Thr Glu Glu Ile Met Pro Ile Tyr Asp Ala Ala Met Arg Tyr Lys Glu
755 760 765Asn Gly Thr Gly Leu Ala Val Leu Ala Gly Asn Asp Tyr Gly Met Gly
770775 780Ser Ser Arg Asp Trp Ala Ala Lys Gly Thr Asn Leu Leu Gly Val Lys785 790 795 800Thr Val Ile Ala Gln Ser Tyr Glu Arg Ile His Arg Ser Asn Leu Val
805 810 815Met Met Gly Val Leu Pro Leu Gln Phe Lys Gln Gly Glu Ser Ala Asp
820 825 830Ser Leu Gly Leu Glu Gly Lys Glu Glu Ile Ser Val Asp Ile Asp Glu
835 840 845Asn Val Lys Pro His Asp Leu Val Thr Val His Ala Lys Lys Glu Asn
850 855 860Gly Glu Val Val Asp Phe Glu Ala Met Val Arg Phe Asp Ser Leu Val865 870 875 880Glu Leu Asp Tyr Tyr Arg His Gly Gly Ile Leu Gln Met Val Leu Arg
885 890 895Asn Lys Leu Ala Gln
900<210>29<211>3006<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(328)..(2514)<400>29gtcgacgacg aaccccccac cgccgaacca gccgccgatc tggtgtggga gacacccggg 60ttctcctccc tgggtgaaca ggtgccacaa ccccgtccca acaggcacac ctaccactgg 120atcgccgggg agagcagcat ggtcacacgc ctgcggcgtg ccctggtgaa ggatcacggc 180ctggacagat cgcaggtggc attcatgggt tattggaggc agggagtggc catgaggggt 240tgatatcgct tccctgaggg tccgcaggcg tgcctcaccc tgtattcttg atagttgaac 300aaaagagccc acataacaag gagactc atg gct aag atc atc tgg acc cgc acc 354
Met Ala Lys Ile Ile Trp Thr Arg Thr
1 5gac gaa gca ccg ctg ctc gcg acc tac tcg ctg aag ccg gtc gtc gag 402Asp Glu Ala Pro Leu Leu Ala Thr Tyr Ser Leu Lys Pro Val Val Glu 10 15 20 25gct ttc gcc gcc acc gcg ggc atc gag gtg gag acc cgc gat atc tct 450Ala Phe Ala Ala Thr Ala Gly Ile Glu Val Glu Thr Arg Asp Ile Ser
30 35 40ctc gcc ggt cgc atc ctc gca cag ttc gcg gac cag ctc ccc gag gag 498Leu Ala Gly Arg Ile Leu Ala Gln Phe Ala Asp Gln Leu Pro Glu Glu
45 50 55cag aag gtc tcc gac gcc ctc gcc gag ctc ggc gaa ctg gct aag acc 546Gln Lys Val Ser Asp Ala Leu Ala Glu Leu Gly Glu Leu Ala Lys Thr
60 65 70ccc gaa gcc aac atc atc aag ctt ccc aac atc tcc gca tcc gta ccg 594Pro Glu Ala Asn Ile Ile Lys Leu Pro Asn Ile Ser Ala Ser Val Pro
75 80 85cag ctc aag gct gcc gta aag gaa ctg cag gaa cag ggc tac gac ctg 642Gln Leu Lys Ala Ala Val Lys Glu Leu Gln Glu Gln Gly Tyr Asp Leu 90 95 100 105ccc gag tac gag gat gcc aag gac cgc tac gcc gct gtc atc ggc tcc 690Pro Glu Tyr Glu Asp Ala Lys Asp Arg Tyr Ala Ala Val Ile Gly Ser
110 115 120aac gtc aac ccg gtc ctg cgc gag ggc aac tcc gac cgc cgc gca ccg 738Asn Val Asn Pro Val Leu Arg Glu Gly Asn Ser Asp Arg Arg Ala Pro
125 130 135gtg gcc gtg aag aac ttc gtg aag aag ttc ccc cac cgc atg ggc gag 786Val Ala Val Lys Asn Phe Val Lys Lys Phe Pro His Arg Met Gly Glu
140 145 150tgg tcc gcc gac tcc aag acc aac gtt gcc acc atg ggt gcc gac gac 834Trp Ser Ala Asp Ser Lys Thr Asn Val Ala Thr Met Gly Ala Asp Asp
155 160 165ttc cgc agc aat gag aag tcc gtg atc atg gac gag gcc gac acc gtg 882Phe Arg Ser Asn Glu Lys Ser Val Ile Met Asp Glu Ala Asp Thr Val170 175 180 185gtg atc aag cat gtc gcc gcc gac ggc acc gag acc gtg ctc aag gac 930Val Ile Lys His Val Ala Ala Asp Gly Thr Glu Thr Val Leu Lys Asp
190 195 200agc ctc ccc ctg ctc aag ggt gag gtc atc gac ggc acc ttc atc tcc 978Ser Leu Pro Leu Leu Lys Gly Glu Val Ile Asp Gly Thr Phe Ile Ser
205 210 215gcc aag gca ctg gac gcc ttc ctg ctc gac cag gtc aaa cgc gcc aag 1026Ala Lys Ala Leu Asp Ala Phe Leu Leu Asp Gln Val Lys Arg Ala Lys
220 225 230gag gag ggc atc ctc ttc tcc gcc cac atg aag gcc acc atg atg aag 1074Glu Glu Gly Ile Leu Phe Ser Ala His Met Lys Ala Thr Met Met Lys
235 240 245gtc tcc gac ccg atc atc ttc ggc cac atc gtc cgc gcc tac ttc gcc 1122Val Ser Asp Pro Ile Ile Phe Gly His Ile Val Arg Ala Tyr Phe Ala250 255 260 265gat gtc tac gca cag tac ggt gag cag ctg ctc gcc gcc ggc ctc aac 1170Asp Val Tyr Ala Gln Tyr Gly Glu Gln Leu Leu Ala Ala Gly Leu Asn
270 275 280ggt gag aac ggt ctc gcc gcc atc tac gcc ggc ctg gac aag ctg gac 1218Gly Glu Asn Gly Leu Ala Ala Ile Tyr Ala Gly Leu Asp Lys Leu Asp
285 290 295aac ggt gcc gag atc aag gca gcc ttc gac aag ggc ctg gaa gag ggc 1266Asn Gly Ala Glu Ile Lys Ala Ala Phe Asp Lys Gly Leu Glu Glu Gly
300 305 310ccc gac ctg gcc atg gtg aac tcc gcc aag ggc atc acc aac ctg cat 1314Pro Asp Leu Ala Met Val Asn Ser Ala Lys Gly Ile Thr Asn Leu His
315 320 325gtg ccc tcc gat gtc atc atc gac gcc tcc atg ccc gcc atg atc cgc 1362Val Pro Ser Asp Val Ile Ile Asp Ala Ser Met Pro Ala Met Ile Arg330 335 340 345acc tcc ggc aag atg tgg aac aag gac gac cag acc cag gat gcc ctg 1410Thr Ser Gly Lys Met Trp Asn Lys Asp Asp Gln Thr Gln Asp Ala Leu
350 355 360gct gtc atc ccg gac tcc tcc tac gcc ggt gtc tac cag acc gtc atc 1458Ala Val Ile Pro Asp Ser Ser Tyr Ala Gly Val Tyr Gln Thr Val Ile
365 370 375gag gac tgc cgc aag aat ggc gcc ttc gat ccg acc acc atg ggc acc 1506Glu Asp Cys Arg Lys Asn Gly Ala Phe Asp Pro Thr Thr Met Gly Thr
380 385 390gtc ccc aac gtc ggt ctg atg gca cag aag gcc gag gag tac ggc tcc 1554Val Pro Asn Val Gly Leu Met Ala Gln Lys Ala Glu Glu Tyr Gly Ser
395 400 405cac gac aag acc ttc cgt atc gag gcc gac ggc aag gta cag gtc gtc 1602His Asp Lys Thr Phe Arg Ile Glu Ala Asp Gly Lys Val Gln Val Val410 415 420 425gcc tcc aac ggt gat gtc ctc atc gag cac gac gtg gag aag ggc gac 1650Ala Ser Asn Gly Asp Val Leu Ile Glu His Asp Val Glu Lys Gly Asp
430 435 440atc tgg cgc gcc tgc cag acc aag gac gcc ccg atc cag gac tgg gtc 1698Ile Trp Arg Ala Cys Gln Thr Lys Asp Ala Pro Ile Gln Asp Trp Val
445 450 455aag ctg gct gtc aac cgc gca cgt ctc tcc ggc atg ccc gct gtg ttc 1746Lys Leu Ala Val Asn Arg Ala Arg Leu Ser Gly Met Pro Ala Val Phe
460 465 470tgg ctg gat ccc gcc cgc gca cac gac cgc aac ctg acc aca ctg gtg 1794Trp Leu Asp Pro Ala Arg Ala His Asp Arg Asn Leu Thr Thr Leu Val
475 480 485gag aag tac ctg gca gac cac gac acc gag ggc ctg gac atc cag atc 1842Glu Lys Tyr Leu Ala Asp His Asp Thr Glu Gly Leu Asp Ile Gln Ile490 495 500 505ctc tcc ccc gtc gag gcc acc cag cac gcc atc gac cgc atc cgc cgc 1890Leu Ser Pro Val Glu Ala Thr Gln His Ala Ile Asp Arg Ile Arg Arg
510 515 520ggc gag gac acc atc tcc gtc acc ggt aac gtc ctg cgt gac tac aac 1938Gly Glu Asp Thr Ile Ser Val Thr Gly Asn Val Leu Arg Asp Tyr Asn
525 530 535acc gac ctc ttc ccg atc ctc gag ctg ggc acc tcc gcc aag atg ctc 1986Thr Asp Leu Phe Pro Ile Leu Glu Leu Gly Thr Ser Ala Lys Met Leu
540 545 550tcc gtc gtg cca ctg atg gcc ggc ggt gga ctc ttc gag acc ggt gcc 2034Ser Val Val Pro Leu Met Ala Gly Gly Gly Leu Phe Glu Thr Gly Ala
555 560 565ggt ggc tcc gcc ccg aag cac gtc cag cag gtc atc gag gaa aac cac 2082Gly Gly Ser Ala Pro Lys His Val Gln Gln Val Ile Glu Glu Asn His570 575 580 585ctg cgc tgg gat tcc ctc ggt gag ttc ctg gcc ctg gcc gag tcc ttc 2130Leu Arg Trp Asp Ser Leu Gly Glu Phe Leu Ala Leu Ala Glu Ser Phe
590 595 600cgc cac gag ctc aac acc cgc aac aac acc aag gcc ggt gtc ctc gcc 2178Arg His Glu Leu Asn Thr Arg Asn Asn Thr Lys Ala Gly Val Leu Ala
605 610 615gat gcc ctg gac cgt gcg acc gag aag ctc ctc aac gag gag aag tcc 2226Asp Ala Leu Asp Arg Ala Thr Glu Lys Leu Leu Asn Glu Glu Lys Ser
620 625 630ccg tcc cgc aag gtc ggc gag atc gac aac cgt ggt tcc cac ttc tgg 2274Pro Ser Arg Lys Val Gly Glu Ile Asp Asn Arg Gly Ser His Phe Trp
635 640 645ctg gcc acc tac tgg gcc gat gaa ctg gcc aac cag acc gag gac gcc 2322Leu Ala Thr Tyr Trp Ala Asp Glu Leu Ala Asn Gln Thr Glu Asp Ala650 655 660 665gag ctg gct gag acc ttc gcc cct gtc gcc gag gcc ctg aac aac cag 2370Glu Leu Ala Glu Thr Phe Ala Pro Val Ala Glu Ala Leu Asn Asn Gln
670 675 680gct gcc gac atc gac gca gca ctc atc ggt gag cag ggc aag cct gtc 2418Ala Ala Asp Ile Asp Ala Ala Leu Ile Gly Glu Gln Gly Lys Pro Val
685 690 695gac ctg ggt ggc tac tac gca ccc tcc gat gag aag acc tcc gcg atc 2466Asp Leu Gly Gly Tyr Tyr Ala Pro Ser Asp Glu Lys Thr Ser Ala Ile
700 705 710atg cgc ccg gtg gcc gca ttc aac gag atc atc gac tcc ctg aag aag 2514Met Arg Pro Val Ala Ala Phe Asn Glu Ile Ile Asp Ser Leu Lys Lys
715 720 725taaccccttc tccggagccg acagccgacg gccacgctcc cccgcccacg ggggatcgtg 2574gccgtcggcc gtttctggca ctggagtgaa cacttcggtg ataatggtga gatgaacagc 2634ccccgtgtcc ccgccatcct gtccgccgtt tccgccgtgg gtctgatcgc tgcgctgggc 2694acccccgttg ccgtcgcaga caccatcacc gcggacaccg accgggaaac ctgcgtggcc 2754agccagaatg acaactccag cgtgatcagg ttctgggatg acctggaggc cgatgtccgt 2814gagcagcgcc tgaccgaact ggatgcacag gaccccggcc tcaagaacga catcgaggcc 2874ttcatcgccg aggacccggt agccccctcc gcagccgatc tccagagacg gctggatgca 2934aatgacgccg gtgagggcct ggccatgctg ctacctgaat cccgcaccga ccccgaggtg 2994gtggacctgc ag 3006<210>30<211>729<212>PRT<213>Corynebacterium thermoaminogenes<400>30Met Ala Lys Ile Ile Trp Thr Arg Thr Asp Glu Ala Pro Leu Leu Ala 1 5 10 15Thr Tyr Ser Leu Lys Pro Val Val Glu Ala Phe Ala Ala Thr Ala Gly
20 25 30Ile Glu Val Glu Thr Arg Asp Ile Ser Leu Ala Gly Arg Ile Leu Ala
35 40 45Gln Phe Ala Asp Gln Leu Pro Glu Glu Gln Lys Val Ser Asp Ala Leu
50 55 60Ala Glu Leu Gly Glu Leu Ala Lys Thr Pro Glu Ala Asn Ile Ile Lys 65 70 75 80Leu Pro Asn Ile Ser Ala Ser Val Pro Gln Leu Lys Ala Ala Val Lys
85 90 95Glu Leu Gln Glu Gln Gly Tyr Asp Leu Pro Glu Tyr Glu Asp Ala Lys
100 105 110Asp Arg Tyr Ala Ala Val Ile Gly Ser Asn Val Asn Pro Val Leu Arg
115 120 125Glu Gly Asn Ser Asp Arg Arg Ala Pro Val Ala Val Lys Asn Phe Val
130 135 140Lys Lys Phe Pro His Arg Met Gly Glu Trp Ser Ala Asp Ser Lys Thr145 150 155 160Asn Val Ala Thr Met Gly Ala Asp Asp Phe Arg Ser Asn Glu Lys Ser
165 170 175Val Ile Met Asp Glu Ala Asp Thr Val Val Ile Lys His Val Ala Ala
180 185 190Asp Gly Thr Glu Thr Val Leu Lys Asp Ser Leu Pro Leu Leu Lys Gly
195 200 205Glu Val Ile Asp Gly Thr Phe Ile Ser Ala Lys Ala Leu Asp Ala Phe
210 215 220Leu Leu Asp Gln Val Lys Arg Ala Lys Glu Glu Gly Ile Leu Phe Ser225 230 235 240Ala His Met Lys Ala Thr Met Met Lys Val Ser Asp Pro Ile Ile Phe
245 250 255Gly His Ile Val Arg Ala Tyr Phe Ala Asp Val Tyr Ala Gln Tyr Gly
260 265 270Glu Gln Leu Leu Ala Ala Gly Leu Asn Gly Glu Asn Gly Leu Ala Ala
275 280 285Ile Tyr Ala Gly Leu Asp Lys Leu Asp Asn Gly Ala Glu Ile Lys Ala
290 295 300Ala Phe Asp Lys Gly Leu Glu Glu Gly Pro Asp Leu Ala Met Val Asn305 310 315 320Ser Ala Lys Gly Ile Thr Asn Leu His Val Pro Ser Asp Val Ile Ile
325 330 335Asp Ala Ser Met Pro Ala Met Ile Arg Thr Ser Gly Lys Met Trp Asn
340 345 350Lys Asp Asp Gln Thr Gln Asp Ala Leu Ala Val Ile Pro Asp Ser Ser
355 360 365Tyr Ala Gly Val Tyr Gln Thr Val Ile Glu Asp Cys Arg Lys Asn Gly
370 375 380Ala Phe Asp Pro Thr Thr Met Gly Thr Val Pro Asn Val Gly Leu Met385 390 395 400Ala Gln Lys Ala Glu Glu Tyr Gly Ser His Asp Lys Thr Phe Arg Ile
405 410 415Glu Ala Asp Gly Lys Val Gln Val Val Ala Ser Asn Gly Asp Val Leu
420 425 430Ile Glu His Asp Val Glu Lys Gly Asp Ile Trp Arg Ala Cys Gln Thr
435 440 445Lys Asp Ala Pro Ile Gln Asp Trp Val Lys Leu Ala Val Asn Arg Ala
450 455 460Arg Leu Ser Gly Met Pro Ala Val Phe Trp Leu Asp Pro Ala Arg Ala465 470 475 480His Asp Arg Asn Leu Thr Thr Leu Val Glu Lys Tyr Leu Ala Asp His
485 490 495Asp Thr Glu Gly Leu Asp Ile Gln Ile Leu Ser Pro Val Glu Ala Thr
500 505 510Gln His Ala Ile Asp Arg Ile Arg Arg Gly Glu Asp Thr Ile Ser Val
515 520 525Thr Gly Asn Val Leu Arg Asp Tyr Asn Thr Asp Leu Phe Pro Ile Leu
530 535 540Glu Leu Gly Thr Ser Ala Lys Met Leu Ser Val Val Pro Leu Met Ala545 550 555 560Gly Gly Gly Leu Phe Glu Thr Gly Ala Gly Gly Ser Ala Pro Lys His
565 570 575Val Gln Gln Val Ile Glu Glu Asn His Leu Arg Trp Asp Ser Leu Gly
580 585 590Glu Phe Leu Ala Leu Ala Glu Ser Phe Arg His Glu Leu Asn Thr Arg
595 600 605Asn Asn Thr Lys Ala Gly Val Leu Ala Asp Ala Leu Asp Arg Ala Thr
610 615 620Glu Lys Leu Leu Asn Glu Glu Lys Ser Pro Ser Arg Lys Val Gly Glu625 630 635 640Ile Asp Asn Arg Gly Ser His Phe Trp Leu Ala Thr Tyr Trp Ala Asp
645 650 655Glu Leu Ala Asn Gln Thr Glu Asp Ala Glu Leu Ala Glu Thr Phe Ala
660 665 670Pro Val Ala Glu Ala Leu Asn Asn Gln Ala Ala Asp Ile Asp Ala Ala
675 680 685Leu Ile Gly Glu Gln Gly Lys Pro Val Asp Leu Gly Gly Tyr Tyr Ala
690 695 700Pro Ser Asp Glu Lys Thr Ser Ala Ile Met Arg Pro Val Ala Ala Phe705 710 715 720Asn Glu Ile Ile Asp Ser Leu Lys Lys
725<210>31<211>2322<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(806)..(2212)<400>31ggtaccccca cgtaccctag gccatcacag caatttttac atcggatatt ttaggtgtgc 60tcataacgtc cttatgaatt tcgcagttat tagttattta aatagagaat caaactccga 120cctagcctct gccgatgcta aaagtcagct gaccccttgg ggcgcttcat ttgaaactgc 180gaccaagctc atgaatgcgc gaaagcattt ccattataag ggtaagctgt aagaatagtg 240ggagaaaatg ttcagtcgtg ttctaactca cttgagaaat tccatttttc tgggcttctc 300tcaaatagat taagtggccc gtatgctgga tttctagaat atttagaagc gcgccaactc 360atgattatgt attgtataag cctcaaagac cgaatagatt actaaeattt aagtggacca 420gagcgttaga agctttgtag agtgctcatt ccttgctgac ggcaagggtt tcctaccatg 480agatagatcg gcagatagtt ggtttgtaaa aatttttaag gacggtccgc aatgtcaatt 540cttgaacaga tcatcttctt catcaacacc atcttgggtt atggtctgca cgctggttct 600tccgcttcca gcaacctttc tcacacgatc ggcctgttct aggcctaatt ggtaataagg 660ctgtgtaaca gtcgcccgcg tgattgtgtc tttttaggcg cccgcgcggg cgattttcgg 720ttttcatctt ttttaaattg agtttggaag atcaagtgcc cccggatgca cgacaatgct 780atgccgaaca cgtattgttg aaatc gtg act gaa cat tat gac gta gta gta 832
Val Thr Glu His Tyr Asp Val Val Val
1 5ctc gga gct ggc ccc ggt ggc tat gtc tcc gcc atc cgc gcc gcg cag 880Leu Gly Ala Gly Pro Gly Gly Tyr Val Ser Ala Ile Arg Ala Ala Gln 10 15 20 25ctc ggt aag aaa gtt gcg gtt atc gag aag cag tac tgg gga ggt gtc 928Leu Gly Lys Lys Val Ala Val Ile Glu Lys Gln Tyr Trp Gly Gly Val
30 35 40tgc ctg aat gtg ggt tgt atc cca tct aag gcg ttg atc aag aac gct 976Cys Leu Asn Val Gly Cys Ile Pro Ser Lys Ala Leu Ile Lys Asn Ala
45 50 55gag atc gcc cac atc ttc aac cat gag aag aag acc ttc ggc atc aac 1024Glu Ile Ala His Ile Phe Asn His Glu Lys Lys Thr Phe Gly Ile Asn
60 65 70ggc gag gtc acc ttc aac tac gag gat gcc cac aag cgt tcc cgt ggt 1072Gly Glu Val Thr Phe Asn Tyr Glu Asp Ala His Lys Arg Ser Arg Gly
75 80 85gtc tcc gac aag atc gtc ggc ggt gtt cac tac ttg atg aag aag aac 1120Val Ser Asp Lys Ile Val Gly Gly Val His Tyr Leu Met Lys Lys Asn 90 95 100 105aag atc acc gag atc gac ggt ttc ggc acc ttc aag gat gcc aag acc 1168Lys Ile Thr Glu Ile Asp Gly Phe Gly Thr Phe Lys Asp Ala Lys Thr
110 115 120atc gag gtg acc gat ggt aag gat gcc ggc aag acc gtc acc ttc gat 1216Ile Glu Val Thr Asp Gly Lys Asp Ala Gly Lys Thr Val Thr Phe Asp
125 130 135gac tgc atc atc gcc acc ggt tcc gtg gtc aac tcc ctc cgt ggt gtt 1264Asp Cys Ile Ile Ala Thr Gly Ser Val Val Asn Ser Leu Arg Gly Val
140 145 150gag ttc tcc gag aac gtg gtc tcc tac gag gag cag atc ctc aac ccg 1312Glu Phe Ser Glu Asn Val Val Ser Tyr Glu Glu Gln Ile Leu Asn Pro
155 160 165gtg gcg cct aag aag atg gtc atc gtc ggt ggc ggc gcc atc ggt atg 1360Val Ala Pro Lys Lys Met Val Ile Val Gly Gly Gly Ala Ile Gly Met170 175 180 185gaa ttc gcc tac gtt ctg ggc aac tac ggt gtg gac gta acc ctc atc 1408Glu Phe Ala Tyr Val Leu Gly Asn Tyr Gly Val Asp Val Thr Leu Ile
190 195 200gag ttc atg gac cgc gtt ctg ccg aac gag gat cca gag gtg tcc aag 1456Glu Phe Met Asp Arg Val Leu Pro Asn Glu Asp Pro Glu Val Ser Lys
205 210 215gtt atc gcc aag gcc tac aag aag atg ggc atc aag ctc ctc ccg ggc 1504Val Ile Ala Lys Ala Tyr Lys Lys Met Gly Ile Lys Leu Leu Pro Gly
220 225 230cac gca acc acc gcg gtg cgc gac aat ggc gat tcc gtt gag gtc gat 1552His Ala Thr Thr Ala Val Arg Asp Asn Gly Asp Ser Val Glu Val Asp
235 240 245tac cag aag aag ggc tcg gac aag acc gag acc atc acc gtc gac cgt 1600Tyr Gln Lys Lys Gly Ser Asp Lys Thr Glu Thr Ile Thr Val Asp Arg250 255 260 265gtt ctt atc tcc gtc ggc ttc cgc cca cgc gtc gag ggc ttc ggc ctg 1648Val Leu Ile Ser Val Gly Phe Arg Pro Arg Val Glu Gly Phe Gly Leu
270 275 280gag aac acc ggc gtc aag ctc acc gaa cgc ggt gcc atc gac att gat 1696Glu Asn Thr Gly Val Lys Leu Thr Glu Arg Gly Ala Ile Asp Ile Asp
285 290 295gag cat atg cgc acc aac gtc gac ggc atc tac gcc atc ggt gac gtc 1744Glu His Met Arg Thr Asn Val Asp Gly Ile Tyr Ala Ile Gly Asp Val
300 305 310acc gcc aag ctg cag ctg gca cac gtc gcc gag gca cag ggc att gtc 1792Thr Ala Lys Leu Gln Leu Ala His Val Ala Glu Ala Gln Gly Ile Val
315 320 325gcc gcc gag aca ctc gcc ggc gca gaa acc cag acc ctg ggc gac tac 1840Ala Ala Glu Thr Leu Ala Gly Ala Glu Thr Gln Thr Leu Gly Asp Tyr330 335 340 345atg atg atg ccg cgt gcc acc ttc tgc aac cca cag gtt gcc tcc ttc 1888Met Met Met Pro Arg Ala Thr Phe Cys Asn Pro Gln Val Ala Ser Phe
350 355 360ggt tac acc gag gag cag gcc aag gag aag tgg ccg gat cga gag atc 1936Gly Tyr Thr Glu Glu Gln Ala Lys Glu Lys Trp Pro Asp Arg Glu Ile
365 370 375aag gtg tcc tcc ttc ccg ttc tcc gcg aac ggc aag gcc gtc ggc ctg 1984Lys Val Ser Ser Phe Pro Phe Ser Ala Asn Gly Lys Ala Val Gly Leu
380 385 390gct gag acc gat ggt ttc gcc aag atc gtc gcc gac gct gag ttc ggt 2032Ala Glu Thr Asp Gly Phe Ala Lys Ile Val Ala Asp Ala Glu Phe Gly
395 400 405gaa ctg ctg ggt ggc cac att gtc ggt gcc aac gcc tcc gag ctg ctc 2080Glu Leu Leu Gly Gly His Ile Val Gly Ala Asn Ala Ser Glu Leu Leu410 415 420 425aac gag ctg gtg ctg gcc cag aac tgg gat ctc acc acc gag gag atc 2128Asn Glu Leu Val Leu Ala Gln Asn Trp Asp Leu Thr Thr Glu Glu Ile
430 435 440agc cgc agc gtc cac atc cac ccg acc ctg tcg gag gct gtc aag gaa 2176Ser Arg Ser Val His Ile His Pro Thr Leu Ser Glu Ala Val Lys Glu
445 450 455gct gcc cac ggc gtc aac ggc cac atg atc aac ttc taaatcccgt2222Ala Ala His Gly Val Asn Gly His Met Ile Asn Phe
460 465cagacaaatg caaatcccct caccgatggc atatcggtga ggggattttc tcatgcacgt 2282aaaatcataa tccatggcaa ggaaagtcga caacagcgcc 2322<210>32<211>469<212>PRT<213>Corynebacterium thermoaminogenes<400>32Val Thr Glu His Tyr Asp Val Val Val Leu Gly Ala Gly Pro Gly Gly 1 5 10 15Tyr Val Ser Ala Ile Arg Ala Ala Gln Leu Gly Lys Lys Val Ala Val
20 25 30Ile Glu Lys Gln Tyr Trp Gly Gly Val Cys Leu Asn Val Gly Cys Ile
35 40 45Pro Ser Lys Ala Leu Ile Lys Asn Ala Glu Ile Ala His Ile Phe Asn
50 55 60His Glu Lys Lys Thr Phe Gly Ile Asn Gly Glu Val Thr Phe Asn Tyr 65 70 75 80Glu Asp Ala His Lys Arg Ser Arg Gly Val Ser Asp Lys Ile Val Gly
85 90 95Gly Val His Tyr Leu Met Lys Lys Asn Lys Ile Thr Glu Ile Asp Gly
100 105 110Phe Gly Thr Phe Lys Asp Ala Lys Thr Ile Glu Val Thr Asp Gly Lys
115 120 125Asp Ala Gly Lys Thr Val Thr Phe Asp Asp Cys Ile Ile Ala Thr Gly
130 135 140Ser Val Val Asn Ser Leu Arg Gly Val Glu Phe Ser Glu Asn Val Val145 150 155 160Ser Tyr Glu Glu Gln Ile Leu Asn Pro Val Ala Pro Lys Lys Met Val
165 170 175Ile Val Gly Gly Gly Ala Ile Gly Met Glu Phe Ala Tyr Val Leu Gly
180 185 190Asn Tyr Gly Val Asp Val Thr Leu Ile Glu Phe Met Asp Arg Val Leu
195 200 205Pro Asn Glu Asp Pro Glu Val Ser Lys Val Ile Ala Lys Ala Tyr Lys
210 215 220Lys Met Gly Ile Lys Leu Leu Pro Gly His Ala Thr Thr Ala Val Arg225 230 235 240Asp Asn Gly Asp Ser Val Glu Val Asp Tyr Gln Lys Lys Gly Ser Asp
245 250 255Lys Thr Glu Thr Ile Thr Val Asp Arg Val Leu Ile Ser Val Gly Phe
260 265 270Arg Pro Arg Val Glu Gly Phe Gly Leu Glu Asn Thr Gly Val Lys Leu
275 280 285Thr Glu Arg Gly Ala Ile Asp Ile Asp Glu His Met Arg Thr Asn Val
290 295 300Asp Gly Ile Tyr Ala Ile Gly Asp Val Thr Ala Lys Leu Gln Leu Ala305 310 315 320His Val Ala Glu Ala Gln Gly Ile Val Ala Ala Glu Thr Leu Ala Gly
325 330 335Ala Glu Thr Gln Thr Leu Gly Asp Tyr Met Met Met Pro Arg Ala Thr
340 345 350Phe Cys Asn Pro Gln ValAla Ser Phe Gly Tyr Thr Glu Glu Gln Ala
355360 365Lys Glu Lys Trp Pro Asp Arg Glu Ile Lys Val Ser Ser Phe Pro Phe
370 375 380Ser Ala Asn Gly Lys Ala Val Gly Leu Ala Glu Thr Asp Gly Phe Ala385 390 395 400Lys Ile Val Ala Asp Ala Glu Phe Gly Glu Leu Leu Gly Gly His Ile
405 410 415Val Gly Ala Asn Ala Ser Glu Leu Leu Asn Glu Leu Val Leu Ala Gln
420 425 430Asn Trp Asp Leu Thr Thr Glu Glu Ile Ser Arg Ser Val His Ile His
435 440 445Pro Thr Leu Ser Glu Ala Val Lys Glu Ala Ala His Gly Val Asn Gly
450 455 460His Met Ile Asn Phe465<210>33<211>4096<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(250)..(3951)<400>33ccggatcatc gtggttgacg ggggacgtat catcgaggat ggttcccacg atgaacttct 60gggagcgaat ggaacctacg caacaatgtg gcatttagta gggtgacagg atattttagg 120aaagacttgt taccaaaagg tgctaatact ggggtgctag gtccccgcga ccggaaccag 180cgttacagtg gataaaataa agcccattta gaaccctcaa caagcaagga aaagaggcga 240gtacctgcc gtg agc agc gct agt act ttc ggc cag aac gcg tgg ctg gtg 291
Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp Leu Val
1 5 10gat gag atg ttc cag cag ttc aag aag gac ccc cag tcc gtg gac aag 339Asp Glu Met Phe Gln Gln Phe Lys Lys Asp Pro Gln Ser Val Asp Lys 15 20 25 30gaa tgg aga gag ctc ttc gag tct cag ggg ggt ccc cag gct gaa aag 387Glu Trp Arg Glu Leu Phe Glu Ser Gln Gly Gly Pro Gln Ala Glu Lys
35 40 45gct acc ccc gcc acc ccc gaa gcc aag aag gca gct tcg tcg cag tcc 435Ala Thr Pro Ala Thr Pro Glu Ala Lys Lys Ala Ala Ser Ser Gln Ser
50 55 60tca act tcc gga cag tcc acc gcc aag gct gcc cct gcc gcc aag acc 483Ser Thr Ser Gly Gln Ser Thr Ala Lys Ala Ala Pro Ala Ala Lys Thr
65 70 75gca ccg gcc tct gcg cca gcc aag gct gcc cct gtt aag caa aac cag 531Ala Pro Ala Ser Ala Pro Ala Lys Ala Ala Pro Val Lys Gln Asn Gln
80 85 90gcg tcc aag cct gcc aag aag gcc aag gag tcc ccc ctg tcc aag cca 579Ala Ser Lys Pro Ala Lys Lys Ala Lys Glu Ser Pro Leu Ser Lys Pro 95 100 105 110gct gcc atg cct gag ccg gga acc acc cca ctc agg ggc atc ttc aag 627Ala Ala Met Pro Glu Pro Gly Thr Thr Pro Leu Arg Gly Ile Phe Lys
115 120 125tcc atc gcc aag aac atg gac ctc tcc ctc gag gtg ccc acc gcc acc 675Ser Ile Ala Lys Asn Met Asp Leu Ser Leu Glu Val Pro Thr Ala Thr
130 135 140tcc gtc cgc gac atg ccc gcg cgc ctc atg ttc gag aac cgc gcc atg 723Ser Val Arg Asp Met Pro Ala Arg Leu Met Phe Glu Asn Arg Ala Met
145 150 155gtc aac gac cag ctc aag cgc acc cgt ggc ggc aag atc tcc ttc acc 771Val Asn Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr
160 165 170cac atc atc ggc tac gcc atg gtg aag gct gtc atg gca cac ccg gac 819His Ile Ile Gly Tyr Ala Met Val Lys Ala Val Met Ala His Pro Asp175 180 185 190atg aac aac tcc tat gac atc gtc gac ggc aag ccg tcc ctg gtc gtc 867Met Asn Asn Ser Tyr Asp Ile Val Asp Gly Lys Pro Ser Leu Val Val
195 200 205ccg gag cac atc aac ctc ggc ctg gcc atc gac ctc ccc cag aag gac 915Pro Glu His Ile Asn Leu Gly Leu Ala Ile Asp Leu Pro Gln Lys Asp
210 215 220ggc tcc cgt gcc ctc gtg gtc gcc gcc atc aag gaa acc gag aag atg 963Gly Ser Arg Ala Leu Val Val Ala Ala Ile Lys Glu Thr Glu Lys Met
225 230 235acc ttc tcc cag ttc ctg gag gcc tat gag gac gtt gtg gca cgc tcc 1011Thr Phe Ser Gln Phe Leu Glu Ala Tyr Glu Asp Val Val Ala Arg Ser
240 245 250cgc gtc ggc aag ctc acc atg gat gac tac cag ggt gtc acc atc tcc 1059Arg Val Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly Val Thr Ile Ser255 260 265 270ttg acc aac ccg ggt ggc atc ggt acc cgc cac tcc atc ccg cgt ctg 1107Leu Thr Asn Pro Gly Gly Ile Gly Thr Arg His Ser Ile Pro Arg Leu
275 280 285acc aag ggc cag ggc acc atc atc ggt gtc ggt tcc atg gac tac ccg 1155Thr Lys Gly Gln Gly Thr Ile Ile Gly Val Gly Ser Met Asp Tyr Pro
290 295 300gcc gag ttc cag ggt gcc tcc gag gac cgt ctc gcc gag ctc ggt gtg 1203Ala Glu Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala Glu Leu Gly Val
305 310 315ggc aag ctc gtc acc atc acc tcc acc tac gat cac cgc gtc atc cag 1251Gly Lys Leu Val Thr Ile Thr Ser Thr Tyr Asp His Arg Val Ile Gln
320 325 330ggc gcg gaa tcc ggt gag ttc ctg cgc acc atg tcc cag ctg ctc gtg 1299Gly Ala Glu Ser Gly Glu Phe Leu Arg Thr Met Ser Gln Leu Leu Val335 340 345 350gac gat gcg ttc tgg gat cac atc ttc gag gag atg aac gtt ccc tac 1347Asp Asp Ala Phe Trp Asp His Ile Phe Glu Glu Met Asn Val Pro Tyr
355 360 365acc ccg atg cgc tgg gca cag gac ctg ccc aac acc ggt gtg gac aag 1395Thr Pro Met Arg Trp Ala Gln Asp Leu Pro Asn Thr Gly Val Asp Lys
370 375 380aac acc cgt gtc atg cag ctc atc gag gcc tac cgc tcc cgc ggt cac 1443Asn Thr Arg Val Met Gln Leu Ile Glu Ala Tyr Arg Ser Arg Gly His
385 390 395ctc atc gcc gac acc aac cca ctg ccc tgg gtc cag ccc ggc atg ccc 1491Leu Ile Ala Asp Thr Asn Pro Leu Pro Trp Val Gln Pro Gly Met Pro
400 405 410gtc ccg gat cac cgt gac ctc gac atc gag acc cac ggc ctg acc ctg 1539Val Pro Asp His Arg Asp Leu Asp Ile Glu Thr His Gly Leu Thr Leu415 420 425 430tgg gat ctg gac cgt acc ttc cac gtc ggt ggt ttc ggt ggc aag gag 1587Trp Asp Leu Asp Arg Thr Phe His Val Gly Gly Phe Gly Gly Lys Glu
435 440 445acc atg acc ctg cgc gag gtg ctc agc cgc ctc cgc gcc gcc tac acc 1635Thr Met Thr Leu Arg Glu Val Leu Ser Arg Leu Arg Ala Ala Tyr Thr
450 455 460ctc aag gtc ggc tcc gag tac acc cac atc ctc gac cgc gat gag cgc 1683Leu Lys Val Gly Ser Glu Tyr Thr His Ile Leu Asp Arg Asp Glu Arg
465 470 475acc tgg ctg cag gac cgc ctc gag gcc ggt atg ccc aag ccc acc gcc 1731Thr Trp Leu Gln Asp Arg Leu Glu Ala Gly Met Pro Lys Pro Thr Ala
480 485 490gcc gag cag aag tac atc ctg cag aag ctc aac gcc gcc gag gca ttc 1779Ala Glu Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala Ala Glu Ala Phe495 500 505 510gag aac ttc ctg cag acc aag tac gtc ggc cag aag cgt ttc tcc ctc 1827Glu Asn Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys Arg Phe Ser Leu
515 520 525gag ggt gcc gag tca ctg atc ccg ctg atg gac tcc gcc atc gac acc 1875Glu Gly Ala Glu Ser Leu Ile Pro Leu Met Asp Ser Ala Ile Asp Thr
530 535 540gcc gca ggc cag ggc ctt gac gag gtc gtc atc ggc atg ccc cac cgt 1923Ala Ala Gly Gln Gly Leu Asp Glu Val Val Ile Gly Met Pro His Arg
545 550 555ggt cgc ctc aac gtg ctg ttc aac atc gtc ggc aag cca ctg gcc tcg 1971Gly Arg Leu Asn Val Leu Phe Asn Ile Val Gly Lys Pro Leu Ala Ser
560 565 570atc ttc aac gag ttc gag ggc cag atg gag cag ggc cag atc ggt ggc 2019Ile Phe Asn Glu Phe Glu Gly Gln Met Glu Gln Gly Gln Ile Gly Gly575 580 585 590tcc ggt gac gtg aag tac cac ctc ggt tcc gag ggc acc cac ctg cag 2067Ser Gly Asp Val Lys Tyr His Leu Gly Ser Glu Gly Thr His Leu Gln
595 600 605atg ttc ggc gac ggc gag atc aag gtc tcc ctc acc gcc aac ccc tcc 2115Met Phe Gly Asp Gly Glu Ile Lys Val Ser Leu Thr Ala Asn Pro Ser
610 615 620cac ctc gag gcc gtc aac ccg gtc gtg gag ggc atc gtc cgc gcc aag 2163His Leu Glu Ala Val Asn Pro Val Val Glu Gly Ile Val Arg Ala Lys
625 630 635cag gac atc ctg gac aag ggc ccg gac ggc tac acc gtc gtc ccg ctg 2211Gln Asp Ile Leu Asp Lys Gly Pro Asp Gly Tyr Thr Val Val Pro Leu
640 645 650ctg ctc cac ggt gac gcc gcc ttc gcc ggc ctg ggc atc gtg ccc gag 2259Leu Leu His Gly Asp Ala Ala Phe Ala Gly Leu Gly Ile Val Pro Glu655 660 665 670acc atc aac ctc gca gcc ctg cgt ggt tac gat gtc ggt ggc acc atc 2307Thr Ile Asn Leu Ala Ala Leu Arg Gly Tyr Asp Val Gly Gly Thr Ile
675 680 685cac atc gtg gtc aac aac cag atc ggc ttc acc acc acc ccg gac tcc 2355His Ile Val Val Asn Asn Gln Ile Gly Phe Thr Thr Thr Pro Asp Ser
690 695 700agc cgt tcc atg cac tac gcc acc gac tgc gcc aag gcc ttc ggt tgc 2403Ser Arg Ser Met His Tyr Ala Thr Asp Cys Ala Lys Ala Phe Gly Cys
705 710 715ccg gtg ttc cac gtc aac ggt gac gac ccc gag gct gtg gtc tgg gtc 2451Pro Val Phe His Val Asn Gly Asp Asp Pro Glu Ala Val Val Trp Val
720 725 730ggc cag ctg gcc acc gag tac cgt cgc cgc ttc ggc aag gat gtc ttc 2499Gly Gln Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly Lys Asp Val Phe735 740 745 750atc gac ctc atc tgc tac cgc ctg cgc ggc cac aac gag gct gat gac 2547Ile Asp Leu Ile Cys Tyr Arg Leu Arg Gly His Asn Glu Ala Asp Asp
755 760 765cca tcc atg acc cag ccg aag atg tac gag ctg atc acc ggc cgc gac 2595Pro Ser Met Thr Gln Pro Lys Met Tyr Glu Leu Ile Thr Gly Arg Asp
770 775 780tcc gtg cgt gcc acc tac acc gag gac ctc ctc ggc cgt ggt gac ctc 2643Ser Val Arg Ala Thr Tyr Thr Glu Asp Leu Leu Gly Arg Gly Asp Leu
785 790 795tcc ccc gag gac gcc gag gcc gtt gtc cgc gac ttc cac gac cag atg 2691Ser Pro Glu Asp Ala Glu Ala Val Val Arg Asp Phe His Asp Gln Met
800 805 810gaa tcc gtg ttc aac gag gtc aag gaa gcc ggc aag aag cag cct gat 2739Glu Ser Val Phe Asn Glu Val Lys Glu Ala Gly Lys Lys Gln Pro Asp815 820 825 830gag cag acc ggc atc acc ggt tcc cag gaa ctg acc cgt ggc ctg gac 2787Glu Gln Thr Gly Ile Thr Gly Ser Gln Glu Leu Thr Arg Gly Leu Asp
835 840 845acc aac atc acc cgc gag gaa ctg gtc gaa ctc ggc cag gcc ttc gtc 2835Thr Asn Ile Thr Arg Glu Glu Leu Val Glu Leu Gly Gln Ala Phe Val
850 855 860aac acc cca gag ggc ttc acc tac cac cca cgt gtg gca ccg gtg gcc 2883Asn Thr Pro Glu Gly Phe Thr Tyr His Pro Arg Val Ala Pro Val Ala
865 870 875aag aag cgt gcc gag tcc gtc acc gag ggt ggc atc gac tgg gca tgg 2931Lys Lys Arg Ala Glu Ser Val Thr Glu Gly Gly Ile Asp Trp Ala Trp
880 885 890ggc gag ctc atc gcc ttc ggc tcc ctg gcc acc tcc ggc agg ctg gtc 2979Gly Glu Leu Ile Ala Phe Gly Ser Leu Ala Thr Ser Gly Arg Leu Val895 900 905 910cgc ctc gcc ggt gag gat tcc cgc cgt ggt acc ttc acc cag cgt cac 3027Arg Leu Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe Thr Gln Arg His
915 920 925gcc gtg gcc atc gac ccg aac acc gcc gag gag ttc aac ccg ctc cac 3075Ala Val Ala Ile Asp Pro Asn Thr Ala Glu Glu Phe Asn Pro Leu His
930 935 940gag ctg gca cag gcc aag ggc ggc ggc aag ttc ctc gtc tac aac tcc 3123Glu Leu Ala Gln Ala Lys Gly Gly Gly Lys Phe Leu Val Tyr Asn Ser
945 950 955gcg ctg acc gag tac gcg ggt atg ggc ttc gaa tac ggc tac tcc gtg 3171Ala Leu Thr Glu Tyr Ala Gly Met Gly Phe Glu Tyr Gly Tyr Ser Val
960 965 970ggc aac ccg gac gcc gtg gtg tcc tgg gag gca cag ttc ggt gac ttc 3219Gly Asn Pro Asp Ala Val Val Ser Trp Glu Ala Gln Phe Gly Asp Phe975 980 985 990gcc aac ggt gca cag acc atc atc gat gag tac atc tcc tcc ggt gag 3267Ala Asn Gly Ala Gln Thr Ile Ile Asp Glu Tyr Ile Ser Ser Gly Glu
99510001005gcc aag tgg ggc cag acc tcc tcg gtc atc ctg ctg ctg ccc cac ggt 3315Ala Lys Trp Gly Gln Thr Ser Ser Val Ile Leu Leu Leu Pro His Gly
101010151020tac gag ggc cag ggt ccg gac cac tcc tcc gca cgc atc gag cgt ttc 3363Tyr Glu Gly Gln Gly Pro Asp His Ser Ser Ala Arg Ile Glu Arg Phe
102510301035ctg cag ctg tgc gcc gag ggt tcc atg acc atc gcc cag ccg acc acc 3411Leu Gln Leu Cys Ala Glu Gly Ser Met Thr Ile Ala Gln Pro Thr Thr 104010451050ccg gcg aac tac ttc cac ctg ctg cgt cgt cac gca ctg ggc aag atg 3459Pro Ala Asn Tyr Phe His Leu Leu Arg Arg His Ala Leu Gly Lys Met1055 106010651070aag cgc ccg ctg gtc gtc ttc acc ccg aag tcc atg ctg cgc aac aag 3507Lys Arg Pro Leu Val Val Phe Thr Pro Lys Ser Met Leu Arg Asn Lys
107510801085gcc gcc acc tcc gct ccg gag gag ttc acc gag gtc acc cgc ttc aag 3555Ala Ala Thr Ser Ala Pro Glu Glu Phe Thr Glu Val Thr Arg Phe Lys
1090 10951100tcc gtg atc gac gat ccg aac gtg gcg gat gcc tcc aag gtg aag aag 3603Ser Val Ile Asp Asp Pro Asn Val Ala Asp Ala Ser Lys Val Lys Lys
110511101115atc atg ctg tgc tcc ggc aag atc tac tac gaa ctg gcc aag cgc aag 3651Ile Met Leu Cys Ser Gly Lys Ile Tyr Tyr Glu Leu Ala Lys Arg Lys 112011251130gag aag gac aac cgc gac gac atc gcg atc gtg cgc atc gag atg ctg 3699Glu Lys Asp Asn Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu1135 114011451150cac ccg atc ccg ttc aac cgt ctg cgc gac gcc ttc gac ggc tac ccc 3747His Pro Ile Pro Phe Asn Arg Leu Arg Asp Ala Phe Asp Gly Tyr Pro
115511601165aac gcc gag gag atc ctg ttc gtt cag gac gag ccg gca aac cag ggt 3795Asn Ala Glu Glu Ile Leu Phe Val Gln Asp Glu Pro Ala Asn Gln Gly
117011751180gcc tgg ccg ttc tac cag gag cac ctg ccc aac ctc atc gag ggc atg 3843Ala Trp Pro Phe Tyr Gln Glu His Leu Pro Asn Leu Ile Glu Gly Met
118511901195ctc ccg atg cgt cgc atc tcg cgc cgt tcc cag tcc tcg act gcg acc 3891Leu Pro Met Arg Arg Ile Ser Arg Arg Ser Gln Ser Ser Thr Ala Thr 120012051210ggt atc gcg aag gtg cac acc atc gag cag cag aag ctg ctg gat gat 3939Gly Ile Ala Lys Val His Thr Ile Glu Gln Gln Lys Leu Leu Asp Asp1215 122012251230gcg ttc aac gca taaacgttaa tacagcggtt gataccttga accccgccgc 3991Ala Phe Asn Alaaccctttaga tgcgggcggg gttttgcttt gcctgcatag gcgataatat tcatatacac 4051ccatcacgtt taagttctgc atttggatcg tgcgagcatc ccggt 4096<210>34<211>1234<212>PRT<213>Corynebacterium thermoaminogenes<400>34Val Ser Ser Ala Ser Thr Phe Gly Gln Asn Ala Trp Leu Val Asp Glu 1 5 10 15Met Phe Gln Gln Phe Lys Lys Asp Pro Gln Ser Val Asp Lys Glu Trp
20 25 30Arg Glu Leu Phe Glu Ser Gln Gly Gly Pro Gln Ala Glu Lys Ala Thr
35 40 45Pro Ala Thr Pro Glu Ala Lys Lys Ala Ala Ser Ser Gln Ser Ser Thr
50 55 60Ser Gly Gln Ser Thr Ala Lys Ala Ala Pro Ala Ala Lys Thr Ala Pro 65 70 75 80Ala Ser Ala Pro Ala Lys Ala Ala Pro Val Lys Gln Asn Gln Ala Ser
85 90 95Lys Pro Ala Lys Lys Ala Lys Glu Ser Pro Leu Ser Lys Pro Ala Ala
100 105 110Met Pro Glu Pro Gly Thr Thr Pro Leu Arg Gly Ile Phe Lys Ser Ile
115 120 125Ala Lys Asn Met Asp Leu Ser Leu Glu Val Pro Thr Ala Thr Ser Val
130 135 140Arg Asp Met Pro Ala Arg Leu Met Phe Glu Asn Arg Ala Met Val Asn145 150 155 160Asp Gln Leu Lys Arg Thr Arg Gly Gly Lys Ile Ser Phe Thr His Ile
165 170 175Ile Gly Tyr Ala Met Val Lys Ala Val Met Ala His Pro Asp Met Asn
180 185 190Asn Ser Tyr Asp Ile Val Asp Gly Lys Pro Ser Leu Val Val Pro Glu
195 200 205His Ile Asn Leu Gly Leu Ala Ile Asp Leu Pro Gln Lys Asp Gly Ser
210 215 220Arg Ala Leu Val Val Ala Ala Ile Lys Glu Thr Glu Lys Met Thr Phe225 230 235 240Ser Gln Phe Leu Glu Ala Tyr Glu Asp Val Val Ala Arg Ser Arg Val
245 250 255Gly Lys Leu Thr Met Asp Asp Tyr Gln Gly Val Thr Ile Ser Leu Thr
260 265 270Asn Pro Gly Gly Ile Gly Thr Arg His Ser Ile Pro Arg Leu Thr Lys
275 280 285Gly Gln Gly Thr Ile Ile Gly Val Gly Ser Met Asp Tyr Pro Ala Glu
290 295 300Phe Gln Gly Ala Ser Glu Asp Arg Leu Ala Glu Leu Gly Val Gly Lys305 310 315 320Leu Val Thr Ile Thr Ser Thr Tyr Asp His Arg Val Ile Gln Gly Ala
325 330 335Glu Ser Gly Glu Phe Leu Arg Thr Met Ser Gln Leu Leu Val Asp Asp
340 345 350Ala Phe Trp Asp His Ile Phe Glu Glu Met Asn Val Pro Tyr Thr Pro
355 360 365Met Arg Trp Ala Gln Asp Leu Pro Asn Thr Gly Val Asp Lys Asn Thr
370 375 380Arg Val Met Gln Leu Ile Glu Ala Tyr Arg Ser Arg Gly His Leu Ile385 390 395 400Ala Asp Thr Asn Pro Leu Pro Trp Val Gln Pro Gly Met Pro Val Pro
405 410 415Asp His Arg Asp Leu Asp Ile Glu Thr His Gly Leu Thr Leu Trp Asp
420 425 430Leu Asp Arg Thr Phe His Val Gly Gly Phe Gly Gly Lys Glu Thr Met
435 440 445Thr Leu Arg Glu Val Leu Ser Arg Leu Arg Ala Ala Tyr Thr Leu Lys
450 455 460Val Gly Ser Glu Tyr Thr His Ile Leu Asp Arg Asp Glu Arg Thr Trp465 470 475 480Leu Gln Asp Arg Leu Glu Ala Gly Met Pro Lys Pro Thr Ala Ala Glu
485 490 495Gln Lys Tyr Ile Leu Gln Lys Leu Asn Ala Ala Glu Ala Phe Glu Asn
500 505 510Phe Leu Gln Thr Lys Tyr Val Gly Gln Lys Arg Phe Ser Leu Glu Gly
515 520 525Ala Glu Ser Leu Ile Pro Leu Met Asp Ser Ala Ile Asp Thr Ala Ala
530 535 540Gly Gln Gly Leu Asp Glu Val Val Ile Gly Met Pro His Arg Gly Arg545 550 555 560Leu Asn Val Leu Phe Asn Ile Val Gly Lys Pro Leu Ala Ser Ile Phe
565 570 575Asn Glu Phe Glu Gly Gln Met Glu Gln Gly Gln Ile Gly Gly Ser Gly
580 585 590Asp Val Lys Tyr His Leu Gly Ser Glu Gly Thr His Leu Gln Met Phe
595 600 605Gly Asp Gly Glu Ile Lys Val Ser Leu Thr Ala Asn Pro Ser His Leu
610 615 620Glu Ala Val Asn Pro Val Val Glu Gly Ile Val Arg Ala Lys Gln Asp625 630 635 640Ile Leu Asp Lys Gly Pro Asp Gly Tyr Thr Val Val Pro Leu Leu Leu
645 650 655His Gly Asp Ala Ala Phe Ala Gly Leu Gly Ile Val Pro Glu Thr Ile
660 665 670Asn Leu Ala Ala Leu Arg Gly Tyr Asp Val Gly Gly Thr Ile His Ile
675 680 685Val Val Asn Asn Gln Ile Gly Phe Thr Thr Thr Pro Asp Ser Ser Arg
690 695 700Ser Met His Tyr Ala Thr Asp Cys Ala Lys Ala Phe Gly Cys Pro Val705 710 715 720Phe His Val Asn Gly Asp Asp Pro Glu Ala Val Val Trp Val Gly Gln
725 730 735Leu Ala Thr Glu Tyr Arg Arg Arg Phe Gly Lys Asp Val Phe Ile Asp
740 745 750Leu Ile Cys Tyr Arg Leu Arg Gly His Asn Glu Ala Asp Asp Pro Ser
755 760765Met Thr Gln Pro Lys Met Tyr Glu Leu Ile Thr Gly Arg Asp Ser Val
770 775 780Arg Ala Thr Tyr Thr Glu Asp Leu Leu Gly Arg Gly Asp Leu Ser Pro785 790 795 800Glu Asp Ala Glu Ala Val Val Arg Asp Phe His Asp Gln Met Glu Ser
805 810 815Val Phe Asn Glu Val Lys Glu Ala Gly Lys Lys Gln Pro Asp Glu Gln
820 825 830Thr Gly Ile Thr Gly Ser Gln Glu Leu Thr Arg Gly Leu Asp Thr Asn
835 840 845Ile Thr Arg Glu Glu Leu Val Glu Leu Gly Gln Ala Phe Val Asn Thr
850 855 860Pro Glu Gly Phe Thr Tyr His Pro Arg Val Ala Pro Val Ala Lys Lys865 870 875 880Arg Ala Glu Ser Val Thr Glu Gly Gly Ile Asp Trp Ala Trp Gly Glu
885 890 895Leu Ile Ala Phe Gly Ser Leu Ala Thr Ser Gly Arg Leu Val Arg Leu
900 905 910Ala Gly Glu Asp Ser Arg Arg Gly Thr Phe Thr Gln Arg His Ala Val
915 920 925Ala Ile Asp Pro Asn Thr Ala Glu Glu Phe Asn Pro Leu His Glu Leu
930 935 940Ala Gln Ala Lys Gly Gly Gly Lys Phe Leu Val Tyr Asn Ser Ala Leu945 950 955 960Thr Glu Tyr Ala Gly Met Gly Phe Glu Tyr Gly Tyr Ser Val Gly Asn
965 970 975Pro Asp Ala Val Val Ser Trp Glu Ala Gln Phe Gly Asp Phe Ala Asn
980 985 990Gly Ala Gln Thr Ile Ile Asp Glu Tyr Ile Ser Ser Gly Glu Ala Lys
99510001005Trp Gly Gln Thr Ser Ser Val Ile Leu Leu Leu Pro His Gly Tyr Glu 101010151020Gly Gln Gly Pro Asp His Ser Ser Ala Arg Ile Glu Arg Phe Leu Gln025103010351040Leu Cys Ala Glu Gly Ser Met Thr Ile Ala Gln Pro Thr Thr Pro Ala
104510501055Asn Tyr Phe His Leu Leu Arg Arg His Ala Leu Gly Lys Met Lys Arg
106010651070Pro Leu Val Val Phe Thr Pro Lys Ser Met Leu Arg Asn Lys Ala Ala
107510801085Thr Ser Ala Pro Glu Glu Phe Thr Glu Val Thr Arg Phe Lys Ser Val 109010951100Ile Asp Asp Pro Asn Val Ala Asp Ala Ser Lys Val Lys Lys Ile Met105111011151120Leu Cys Ser Gly Lys Ile Tyr Tyr Glu Leu Ala Lys Arg Lys Glu Lys
112511301135Asp Asn Arg Asp Asp Ile Ala Ile Val Arg Ile Glu Met Leu His Pro
114011451150Ile Pro Phe Asn Arg Leu Arg Asp Ala Phe Asp Gly Tyr Pro Asn Ala
115511601165Glu Glu Ile Leu Phe Val Gln Asp Glu Pro Ala Asn Gln Gly Ala Trp 117011751180Pro Phe Tyr Gln Glu His Leu Pro Asn Leu Ile Glu Gly Met Leu Pro185119011951200Met Arg Arg Ile Ser Arg Arg Ser Gln Ser Ser Thr Ala Thr Gly Ile
120512101215Ala Lys Val His Thr Ile Glu Gln Gln Lys Leu Leu Asp Asp Ala Phe
1220 12251230Asn Ala<210>35<211>20<212>DNA<213>人工序列<220><223>人工序列的描述aceA的引物<400>35cctctaccca gcgaactccg20<210>36<211>20<212>DNA<213>人工序列<220><223>人工序列的描述aceA的引物<400>36ctgccttgaa ctcacggttc20<210>37<211>20<212>DNA<213>人工序列<220><223>人工序列的描述accBC的引物<400>37catccacccc ggctacggct20<210>38<211>20<212>DNA<213>人工序列<220><223>人工序列的描述accBC的引物<400>38cggtgactgg gtgttccacc20<210>39<211>20<212>DNA<213>人工序列<220><223>人工序列的描述dtsR1的引物<400>39acggcccagc cctgaccgac20<210>40<211>20<212>DNA<213>人工序列<220><223>人工序列的描述dtsR1的引物<400>40agcagcgccc atgacggcga20<210>41<211>20<212>DNA<213>人工序列<220><223>人工序列的描述dtsR2的引物<400>41acggcccagc cctgaccgac20<210>42<211>20<212>DNA<213>人工序列<220><223>人工序列的描述dtsR2的引物<400>42agcagcgccc atgacggcga20<210>43<211>20<212>DNA<213>人工序列<220><223>人工序列的描述pfk的引物<400>43cgtcatccga ggaatcgtcc20<210>44<211>20<212>DNA<213>人工序列<220><223>人工序列的描述pfk的引物<400>44cgtggcggcc catgacctcc21<210>45<211>17<212>DNA<213>人工序列<220><223>人工序列的描述scrB的引物<220><221>不确定<222>(3)<223>n＝a or g or c or t<400>45ggncghytba aygaycc 17<210>46<211>20<212>DNA<213>人工序列<220><223>人工序列的描述scrB的引物<220><221>不确定<222>(18)<223>n＝a or g or c or t<400>46ggrcaytccc acatrtancc20<210>47<211>20<212>DNA<213>人工序列<220><223>人工序列的描述gluABCD的引物<400>47ccatccggat ccggcaagtc20<210>48<211>20<212>DNA<213>人工序列<220><223>人工序列的描述gluABCD的引物<400>48aatcccatct cgtgggtaac20<210>49<211>23<212>DNA<213>人工序列<220><223>人工序列的描述pdhA的引物<400>49actgtgtcca tgggtcttgg ccc23<210>50<211>20<212>DNA<213>人工序列<220><223>人工序列的描述pdhA的引物<400>50cgctggaatccgaacatcga 20<210>51<211>26<212>DNA<213>人工序列<220><223>人工序列的描述pc的引物<400>51ggcgcaacct acgacgttgc aatgcg 26<210>52<211>20<212>DNA<213>人工序列<220><223>人工序列的描述pc的引物<400>52tggccgcctg ggatctcgtg20<210>53<211>20<212>DNA<213>人工序列<220><223>人工序列的描述ppc的引物<400>53ggttcctgga ttggtggaga20<210>54<211>20<212>DNA<213>人工序列<220><223>人工序列的描述ppc的引物<400>54ccgccatcct tgttggaatc20<210>55<211>20<212>DNA<213>人工序列<220><223>人工序列的描述acn的引物<220><221>不确定<222>(3，6，9)<223>n＝次黄苷<400>55gtnggnacng aytcscatac20<210>56<211>20<212>DNA<213>人工序列<220><223>人工序列的描述acn的引物<220><221>不确定<222>(3，9，18)<223>n＝次黄苷<400>56gcnggagana tgtgrtcngt20<210>57<211>20<212>DNA<213>人工序列<220><223>人工序列的描述icd的引物<400>57gacatttcac tcgctggacg20<210>58<211>20<212>DNA<213>人工序列<220><223>人工序列的描述icd的引物<400>58ccgtactctt cagccttctg20<210>59<211>17<212>DNA<213>人工序列<220><223>人工序列的描述lpd的引物<400>59atcatcgcaa ccggttc 17<210>60<211>19<212>DNA<213>人工序列<220><223>人工序列的描述lpd的引物<400>60cgtcaccgat ggcgtaaat 19<210>61<211>20<212>DNA<213>人工序列<220><223>人工序列的描述odhA的引物<400>61acaccgtggt cgcctcaacg20<210>62<211>20<212>DNA<213>人工序列<220><223>人工序列的描述odhA的引物<400>62tgctaacccg tcccacctgg20<210>63<211>20<212>DNA<213>人工序列<220><223>人工序列的描述lpd筛选PCR的引物<400>63tacgaggagc agatcctcaa20<210>64<211>20<212>DNA<213>人工序列<220><223>人工序列的描述lpd筛选PCR的引物<400>64ttgacgccgg tgttctccag20<210>65<211>20<212>DNA<213>人工序列<220><223>人工序列的描述acn LA克隆的引物<400>65ggtgaagcta agtagttagc20<210>66<211>18<212>DNA<213>人工序列<220><223>人工序列的描述acn LA克隆的引物<400>66agctactaaa cctgcacc 18<210>67<211>20<212>DNA<213>人工序列<220><223>人工序列的描述icd LA克隆的引物<400>67ccgtactctt cagccttctg67<210>68<211>18<212>DNA<213>人工序列<220><223>人工序列的描述icd LA克隆的引物<400>68tcgtccttgt tccacatc 18<210>69<211>17<212>DNA<213>人工序列<220><223>人工序列的描述lpd LA克隆的引物<400>69atcatcgcaa ccggttc 17<210>70<211>20<212>DNA<213>人工序列<220><223>人工序列的描述lpd LA克隆的引物<400>70tacgaggagc agatcctcaa20<210>71<211>20<212>DNA<213>人工序列<220><223>人工序列的描述acn LA克隆的引物<400>71gctaactact tagcttcacc20<210>72<211>20<212>DNA<213>人工序列<220><223>人工序列的描述acn LA克隆的引物<400>72gaaccaggaa ctattgaacc20<210>73<211>18<212>DNA<213>人工序列<220><223>人工序列的描述icd LA克隆的引物<400>73tccgatgtca tcatcgac 18<210>74<211>18<212>DNA<213>人工序列<220><223>人工序列的描述icd LA克隆的引物<400>74atgtggaaca aggacgac 18<210>75<211>35<212>DNA<213>人工序列<220><223>人工序列的描述odhA LA克隆的引物<400>75gtacatattg tcgttagaac gcgtaatacg actca 35<210>76<211>35<212>DNA<213>人工序列<220><223>人工序列的描述odhA LA克隆的引物<400>76cgttagaacg cgtaatacga ctcactatag ggaga 35<210>77<211>32<212>DNA<213>人工序列<220><223>人工序列的描述gdh基因扩增引物<400>77gcgcctgcag gtccgagggt gtgcgttcgg ca 32<210>78<211>32<212>DNA<213>人工序列<220><223>人工序列的描述gdh基因扩增引物<400>78gcgcctgcag ccaccagga tgccctcaacc ag 32<210>79<211>1344<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(1)..(1341)<400>79atg act gta gat gag cag gtc tcc aac tac tac gac atg ctg ctg aag 48Met Thr Val Asp Glu Gln Val Ser Asn Tyr Tyr Asp Met Leu Leu Lys 1 5 10 15cgc aac gcc ggg gaa cct gag ttc cac cag gct gtc gcg gag gtt ctc 96Arg Asn Ala Gly Glu Pro Glu Phe His Gln Ala Val Ala Glu Val Leu
20 25 30gaa tct ctg aag atc gtc ctg gag aag gac ccg cac tac gcc gac tac 144Glu Ser Leu Lys Ile Val Leu Glu Lys Asp Pro His Tyr Ala Asp Tyr
35 40 45ggt ctg atc cag cgt ctc tgc gaa ccg gaa cgc cag ctg atc ttc cgt 192Gly Leu Ile Gln Arg Leu Cys Glu Pro Glu Arg Gln Leu Ile Phe Arg
50 55 60gtg ccc tgg gtg gat gac aac ggt cag gtg cac gtc aac cgt ggt ttc 240Val Pro Trp Val Asp Asp Asn Gly Gln Val His Val Asn Arg Gly Phe 65 70 75 80cgt gtc cag ttc aac tcc gca ctc ggc ccg tac aag ggt ggt ctg cgt 288Arg Val Gln Phe Asn Ser Ala Leu Gly Pro Tyr Lys Gly Gly Leu Arg
85 90 95ttc cac ccc tcc gtc aac ctc ggc atc gtc aag ttc ctc ggc ttc gag 336Phe His Pro Ser Val Asn Leu Gly Ile Val Lys Phe Leu Gly Phe Glu
100 105 110cag atc ttc aag aac tcc ctc acc ggt ctg ccg atc ggt ggc ggc aag 384Gln Ile Phe Lys Asn Ser Leu Thr Gly Leu Pro Ile Gly Gly Gly Lys
115 120 125ggt ggt tcc gac ttc gac ccg aag ggc aag tcc gag ctg gag atc atg 432Gly Gly Ser Asp Phe Asp Pro Lys Gly Lys Ser Glu Leu Glu Ile Met
130 135 140cgc ttc tgc cag tcc ttc atg acc gag ctg cac cgc cac atc ggc gag 480Arg Phe Cys Gln Ser Phe Met Thr Glu Leu His Arg His Ile Gly Glu145 150 155 160tac cgg gat gtc ccg gcc ggt gac atc gga gtc ggt ggc cgc gag atc 528Tyr Arg Asp Val Pro Ala Gly Asp Ile Gly Val Gly Gly Arg Glu Ile
165 170 175ggt tac ctc ttc ggc cac tac cgc cgt ctg gcc aac cag cac gag tcc 576Gly Tyr Leu Phe Gly His Tyr Arg Arg Leu Ala Asn Gln His Glu Ser
180 185 190ggt gtg ctc acc ggc aag ggc ctg acc tgg ggt ggt tcc ctg gtc cgc 624Gly Val Leu Thr Gly Lys Gly Leu Thr Trp Gly Gly Ser Leu Val Arg
195 200 205acc gag gcc acc ggc ttc ggc acc gtc tac ttc gtc cag gag atg atc 672Thr Glu Ala Thr Gly Phe Gly Thr Val Tyr Phe Val Gln Glu Met Ile
210 215 220aag gcg gaa ggg gag acc ctc gag ggc aag aag gtc atc gtc tcc ggt720Lys Ala Glu Gly Glu Thr Leu Glu Gly Lys Lys Val Ile Val Ser Gly225 230 235 240tcc ggc aac gtg gcc acc tac gcc atc cag aag gtg cag gaa ctg ggt 768Ser Gly Asn Val Ala Thr Tyr Ala Ile Gln Lys Val Gln Glu Leu Gly
245 250 255gcg gtt gtg gtc ggc ttc tcc gac tcc agc ggc tgg gtc tcc acc ccg 816Ala Val Val Val Gly Phe Ser Asp Ser Ser Gly Trp Val Ser Thr Pro
260 265 270aac ggt gtt gac gtg gcc aag ctg cgt gag atc aag gag gtc cgt cgt 864Asn Gly Val Asp Val Ala Lys Leu Arg Glu Ile Lys Glu Val Arg Arg
275 280 285gca cgc gtg tcc tcc tac gcc gac gag gtg gag ggt gcg gag tac cac 912Ala Arg Val Ser Ser Tyr Ala Asp Glu Val Glu Gly Ala Glu Tyr His
290 295 300acc gac ggc tcc atc tgg gat ctg acc gcc gac atc gcg ctg ccc tgc 960Thr Asp Gly Ser Ile Trp Asp Leu Thr Ala Asp Ile Ala Leu Pro Cys305 310 315 320gcc acc cag aac gaa ctg gac ggc gac aac gcc cgc acc ctc gcg gac 1008Ala Thr Gln Asn Glu Leu Asp Gly Asp Asn Ala Arg Thr Leu Ala Asp
325 330 335aac ggc tgc cgc ttc gtg gcg gag ggc gcc aac atg ccc tcc acc ccc 1056Asn Gly Cys Arg Phe Val Ala Glu Gly Ala Asn Met Pro Ser Thr Pro
340 345 350gag gcc atc gac gtc ttc cgt gag cgt ggt gtt ctc ttc ggg ccg ggc 1104Glu Ala Ile Asp Val Phe Arg Glu Arg Gly Val Leu Phe Gly Pro Gly
355 360 365aag gct gcc aac gcc ggt ggc gtg gcc acc tcc gcc ctg gag atg cag 1152Lys Ala Ala Asn Ala Gly Gly Val Ala Thr Ser Ala Leu Glu Met Gln
370 375 380cag aac gcc tcc cgt gat tcc tgg agc ttc gag tac acc gat gag cgt 1200Gln Asn Ala Ser Arg Asp Ser Trp Ser Phe Glu Tyr Thr Asp Glu Arg385 390 395 400ctc cac cgc atc atg aag aac atc ttc aag tcc tgc gcc gat acc gcc 1248Leu His Arg Ile Met Lys Asn Ile Phe Lys Ser Cys Ala Asp Thr Ala
405 410 415aag gag tac ggc cac gag aag aac tac gtg gtc ggt gcg aac atc gcc 1296Lys Glu Tyr Gly His Glu Lys Asn Tyr Val Val Gly Ala Asn Ile Ala
420 425 430gga ttc aag aag gtc gct gac gcc atg ctc gcc cag ggt gtc atc taa 1344Gly Phe Lys Lys Val Ala Asp Ala Met Leu Ala Gln Gly Val Ile
435 440 445<210>80<211>447<212>PRT<213>Corynebaeterium thermoaminogenes<400>80Met Thr Val Asp Glu Gln Val Ser Asn Tyr Tyr Asp Met Leu Leu Lys 1 5 10 15Arg Asn Ala Gly Glu Pro Glu Phe His Gln Ala Val Ala Glu Val Leu
20 25 30Glu Ser Leu Lys Ile Val Leu Glu Lys Asp Pro His Tyr Ala Asp Tyr
35 40 45Gly Leu Ile Gln Arg Leu Cys Glu Pro Glu Arg Gln Leu Ile Phe Arg
50 55 60Val Pro Trp Val Asp Asp Asn Gly Gln Val His Val Asn Arg Gly Phe 65 70 75 80Arg Val Gln Phe Asn Ser Ala Leu Gly Pro Tyr Lys Gly Gly Leu Arg
85 90 95Phe His Pro Ser Val Asn Leu Gly Ile Val Lys Phe Leu Gly Phe Glu
100 105 110Gln Ile Phe Lys Asn Ser Leu Thr Gly Leu Pro Ile Gly Gly Gly Lys
115 120 125Gly Gly Ser Asp Phe Asp Pro Lys Gly Lys Ser Glu Leu Glu Ile Met
130 135 140Arg Phe Cys Gln Ser Phe Met Thr Glu Leu His Arg His Ile Gly Glu145 150 155 160Tyr Arg Asp Val Pro Ala Gly Asp Ile Gly Val Gly Gly Arg Glu Ile
165 170 175Gly Tyr Leu Phe Gly His Tyr Arg Arg Leu Ala Asn Gln His Glu Ser
180 185 190Gly Val Leu Thr Gly Lys Gly Leu Thr Trp Gly Gly Ser Leu Val Arg
195 200 205Thr Glu Ala Thr Gly Phe Gly Thr Val Tyr Phe Val Gln Glu Met Ile
210 215 220Lys Ala Glu Gly Glu Thr Leu Glu Gly Lys Lys Val Ile Val Ser Gly225 230 235 240Ser Gly Asn Val Ala Thr Tyr Ala Ile Gln Lys Val Gln Glu Leu Gly
245 250 255Ala Val Val Val Gly Phe Ser Asp Ser Ser Gly Trp Val Ser Thr Pro
260 265 270Asn Gly Val Asp Val Ala Lys Leu Arg Glu Ile Lys Glu Val Arg Arg
275 280 285Ala Arg Val Ser Ser Tyr Ala Asp Glu Val Glu Gly Ala Glu Tyr His
290 295 300Thr Asp Gly Ser Ile Trp Asp Leu Thr Ala Asp Ile Ala Leu Pro Cys305 310 315 320Ala Thr Gln Asn Glu Leu Asp Gly Asp Asn Ala Arg Thr Leu Ala Asp
325 330 335Asn Gly Cys Arg Phe Val Ala Glu Gly Ala Asn Met Pro Ser Thr Pro
340 345 350Glu Ala Ile Asp Val Phe Arg Glu Arg Gly Val Leu Phe Gly Pro Gly
355 360 365Lys Ala Ala Asn Ala Gly Gly Val Ala Thr Ser Ala Leu Glu Met Gln
370 375 380Gln Asn Ala Ser Arg Asp Ser Trp Ser Phe Glu Tyr Thr Asp Glu Arg385 390 395 400Leu His Arg Ile Met Lys Asn Ile Phe Lys Ser Cys Ala Asp Thr Ala
405 410 415Lys Glu Tyr Gly His Glu Lys Asn Tyr Val Val Gly Ala Asn Ile Ala
420 425 430Gly Phe Lys Lys Val Ala Asp Ala Met Leu Ala Gln Gly Val Ile
435 440 445<210>81<211>1344<212>DNA<213>乳发酵短杆菌(Brevibacterium lactofermentum)<220><221>CDS<222>(1)..(1341)<400>81atg aca gtt gat gag cag gtc tct aac tat tac gac atg ctt ctg aag 48Met Thr Val Asp Glu Gln Val Ser Asn Tyr Tyr Asp Met Leu Leu Lys 1 5 10 15cgc aat gct ggc gag cct gaa ttt cac cag gca gtg gca gag gtt ttg 96Arg Asn Ala Gly Glu Pro Glu Phe His Gln Ala Val Ala Glu Val Leu
20 25 30gaa tct ttg aag atc gtc ctg gaa aag gac cct cat tac gct gat tac 144Glu Ser Leu Lys Ile Val Leu Glu Lys Asp Pro His Tyr Ala Asp Tyr
35 40 45ggt ctc atc cag cgc ctg tgc gag cct gag cgt cag ctc atc ttc cgt 192Gly Leu Ile Gln Arg Leu Cys Glu Pro Glu Arg Gln Leu Ile Phe Arg
50 55 60gtg cct tgg gtt gat gac cag ggc cag gtc cac gtc aac cgt ggt ttc 240Val Pro Trp Val Asp Asp Gln Gly Gln Val His Val Asn Arg Gly Phe 65 70 75 80cgc gtg cag ttc aac tct gca ctt gga cca tac aag ggc ggc ctg cgc 288Arg Val Gln Phe Asn Ser Ala Leu Gly Pro Tyr Lys Gly Gly Leu Arg
85 90 95ttc cac cca tct gta aac ctg ggc att gtg aag ttc ctg ggc ttt gag 336Phe His Pro Ser Val Asn Leu Gly Ile Val Lys Phe Leu Gly Phe Glu
100 105 110cag atc ttt aaa aac tcc cta acc ggc ctg cca atc ggt ggt ggc aag 384Gln Ile Phe Lys Asn Ser Leu Thr Gly Leu Pro Ile Gly Gly Gly Lys
115 120 125ggt gga tcc gac ttc gac cct aag ggc aag tcc gat ctg gaa atc atg 432Gly Gly Ser Asp Phe Asp Pro Lys Gly Lys Ser Asp Leu Glu Ile Met
130 135 140cgt ttc tgc cag tcc ttc atg acc gag ctg cac cgc cac atc ggt gag 480Arg Phe Cys Gln Ser Phe Met Thr Glu Leu His Arg His Ile Gly Glu145 150 155 160tac cgc gac gtt cct gca ggt gac atc gga gtt ggt ggc cgc gag atc 528Tyr Arg Asp Val Pro Ala Gly Asp Ile Gly Val Gly Gly Arg Glu Ile
165 170 175ggt tac ctg ttt ggc cac tac cgt cgc atg gct aac cag cac gag tcc 576Gly Tyr Leu Phe Gly His Tyr Arg Arg Met Ala Asn Gln His Glu Ser
180 185 190ggc gtt ttg acc ggt aag ggc ctg acc tgg ggt gga tcc ctg gtc cgc 624Gly Val Leu Thr Gly Lys Gly Leu Thr Trp Gly Gly Ser Leu Val Arg
195 200 205acc gag gca act ggc tac ggc tgc gtt tac ttc gtg agt gaa atg atc 672Thr Glu Ala Thr Gly Tyr Gly Cys Val Tyr Phe Val Ser Glu Met Ile
210 215 220aag gct aag ggc gag agc atc agc ggc cag aag atc atc gtt tcc ggt 720Lys Ala Lys Gly Glu Ser Ile Ser Gly Gln Lys Ile Ile Val Ser Gly225 230 235 240tcc ggc aac gta gca acc tac gcg att gaa aag gct cag gaa ctc ggc 768Ser Gly Asn Val Ala Thr Tyr Ala Ile Glu Lys Ala Gln Glu Leu Gly
245 250 255gca acc gtt att ggt ttc tcc gat tcc agc ggt tgg gtt cat acc cct 816Ala Thr Val Ile Gly Phe Ser Asp Ser Ser Gly Trp Val His Thr Pro
260 265 270aac ggc gtt gac gtg gct aag ctc cgc gaa atc aag gaa gtt cgc cgc 864Asn Gly Val Asp Val Ala Lys Leu Arg Glu Ile Lys Glu Val Arg Arg
275 280 285gca cgc gta tcc gtg tac gcc gac gaa att gaa ggc gca acc tac cac 912Ala Arg Val Ser Val Tyr Ala Asp Glu Ile Glu Gly Ala Thr Tyr His
290 295 300acc gac ggt tcc atc tgg gat ctc aag tgc gat atc gct ctt cct tgt 960Thr Asp Gly Ser Ile Trp Asp Leu Lys Cys Asp Ile Ala Leu Pro Cys305 310 315 320gca act cag aac gag ctc aac ggc gag aac gct aag act ctt gca gac 1008Ala Thr Gln Asn Glu Leu Asn Gly Glu Asn Ala Lys Thr Leu Ala Asp
325 330 335aac ggc tgc cgt ttc gtt gct gaa ggc gcg aac atg cct tcc acc cct 1056Asn Gly Cys Arg Phe Val Ala Glu Gly Ala Asn Met Pro Ser Thr Pro
340 345 350gag gct gtt gag gtc ttc cgt gag cgc gac atc cgc ttc gga cca ggc 1104Glu Ala Val Glu Val Phe Arg Glu Arg Asp Ile Arg Phe Gly Pro Gly
355 360 365aag gca gct aac gct ggt ggc gtt gca acc tcc gct ctg gag atg cag 1152Lys Ala Ala Asn Ala Gly Gly Val Ala Thr Ser Ala Leu Glu Met Gln
370 375 380cag aac gct tcg cgc gat tcc tgg agc ttc gag tac acc gac gag cgc 1200Gln Asn Ala Ser Arg Asp Ser Trp Ser Phe Glu Tyr Thr Asp Glu Arg385 390 395 400ctc cag gtg atc atg aag aac atc ttc aag acc tgt gca gag acc gca 1248Leu Gln Val Ile Met Lys Asn Ile Phe Lys Thr Cys Ala Glu Thr Ala
405 410 415gca gag tat gga cac gag aac gat tac gtt gtc ggc gct aac att gct 1296Ala Glu Tyr Gly His Glu Asn Asp Tyr Val Val Gly Ala Asn Ile Ala
420 425 430ggc ttt aag aag gta gct gac gcg atg ctg gca cag ggc gtc atc taa 1344Gly Phe Lys Lys Val Ala Asp Ala Met Leu Ala Gln Gly Val Ile
435 440 445<210>82<211>447<212>PRT<213>乳发酵短杆菌(Brevibacterium lactofermentum)<400>82Met Thr Val Asp Glu Gln Val Ser Asn Tyr Tyr Asp Met Leu Leu Lys 1 5 10 15Arg Asn Ala Gly Glu Pro Glu Phe His Gln Ala Val Ala Glu Val Leu
20 25 30Glu Ser Leu Lys Ile Val Leu Glu Lys Asp Pro His Tyr Ala Asp Tyr
35 40 45Gly Leu Ile Gln Arg Leu Cys Glu Pro Glu Arg Gln Leu Ile Phe Arg
50 55 60Val Pro Trp Val Asp Asp Gln Gly Gln Val His Val Asn Arg Gly Phe 65 70 75 80Arg Val Gln Phe Asn Ser Ala Leu Gly Pro Tyr Lys Gly Gly Leu Arg
85 90 95Phe His Pro Ser Val Asn Leu Gly Ile Val Lys Phe Leu Gly Phe Glu
100 105 110Gln Ile Phe Lys Asn Ser Leu Thr Gly Leu Pro Ile Gly Gly Gly Lys
115 120 125Gly Gly Ser Asp Phe Asp Pro Lys Gly Lys Ser Asp Leu Glu Ile Met
130 135 140Arg Phe Cys Gln Ser Phe Met Thr Glu Leu His Arg His Ile Gly Glu145 150 155 160Tyr Arg Asp Val Pro Ala Gly Asp Ile Gly Val Gly Gly Arg Glu Ile
165 170 175Gly Tyr Leu Phe Gly His Tyr Arg Arg Met Ala Asn Gln His Glu Ser
180 185 190Gly Val Leu Thr Gly Lys Gly Leu Thr Trp Gly Gly Ser Leu Val Arg
195 200 205Thr Glu Ala Thr Gly Tyr Gly Cys Val Tyr Phe Val Ser Glu Met Ile
210 215 220Lys Ala Lys Gly Glu Ser Ile Ser Gly Gln Lys Ile Ile Val Ser Gly225 230 235 240Ser Gly Asn Val Ala Thr Tyr Ala Ile Glu Lys Ala Gln Glu Leu Gly
245 250 255Ala Thr Val Ile Gly Phe Ser Asp Ser Ser Gly Trp Val His Thr Pro
260 265 270Asn Gly Val Asp Val Ala Lys Leu Arg Glu Ile Lys Glu Val Arg Arg
275 280 285Ala Arg Val Ser Val Tyr Ala Asp Glu Ile Glu Gly Ala Thr Tyr His
290 295 300Thr Asp Gly Ser Ile Trp Asp Leu Lys Cys Asp Ile Ala Leu Pro Cys305 310 315 320Ala Thr Gln Asn Glu Leu Asn Gly Glu Asn Ala Lys Thr Leu Ala Asp
325 330 335Asn Gly Cys Arg Phe Val Ala Glu Gly Ala Asn Met Pro Ser Thr Pro
340 345 350Glu Ala Val Glu Val Phe Arg Glu Arg Asp Ile Arg Phe Gly Pro Gly
355 360 365Lys Ala Ala Asn Ala Gly Gly Val Ala Thr Ser Ala Leu Glu Met Gln
370 375 380Gln Asn Ala Ser Arg Asp Ser Trp Ser Phe Glu Tyr Thr Asp Glu Arg385 390 395 400Leu Gln Val Ile Met Lys Asn Ile Phe Lys Thr Cys Ala Glu Thr Ala
405 410 415Ala Glu Tyr Gly His Glu Asn Asp Tyr Val Val Gly Ala Asn Ile Ala
420 425 430Gly Phe Lys Lys Val Ala Asp Ala Met Leu Ala Gln Gly Val Ile
435 440 445<210>83<211>20<212>DNA<213>人工序列<220><223>人工序列的描述gltA基因扩增引物<220><221>misc_feature<222>(9)<223>n＝次黄苷<400>83aagatcacnt acatcgaygg20<210>84<211>20<212>DNA<213>人工序列<220><223>人工序列的描述gltA基因扩增引物<400>84tagaagtcta cgttcgggta20<210>85<211>21<212>DNA<213>人工序列<220><223>人工序列的描述gltA基因扩增引物<400>85gtcgacaata gcctgaatct g 21<210>86<211>21<212>DNA<213>人工序列<220><223>人工序列的描述gltA基因扩增引物<400>86cggtggaacc ggtgctgaca t 21<210>87<211>21<212>DNA<213>人工序列<220><223>人工序列的描述gltA基因扩增引物<400>87gggtgggga attcggtcatg t 21<210>88<211>21<212>DNA<213>人工序列<220><223>人工序列的描述gltA基因扩增引物<400>88tgtcgtagcc gcggtagcgc a 21<210>89<211>1293<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(1)..(1290)<400>89gtg gct tct gat aac aac aag gct gta ctg cac tac cct ggc ggc gaa 48Val Ala Ser Asp Asn Asn Lys Ala Val Leu His Tyr Pro Gly Gly Glu 1 5 10 15ttc gag atg ggc atc aag cag gcc acc gag ggt aac tcc ggt gtc atc 96Phe Glu Met Gly Ile Lys Gln Ala Thr Glu Gly Asn Ser Gly Val Ile
20 25 30ctg ggt aag atg ctg tcg gaa acc ggt ctg gtc acc ttc gac ccc ggt 144Leu Gly Lys Met Leu Ser Glu Thr Gly Leu Val Thr Phe Asp Pro Gly
35 40 45tat gtc agc acc ggt tcc acc gaa tcc aag atc acc tac atc gat ggt 192Tyr Val Ser Thr Gly Ser Thr Glu Ser Lys Ile Thr Tyr Ile Asp Gly
50 55 60gat gca ggc atc ctg cgc tac cgc ggc tac gac att gcg gat ctg gcc 240Asp Ala Gly Ile Leu Arg Tyr Arg Gly Tyr Asp Ile Ala Asp Leu Ala 65 70 75 80gaa aat gcc acc ttc aat gag gtc tcc tac ctc ctg atc aag ggt gag 288Glu Asn Ala Thr Phe Asn Glu Val Ser Tyr Leu Leu Ile Lys Gly Glu
85 90 95ctc ccg acc ccg gaa gag ctc cac aag ttc aac gac gag att cgt cac 336Leu Pro Thr Pro Glu Glu Leu His Lys Phe Asn Asp Glu Ile Arg His
100 105 110cac acc ctg ctg gac gag gac ttc aag tcc cag ttc aat gtc ttc cct 384His Thr Leu Leu Asp Glu Asp Phe Lys Ser Gln Phe Asn Val Phe Pro
115 120 125cgc gat gcc cac ccg atg gcc acc ctg gcc tcc tcg gtt aac atc ctc 432Arg Asp Ala His Pro Met Ala Thr Leu Ala Ser Ser Val Asn Ile Leu
130 135 140tcc acc tac tac cag gat cag ctg gat ccc ctg gat gag gct cag ctg 480Ser Thr Tyr Tyr Gln Asp Gln Leu Asp Pro Leu Asp Glu Ala Gln Leu145 150 155 160gac aag gca acc gtc cgc ctg atg gcg aag gtt ccg atg ctg gct gca 528Asp Lys Ala Thr Val Arg Leu Met Ala Lys Val Pro Met Leu Ala Ala
165 170 175tac gca cac cgt gcc cgc aag ggt gcg ccg tac atg tac ccg gac aac 576Tyr Ala His Arg Ala Arg Lys Gly Ala Pro Tyr Met Tyr Pro Asp Asn
180 185 190tcc ctc aat gcc cgt gag aac ttc ctg cgc atg atg ttc ggt tac ccg 624Ser Leu Asn Ala Arg Glu Asn Phe Leu Arg Met Met Phe Gly Tyr Pro
195 200 205acc gag ccg tac gag gtt gat ccg atc atg gtc aaa gcc ctc gac aag 672Thr Glu Pro Tyr Glu Val Asp Pro Ile Met Val Lys Ala Leu Asp Lys
210 215 220ctg ctc atc ctg cac gca gac cac gag cag aac tgc tcc acc tcc act 720Leu Leu Ile Leu His Ala Asp His Glu Gln Asn Cys Ser Thr Ser Thr225 230 235 240gtc cgc atg atc ggc tcc gcg cag gcg aac atg ttc gtc tcc atc gcc 768Val Arg Met Ile Gly Ser Ala Gln Ala Asn Met Phe Val Ser Ile Ala
245 250 255ggc ggc atc aac gca ctc tcc ggc ccg ctg cac ggt ggc gcc aac cag 816Gly Gly Ile Asn Ala Leu Ser Gly Pro Leu His Gly Gly Ala Asn Gln
260 265 270gct gtc ctc gag atg ctc gag gag atc gca gcc aac ggc ggc gac gca 864Ala Val Leu Glu Met Leu Glu Glu Ile Ala Ala Asn Gly Gly Asp Ala
275 280 285acc gac ttc atg aac cgc gtg aag aac aag gag aag ggt gtc cgc ctc 912Thr Asp Phe Met Asn Arg Val Lys Asn Lys Glu Lys Gly Val Arg Leu
290 295 300atg ggc ttc gga cac cgc gtc tac aag aac tac gat ccg cgt gca gcc 960Met Gly Phe Gly His Arg Val Tyr Lys Asn Tyr Asp Pro Arg Ala Ala305 310 315 320atc gtc aag gac acc gcc cac gag atc ctc gag cac ctc ggt ggc gac 1008Ile Val Lys Asp Thr Ala His Glu Ile Leu Glu His Leu Gly Gly Asp
325 330 335cca ctg ctg gat ctg gct ctc aag ctg gaa gaa atc gca ctc aac gac 1056Pro Leu Leu Asp Leu Ala Leu Lys Leu Glu Glu Ile Ala Leu Asn Asp
340 345 350gat tac ttc atc tcc cgc aag ctg tac ccg aac gtg gac ttc tac acc 1104Asp Tyr Phe Ile Ser Arg Lys Leu Tyr Pro Asn Val Asp Phe Tyr Thr
355 360 365ggc ctg atc tac cgc gcc atg ggc ttc ccg acg gac ttc ttc acc gtc 1152Gly Leu Ile Tyr Arg Ala Met Gly Phe Pro Thr Asp Phe Phe Thr Val
370 375 380ctg ttc gcc atc ggc cgc ctc ccg ggc tgg atc gcc cac tac cgc gag 1200Leu Phe Ala Ile Gly Arg Leu Pro Gly Trp Ile Ala His Tyr Arg Glu385 390 395 400cag ctc gcc gat ccg ggc gcc aag atc aac cgt cct cgc cag atc tac 1248Gln Leu Ala Asp Pro Gly Ala Lys Ile Asn Arg Pro Arg Gln Ile Tyr
405 410 415acc ggt gag acc gca cgc aag atc atc ccc cgc gaa gag cgc tag 1293Thr Gly Glu Thr Ala Arg Lys Ile Ile Pro Arg Glu Glu Arg
420 425 430<210>90<211>430<212>PRT<213>Corynebacterium thermoaminogenes<400>90Val Ala Ser Asp Asn Asn Lys Ala Val Leu His Tyr Pro Gly Gly Glu 1 5 10 15Phe Glu Met Gly Ile Lys Gln Ala Thr Glu Gly Asn Ser Gly Val Ile
20 25 30Leu Gly Lys Met Leu Ser Glu Thr Gly Leu Val Thr Phe Asp Pro Gly
35 40 45Tyr Val Ser Thr Gly Ser Thr Glu Ser Lys Ile Thr Tyr Ile Asp Gly
50 55 60Asp Ala Gly Ile Leu Arg Tyr Arg Gly Tyr Asp Ile Ala Asp Leu Ala 65 70 75 80Glu Asn Ala Thr Phe Asn Glu Val Ser Tyr Leu Leu Ile Lys Gly Glu
85 90 95Leu Pro Thr Pro Glu Glu Leu His Lys Phe Asn Asp Glu Ile Arg His
100 105 110His Thr Leu Leu Asp Glu Asp Phe Lys Ser Gln Phe Asn Val Phe Pro
115 120 125Arg Asp Ala His Pro Met Ala Thr Leu Ala Ser Ser Val Asn Ile Leu
130 135 140Ser Thr Tyr Tyr Gln Asp Gln Leu Asp Pro Leu Asp Glu Ala Gln Leu145 150 155 160Asp Lys Ala Thr Val Arg Leu Met Ala Lys Val Pro Met Leu Ala Ala
165 170 175Tyr Ala His Arg Ala Arg Lys Gly Ala Pro Tyr Met Tyr Pro Asp Asn
180 185 190Ser Leu Asn Ala Arg Glu Asn Phe Leu Arg Met Met Phe Gly Tyr Pro
195 200 205Thr Glu Pro Tyr Glu Val Asp Pro Ile Met Val Lys Ala Leu Asp Lys
210 215 220Leu Leu Ile Leu His Ala Asp His Glu Gln Asn Cys Ser Thr Ser Thr225 230 235 240Val Arg Met Ile Gly Ser Ala Gln Ala Asn Met Phe Val Ser Ile Ala
245 250 255Gly Gly Ile Asn Ala Leu Ser Gly Pro Leu His Gly Gly Ala Asn Gln
260 265 270Ala Val Leu Glu Met Leu Glu Glu Ile Ala Ala Asn Gly Gly Asp Ala
275 280 285Thr Asp Phe Met Asn Arg Val Lys Asn Lys Glu Lys Gly Val Arg Leu
290 295 300Met Gly Phe Gly His Arg Val Tyr Lys Asn Tyr Asp Pro Arg Ala Ala305 310 315 320Ile Val Lys Asp Thr Ala His Glu Ile Leu Glu His Leu Gly Gly Asp
325 330 335Pro Leu Leu Asp Leu Ala Leu Lys Leu Glu Glu Ile Ala Leu Asn Asp
340 345 350Asp Tyr Phe Ile Ser Arg Lys Leu Tyr Pro Asn Val Asp Phe Tyr Thr
355 360 365Gly Leu Ile Tyr Arg Ala Met Gly Phe Pro Thr Asp Phe Phe Thr Val
370 375 380Leu Phe Ala Ile Gly Arg Leu Pro Gly Trp Ile Ala His Tyr Arg Glu385 390 395 400Gln Leu Ala Asp Pro Gly Ala Lys Ile Asn Arg Pro Arg Gln Ile Tyr
405 410 415Thr Gly Glu Thr Ala Arg Lys Ile Ile Pro Arg Glu Glu Arg
420 425 430<210>91<211>1314<212>DNA<213>乳发酵短杆菌(Brevibacterium lactofermentum)<220><221>CDS<222>(1)..(1311)<400>91atg ttt gaa agg gat atc gtg gct act gat aac aac aag gct gtc ctg 48Met Phe Glu Arg Asp Ile Val Ala Thr Asp Asn Asn Lys Ala Val Leu 1 5 10 15cac tac ccc ggt ggc gag ttc gaa atg gac atc atc gag gct tct gag 96His Tyr Pro Gly Gly Glu Phe Glu Met Asp Ile Ile Glu Ala Ser Glu
20 25 30ggt aac aac ggt gtt gtc ctg ggc aag atg ctg tct gag act gga ctg 144Gly Asn Asn Gly Val Val Leu Gly Lys Met Leu Ser Glu Thr Gly Leu
35 40 45atc act ttt gac cca ggt tat gtg agc act ggc tcc acc gag tcg aag 192Ile Thr Phe Asp Pro Gly Tyr Val Ser Thr Gly Ser Thr Glu Ser Lys
50 55 60atc acc tac atc gat ggc gat gcg gga atc ctg cgt tac cgc ggc tat 240Ile Thr Tyr Ile Asp Gly Asp Ala Gly Ile Leu Arg Tyr Arg Gly Tyr 65 70 75 80gac atc gct gat ctg gct gag aat gcc acc ttc aac gag gtt tct tac 288Asp Ile Ala Asp Leu Ala Glu Asn Ala Thr Phe Asn Glu Val Ser Tyr
85 90 95cta ctt atc aac ggt gaa cta cca acc cca gat gag ctt cac aag ttt 336Leu Leu Ile Asn Gly Glu Leu Pro Thr Pro Asp Glu Leu His Lys Phe
100 105 110aac gac gag att cgc cac cac acc ctt ctg gac gag gac ttc aag tcc 384Asn Asp Glu Ile Arg His His Thr Leu Leu Asp Glu Asp Phe Lys Ser
115 120 125cag ttc aac gtg ttc cca cgc gac gct cac cca atg gca acc ttg gct 432Gln Phe Asn Val Phe Pro Arg Asp Ala His Pro Met Ala Thr Leu Ala
130 135 140tcc tcg gtt aac att ttg tct acc tac tac cag gat cag ctg aac cca 480Ser Ser Val Asn Ile Leu Ser Thr Tyr Tyr Gln Asp Gln Leu Asn Pro145 150 155 160ctc gat gag gca cag ctt gat aag gca acc gtt cgc ctc atg gca aag 528Leu Asp Glu Ala Gln Leu Asp Lys Ala Thr Val Arg Leu Met Ala Lys
165 170 175gtt cca atg ctg gct gcg tac gca cac cgc gca cgc aag ggt gct cct 576Val Pro Met Leu Ala Ala Tyr Ala His Arg Ala Arg Lys Gly Ala Pro
180 185 190tac atg tac cca gac aac tcc ctc aac gcg cgt gag aac ttc ctg cgc 624Tyr Met Tyr Pro Asp Asn Ser Leu Asn Ala Arg Glu Asn Phe Leu Arg
195 200 205atg atg ttc ggt tac cca acc gag cca tac gag atc gac cca atc atg 672Met Met Phe Gly Tyr Pro Thr Glu Pro Tyr Glu Ile Asp Pro Ile Met
210 215 220gtc aag gct ctg gac aag ctg ctc atc ctg cac gct gac cac gag cag 720Val Lys Ala Leu Asp Lys Leu Leu Ile Leu His Ala Asp His Glu Gln225 230 235 240aac tgc tcc acc tcc acc gtt cgt atg atc ggt tcc gca cag gcc aac 768Asn Cys Ser Thr Ser Thr Val Arg Met Ile Gly Ser Ala Gln Ala Asn
245 250 255atg ttt gtc tcc atc gct ggt ggc atc aac gct ctg tcc ggc cca ctg 816Met Phe Val Ser Ile Ala Gly Gly Ile Asn Ala Leu Ser Gly Pro Leu
260 265 270cac ggt ggc gca aac cag gct gtt ctg gag atg ctc gaa gac atc aag 864His Gly Gly Ala Asn Gln Ala Val Leu Glu Met Leu Glu Asp Ile Lys
275 280 285aac aac cac ggt ggc gac gca acc gcg ttc atg aac aag gtc aag aac 912Asn Asn His Gly Gly Asp Ala Thr Ala Phe Met Asn Lys Val Lys Asn
290 295 300aag gaa gac ggc gtc cgc ctc atg ggc ttc gga cac cgc gtt tac aag 960Lys Glu Asp Gly Val Arg Leu Met Gly Phe Gly His Arg Val Tyr Lys305 310 315 320aac tac gat cca cgt gca gca atc gtc aag gag acc gca cac gag atc 1008Asn Tyr Asp Pro Arg Ala Ala Ile Val Lys Glu Thr Ala His Glu Ile
325 330 335ctc gag cac ctc ggt ggc gac gat ctt ctg gat ctg gca atc aag ctg 1056Leu Glu His Leu Gly Gly Asp Asp Leu Leu Asp Leu Ala Ile Lys Leu
340 345 350gaa gaa att gca ctg gct gat gat tac ttc atc tcc cgc aag ctc tac 1104Glu Glu Ile Ala Leu Ala Asp Asp Tyr Phe Ile Ser Arg Lys Leu Tyr
355 360 365ccg aac gta gac ttc tac acc ggc ctg atc tac cgc gca atg ggc ttc 1152Pro Asn Val Asp Phe Tyr Thr Gly Leu Ile Tyr Arg Ala Met Gly Phe
370 375 380cca act gac ttc ttc acc gta ttg ttc gca atc ggt cgt ctg cca gga 1200Pro Thr Asp Phe Phe Thr Val Leu Phe Ala Ile Gly Arg Leu Pro Gly385 390 395 400tgg atc gct cac tac cgc gag cag ctc ggt gca gca ggc aac aag atc 1248Trp Ile Ala His Tyr Arg Glu Gln Leu Gly Ala Ala Gly Asn Lys Ile
405 410 415aac cgc cca cgc cag gtc tac acc ggc aag gaa tcc cgc aag ttg gtt 1296Asn Arg Pro Arg Gln Val Tyr Thr Gly Lys Glu Ser Arg Lys Leu Val
420 425 430cct cgc gag gag cgc taa 1314Pro Arg Glu Glu Arg
435<210>92<211>437<212>PRT<213>乳发酵短杆菌(Brevibacterium lactofermentum)<400>92Met Phe Glu Arg Asp Ile Val Ala Thr Asp Asn Asn Lys Ala Val Leu 1 5 10 15His Tyr Pro Gly Gly Glu Phe Glu Met Asp Ile Ile Glu Ala Ser Glu
20 25 30Gly Asn Asn Gly Val Val Leu Gly Lys Met Leu Ser Glu Thr Gly Leu
35 40 45Ile Thr Phe Asp Pro Gly Tyr Val Ser Thr Gly Ser Thr Glu Ser Lys
50 55 60Ile Thr Tyr Ile Asp Gly Asp Ala Gly Ile Leu Arg Tyr Arg Gly Tyr 65 70 75 80Asp Ile Ala Asp Leu Ala Glu Asn Ala Thr Phe Asn Glu Val Ser Tyr
85 90 95Leu Leu Ile Asn Gly Glu Leu Pro Thr Pro Asp Glu Leu His Lys Phe
100 105 110Asn Asp Glu Ile Arg His His Thr Leu Leu Asp Glu Asp Phe Lys Ser
115 120 125Gln Phe Asn Val Phe Pro Arg Asp Ala His Pro Met Ala Thr Leu Ala
130 135 140Ser Ser Val Asn Ile Leu Ser Thr Tyr Tyr Gln Asp Gln Leu Asn Pro145 150 155 160Leu Asp Glu Ala Gln Leu Asp Lys Ala Thr Val Arg Leu Met Ala Lys
165 170 175Val Pro Met Leu Ala Ala Tyr Ala His Arg Ala Arg Lys Gly Ala Pro
180 185 190Tyr Met Tyr Pro Asp Asn Ser Leu Asn Ala Arg Glu Asn Phe Leu Arg
195 200 205Met Met Phe Gly Tyr Pro Thr Glu Pro Tyr Glu Ile Asp Pro Ile Met
210 215 220Val Lys Ala Leu Asp Lys Leu Leu Ile Leu His Ala Asp His Glu Gln225 230 235 240Asn Cys Ser Thr Ser Thr Val Arg Met Ile Gly Ser Ala Gln Ala Asn
245 250 255Met Phe Val Ser Ile Ala Gly Gly Ile Asn Ala Leu Ser Gly Pro Leu
260 265 270His Gly Gly Ala Asn Gln Ala Val Leu Glu Met Leu Glu Asp Ile Lys
275 280 285Asn Asn His Gly Gly Asp Ala Thr Ala Phe Met Asn Lys Val Lys Asn
290 295 300Lys Glu Asp Gly Val Arg Leu Met Gly Phe Gly His Arg Val Tyr Lys305 310 315 320Asn Tyr Asp Pro Arg Ala Ala Ile Val Lys Glu Thr Ala His Glu Ile
325 330 335Leu Glu His Leu Gly Gly Asp Asp Leu Leu Asp Leu Ala Ile Lys Leu
340 345 350Glu Glu Ile Ala Leu Ala Asp Asp Tyr Phe Ile Ser Arg Lys Leu Tyr
355 360 365Pro Asn Val Asp Phe Tyr Thr Gly Leu Ile Tyr Arg Ala Met Gly Phe
370 375 380Pro Thr Asp Phe Phe Thr Val Leu Phe Ala Ile Gly Arg Leu Pro Gly385 390 395 400Trp Ile Ala His Tyr Arg Glu Gln Leu Gly Ala Ala Gly Asn Lys Ile
405 410 415Asn Arg Pro Arg Gln Val Tyr Thr Gly Lys Glu Ser Arg Lys Leu Val
420 425 430Pro Arg Glu Glu Arg
435<210>93<211>1656<212>DNA<213>Corynebacterium thermoaminogenes<220><221>CDS<222>(309)..(1595)<400>93acgcccgatt cttcaacact atcgaagagg tcccaaccca cgcgttgacc cagggcttgg 60gtactttgtc ccgcgcgcaa aatatcgtgt tggtggcaac tggccaagga aaagcagaca 120gccatccgcg gaactgtgga aggtccagtg actgcttctt gcccaggttc cattctgcaa 180atgcacaaca acgccaccat catcgttgat gaagcagcag catccaagct gaaaaatgct 240gaccattacc gtctcatgga gcaattaaag ctgcgctaga aacaaaaagg aaagtactgt 300gtggggct atg cac aca gaa ctt tcc agt ttg cgc cct gcg tac cat gtg 350
Met His Thr Glu Leu Ser Ser Leu Arg Pro Ala Tyr His Val
1 5 10act cct ccg cag ggc aga ctc aat gat ccc aat gga atg tac gtc gat 398Thr Pro Pro Gln Gly Arg Leu Asn Asp Pro Asn Gly Met Tyr Val Asp 15 20 25 30gga gat acc ctc cac gtc tac tac cag cac gat cca ggt ttc ccc ttc 446Gly Asp Thr Leu His Val Tyr Tyr Gln His Asp Pro Gly Phe Pro Phe
35 40 45gca cca aag cgc acc ggt tgg gct cac acc acc acg ccg ttg acc gga 494Ala Pro Lys Arg Thr Gly Trp Ala His Thr Thr Thr Pro Leu Thr Gly
50 55 60ccg cag cga ttg cag tgg acg cac ctg ccc gat gct ctt tac ccg gat 542Pro Gln Arg Leu Gln Trp Thr His Leu Pro Asp Ala Leu Tyr Pro Asp
65 70 75gta tcc tat gac ctg gat gga tgc tat tcc ggc gga gcc gta ttt tct 590Val Ser Tyr Asp Leu Asp Gly Cys Tyr Ser Gly Gly Ala Val Phe Ser
80 85 90gac ggc acg ctt aaa ctt ttc tac acc ggc aac cga aaa att gac ggc 638Asp Gly Thr Leu Lys Leu Phe Tyr Thr Gly Asn Arg Lys Ile Asp Gly 95 100 105 110aag cgc cgc gcc acc caa aac ctc gtc gaa gtc gag gac cca act ggg 686Lys Arg Arg Ala Thr Gln Asn Leu Val Glu Val Glu Asp Pro Thr Gly
115 120 125ctg atg ggc ggc att cat cgc cgc tcg cct aaa aat ccg ctt atc gac 734Leu Met Gly Gly Ile His Arg Arg Ser Pro Lys Asn Pro Leu Ile Asp
130 135 140gga ccc gcc agc ggt ttt acg ccc cac tac cgc gat ccc atg atc agc 782Gly Pro Ala Ser Gly Phe Thr Pro His Tyr Arg Asp Pro Met Ile Ser
145 150 155cct gat ggg gat ggt tgg aag atg gtt ctt ggg gct cag cgc gaa aac 830Pro Asp Gly Asp Gly Trp Lys Met Val Leu Gly Ala Gln Arg Glu Asn
160 165 170ctc acc ggt gca gcg gtt cta tac cgc tcg gca gat ctt gaa aac tgg 878Leu Thr Gly Ala Ala Val Leu Tyr Arg Ser Ala Asp Leu Glu Asn Trp175 180 185 190gaa ttc tcc ggt gaa atc acc ttt gac ctc agc gac gca caa cct ggt 926Glu Phe Ser Gly Glu Ile Thr Phe Asp Leu Ser Asp Ala Gln Pro Gly
195 200 205tct gcc cct gat ctc gtt cct ggc ggc tac atg tgg gaa tgc ccc aac 974Ser Ala Pro Asp Leu Val Pro Gly Gly Tyr Met Trp Glu Cys Pro Asn
210 215 220ctt ttt acg ctt cgc gat gaa aaa acc ggc gaa gac ctc gat gtg ctg 1022Leu Phe Thr Leu Arg Asp Glu Lys Thr Gly Glu Asp Leu Asp Val Leu
225 230 235att ttc tgt cca caa gga ttg gac cgt atc gat gat gag gtt act cac 1070Ile Phe Cys Pro Gln Gly Leu Asp Arg Ile Asp Asp Glu Val Thr His
240 245 250tac gca agc tct gac cag tgc gga tat gtc gtc ggc aag ctt gaa gaa 1118Tyr Ala Ser Ser Asp Gln Cys Gly Tyr Val Val Gly Lys Leu Glu Glu255 260 265 270acg acc ttc cgt gtc ctg cga gga ttc agc gag ctg gat ttc ggt cat 1166Thr Thr Phe Arg Val Leu Arg Gly Phe Ser Glu Leu Asp Phe Gly His
275 280 285gaa ttc tac gcg ccg cag gtt gca gtc aac ggt tcc gat gcc tgg ctt 1214Glu Phe Tyr Ala Pro Gln Val Ala Val Asn Gly Ser Asp Ala Trp Leu
290 295 300gtg ggc tgg atg gga ttg cct gca cag gat gat cac cca aca gtt gcg 1262Val Gly Trp Met Gly Leu Pro Ala Gln Asp Asp His Pro Thr Val Ala
305 310 315cag gaa gga tgg gtg cac tgc ctg acc gtt cct cgc agg ctt cat ttg 1310Gln Glu Gly Trp Val His Cys Leu Thr Val Pro Arg Arg Leu His Leu
320 325 330cgt aac cat gcg atc tat caa gag ctt ctt ctc cca gaa ggg gag tcg 1358Arg Asn His Ala Ile Tyr Gln Glu Leu Leu Leu Pro Glu Gly Glu Ser335 340 345 350ggg gta act aga tct gta tta ggt tct gaa cct gtc cga gta gac atc 1406Gly Val Thr Arg Ser Val Leu Gly Ser Glu Pro Val Arg Val Asp Ile
355 360 365cga gac aat gtt tcc ctc gag tgg gat ggt gtc cgg ttg tct gtg gat 1454Arg Asp Asn Val Ser Leu Glu Trp Asp Gly Val Arg Leu Ser Val Asp
370 375 380cgc gat ggc gat cgt cgt gta gct gaa gta aaa cct ggc gaa tta gtg 1502Arg Asp Gly Asp Arg Arg Val Ala Glu Val Lys Pro Gly Glu Leu Val
385 390 395atc gcg gac gat aat aca gcg att gag ata aca gca ggt cat ggc cag 1550Ile Ala Asp Asp Asn Thr Ala Ile Glu Ile Thr Ala Gly His Gly Gln
400 405 410gtt tcc ttc gct ttc cgc acc ttc aaa ggt gac act att gag aga1595Val Ser Phe Ala Phe Arg Thr Phe Lys Gly Asp Thr Ile Glu Arg415 420 425taagtcataa aaaagggcct tctgtggcgg attgtacaaa tacttcgcaa aatcccttga 1655t 1656<210>94<211>429<212>PRT<213>Corynebacterium thermoaminogenes<400>94Met His Thr Glu Leu Ser Ser Leu Arg Pro Ala Tyr His Val Thr Pro 1 5 10 15Pro Gln Gly Arg Leu Asn Asp Pro Asn Gly Met Tyr Val Asp Gly Asp
20 25 30Thr Leu His Val Tyr Tyr Gln His Asp Pro Gly Phe Pro Phe Ala Pro
35 40 45Lys Arg Thr Gly Trp Ala His Thr Thr Thr Pro Leu Thr Gly Pro Gln
50 55 60Arg Leu Gln Trp Thr His Leu Pro Asp Ala Leu Tyr Pro Asp Val Ser 65 70 75 80Tyr Asp Leu Asp Gly Cys Tyr Ser Gly Gly Ala Val Phe Ser Asp Gly
85 90 95Thr Leu Lys Leu Phe Tyr Thr Gly Asn Arg Lys Ile Asp Gly Lys Arg
100 105 110Arg Ala Thr Gln Asn Leu Val Glu Val Glu Asp Pro Thr Gly Leu Met
115120 125Gly Gly Ile His Arg Arg Ser Pro Lys Asn Pro Leu Ile Asp Gly Pro
130 135 140Ala Ser Gly Phe Thr Pro His Tyr Arg Asp Pro Met Ile Ser Pro Asp145 150 155 160Gly Asp Gly Trp Lys Met Val Leu Gly Ala Gln Arg Glu Asn Leu Thr
165 170 175Gly Ala Ala Val Leu Tyr Arg Ser Ala Asp Leu Glu Asn Trp Glu Phe
180 185 190Ser Gly Glu Ile Thr Phe Asp Leu Ser Asp Ala Gln Pro Gly Ser Ala
195 200 205Pro Asp Leu Val Pro Gly Gly Tyr Met Trp Glu Cys Pro Asn Leu Phe
210 215 220Thr Leu Arg Asp Glu Lys Thr Gly Glu Asp Leu Asp Val Leu Ile Phe225 230 235 240Cys Pro Gln Gly Leu Asp Arg Ile Asp Asp Glu Val Thr His Tyr Ala
245 250 255Ser Ser Asp Gln Cys Gly Tyr Val Val Gly Lys Leu Glu Glu Thr Thr
260 265 270Phe Arg Val Leu Arg Gly Phe Ser Glu Leu Asp Phe Gly His Glu Phe
275 280 285Tyr Ala Pro Gln Val Ala Val Asn Gly Ser Asp Ala Trp Leu Val Gly
290 295 300Trp Met Gly Leu Pro Ala Gln Asp Asp His Pro Thr Val Ala Gln Glu305 310 315 320Gly Trp Val His Cys Leu Thr Val Pro Arg Arg Leu His Leu Arg Asn
325 330 335His Ala Ile Tyr Gln Glu Leu Leu Leu Pro Glu Gly Glu Ser Gly Val
340 345 350Thr Arg Ser Val Leu Gly Ser Glu Pro Val Arg Val Asp Ile Arg Asp
355 360 365Asn Val Ser Leu Glu Trp Asp Gly Val Arg Leu Ser Val Asp Arg Asp
370 375 380Gly Asp Arg Arg Val Ala Glu Val Lys Pro Gly Glu Leu Val Ile Ala385 390 395 400Asp Asp Asn Thr Ala Ile Glu Ile Thr Ala Gly His Gly Gln Val Ser
405 410 415Phe Ala Phe Arg Thr Phe Lys Gly Asp Thr Ile Glu Arg
420 425<210>95<211>35<212>DNA<213>人工序列<220><223>人工序列的描述scrB基因扩增引物<400>95gtacatattg tcgttagaac gcgtaatacg actca 35<210>96<211>35<212>DNA<213>人工序列<220><223>人工序列的描述scrB基因扩增引物<400>96cgttagaacg cgtaatacga ctcactatag ggaga 35<210>97<211>30<212>DNA<213>人工序列<220><223>人工序列的描述scrB LA克隆引物<400>97gtaaagagcg tcgggcaggt gcgtccactg 30<210>98<211>30<212>DNA<213>人工序列<220><223>人工序列的描述scrB LA克隆引物<400>98ggtgtgagcc cagccggtgc gctttggtgc 30<210>99<211>30<212>DNA<213>人工序列<220><223>人工序列的描述scrB LA克隆引物<400>99atcagccctg atggtgatgg ttggaaaatg 30<210>100<211>30<212>DNA<213>人工序列<220><223>人工序列的描述scrB基因LA克隆引物<400>100ggtgcagcgg ttctataccg ctcgacagat 30<210>101<211>32<212>DNA<213>人工序列<220><223>人工序列的描述scrB基因扩增引物<400>101ggcccgggac gcccgattct tcaacactat cg 32<210>102<211>32<212>DNA<213>人工序列<220><223>人工序列的描述scrB基因扩增引物<400>102ggcccgggga tcaagggatt ttgcgaagta tt 32<210>103<211>30<212>DNA<213>人工序列<220><223>人工序列的描述icd基因扩增引物<400>103gaagatctct atgaccagcg catcaagctg 30<210>104<211>30<212>DNA<213>人工序列<220><223>人工序列的描述icd基因扩增引物<400>104gaagatctgg tcatcccaga acctgatcac 30<210>105<211>32<212>DNA<213>人工序列<220><223>人工序列的描述gdh基因扩增引物<400>105gcgcctgcag gtccgagggt gtgcgttcgg ca 32<210>106<211>32<212>DNA<213>人工序列<220><223>人工序列的描述gdh基因扩增引物<400>106gcgcctgcag gcaccaggat gccctcaacc ag 32<210>107<211>30<212>DNA<213>人工序列<220><223>人工序列的描述gltA基因扩增引物<400>107ggggtaccga tcactataac cccacagcac 30<210>108<211>30<212>DNA<213>人工序列<220><223>人工序列的描述gltA基因扩增引物<400>108ggggtaccct ggctgatctg aactaggcgc 30
权利要求
1.一种蛋白质，该蛋白质具有SEQ ID NO2的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO2的氨基酸序列，该蛋白质具有异柠檬酸裂合酶活性并且在经50℃热处理5分钟后表现出30％或者更高的残余活性。
2.一种蛋白质，该蛋白质具有SEQ ID NO4的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO4的氨基酸序列，该蛋白质涉及到脂酰辅酶A羧化酶活性并且来自于Corynebacterium thermoaminogenes。
3.一种蛋白质，该蛋白质具有SEQ ID NO6的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO6的氨基酸序列，该蛋白质具有DtsR活性并且来自于Corynebacterium thermoaminogenes。
4.一种蛋白质，该蛋白质具有SEQ ID NO8的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO8的氨基酸序列，该蛋白质具有DtsR活性并且来自于Corynebacterium thermoaminogenes。
5.一种蛋白质，该蛋白质具有SEQ ID NO10的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO10的氨基酸序列，该蛋白质在60℃下所表现出的磷酸果糖激酶活性与30℃下的活性相同或者比后者更高。
6.一种蛋白质，该蛋白质具有SEQ ID NO94的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO94的氨基酸序列，该蛋白质具有赋予蔗糖同化能力的活性并且来自于Corynebacterium thermoaminogenes。
7.一种蛋白质，该蛋白质具有SEQ ID NO17-20的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO17-20的氨基酸序列，该蛋白质具有参与谷氨酸摄入的功能并且来自于Corynebacterium thermoaminogenes。
8.一种蛋白质，该蛋白质具有SEQ ID NO22的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO22的氨基酸序列，该蛋白质具有丙酮酸脱氢酶活性并且来自于Corynebacterium thermoaminogenes。
9.一种蛋白质，该蛋白质具有SEQ ID NO24的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO24的氨基酸序列，该蛋白质具有丙酮酸羧化酶活性并且来自于Corynebacterium thermoaminogenes。
10.一种蛋白质，该蛋白质具有SEQ ID NO26的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO26的氨基酸序列，该蛋白质具有磷酸烯醇式丙酮酸羧化酶活性并且在经45℃热处理5分钟后表现出50％或者更高的残余活性。
11.一种蛋白质，该蛋白质具有SEQ ID NO28的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO28的氨基酸序列，该蛋白质具有顺乌头酸酶活性并且在经50℃热处理3分钟后表现出30％或者更高的残余活性。
12.一种蛋白质，该蛋白质具有SEQ ID NO30的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO30的氨基酸序列，该蛋白质具有异柠檬酸脱氢酶活性并且在经45℃热处理10分钟后表现出50％或者更高的残余活性。
13.一种蛋白质，该蛋白质具有SEQ ID NO32的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO32的氨基酸序列，该蛋白质具有二氢硫辛酰胺脱氢酶的活性并且来自于Corynebacterium thermoaminogenes。
14.一种蛋白质，该蛋白质具有SEQ ID NO34的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO34的氨基酸序列，该蛋白质具有2-酮戊二酸脱氢酶活性并且在经50℃热处理10分钟后表现出30％或者更高的残余活性。
15.一种蛋白质，该蛋白质具有SEQ ID NO80的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO80的氨基酸序列，该蛋白质在42℃下所表现出的谷氨酸脱氢酶活性与37℃下的活性相同或者比后者更高。
16.一种蛋白质，该蛋白质具有SEQ ID NO90的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO90的氨基酸序列，该蛋白质在37℃下所表现出的柠檬酸合成酶活性与23℃下的活性相同或者比后者更高。
17.一种DNA，该DNA编码的蛋白质具有SEQ ID NO2的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO2的氨基酸序列，并且该蛋白质具有异柠檬酸裂合酶活性。
18.权利要求17的DNA，该DNA为以下(a1)或者(b1)中所定义的DNA
(a1)一种DNA，该DNA具有序列表中SEQ ID NO1的核苷酸序列，
(b1)一种DNA，该DNA同序列表中的SEQ ID NO1的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有异柠檬酸裂合酶活性的蛋白质。
19.一种DNA，该DNA编码的蛋白质具有SEQ ID NO4的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO4的氨基酸序列，并且该蛋白质涉及到脂酰辅酶A羧化酶活性。
20.权利要求19的DNA，该DNA为以下(a2)或者(b2)中所定义的DNA
(a2)一种DNA，该DNA具有序列表中SEQ ID NO3的核苷酸序列，
(b2)一种DNA，该DNA同序列表中的SEQ ID NO3的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种涉及到脂酰辅酶A羧化酶活性的蛋白质。
21.一种DNA，该DNA编码的蛋白质具有SEQ ID NO6的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO6的氨基酸序列，并且该蛋白质具有DtsR活性。
22.权利要求21的DNA，该DNA为以下(a3)或者(b3)中所定义的DNA
(a3)一种DNA，该DNA具有序列表中SEQ ID NO5的核苷酸序列，
(b3)一种DNA，该DNA同序列表中的SEQ ID NO5的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有DtsR活性的蛋白质。
23.一种DNA，该DNA编码的蛋白质具有SEQ ID NO8的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO8的氨基酸序列，并且该蛋白质具有DtsR活性。
24.权利要求23的DNA，该DNA为以下(a4)或者(b4)中所定义的DNA
(a4)一种DNA，该DNA具有序列表中SEQ ID NO7的核苷酸序列，
(b4)一种DNA，该DNA同序列表中的SEQ ID NO7的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有DtsR活性的蛋白质。
25.一种DNA，该DNA编码的蛋白质具有SEQ ID NO10的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO10的氨基酸序列，并且该蛋白质具有磷酸果糖激酶活性。
26.权利要求25的DNA，该DNA为以下(a5)或者(b5)中所定义的DNA
(a5)一种DNA，该DNA具有序列表中SEQ ID NO9的核苷酸序列，
(b5)一种DNA，该DNA同序列表中的SEQ ID NO9的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有磷酸果糖激酶活性的蛋白质。
27.一种DNA，该DNA编码的蛋白质具有SEQ ID NO93的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO93的氨基酸序列，并且该蛋白质具有蔗糖酶活性。
28.权利要求27的DNA，该DNA为以下(a6)或者(b6)中所定义的DNA
(a6)一种DNA，该DNA具有序列表中SEQ ID NO93的核苷酸序列，
(b6)一种DNA，该DNA同序列表中的SEQ ID NO93的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有蔗糖酶活性的蛋白质。
29.一种DNA，该DNA编码的蛋白质具有SEQ ID NOS17-20的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NOS17-20的氨基酸序列，并且该蛋白质具有参与谷氨酸摄入的功能。
30.权利要求29的DNA，该DNA为以下(a7)或者(b7)中所定义的DNA
(a7)一种DNA，该DNA具有序列表中SEQ ID NO16的核苷酸序列，
(b7)一种DNA，该DNA同序列表中的SEQ ID NO16的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有参与谷氨酸摄入的功能的蛋白质。
31.一种DNA，该DNA编码的蛋白质具有SEQ ID NO22的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO22的氨基酸序列，并且该蛋白质具有丙酮酸脱氢酶活性。
32.权利要求31的DNA，该DNA为以下(a8)或者(b8)中所定义的DNA
(a8)一种DNA，该DNA具有序列表中SEQ ID NO21的核苷酸序列，
(b8)一种DNA，该DNA同序列表中的SEQ ID NO21的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有丙酮酸脱氢酶活性的蛋白质。
33.一种DNA，该DNA编码的蛋白质具有SEQ ID NO24的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO24的氨基酸序列，并且该蛋白质具有丙酮酸羧化酶活性。
34.权利要求33的DNA，该DNA为以下(a9)或者(b9)中所定义的DNA
(a9)一种DNA，该DNA具有序列表中SEQ ID NO23的核苷酸序列，
(b9)一种DNA，该DNA同序列表中的SEQ ID NO23的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有丙酮酸羧化酶活性的蛋白质。
35.一种DNA，该DNA编码的蛋白质具有SEQ ID NO26的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO26的氨基酸序列，并且该蛋白质具有磷酸烯醇式丙酮酸羧化酶活性。
36.权利要求35的DNA，该DNA为以下(a10)或者(b10)中所定义的DNA
(a10)一种DNA，该DNA具有序列表中SEQ ID NO25的核苷酸序列，
(b10)一种DNA，该DNA同序列表中的SEQ ID NO25的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有磷酸烯醇式丙酮酸羧化酶活性的蛋白质。
37.一种DNA，该DNA编码的蛋白质具有SEQ ID NO28的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO28的氨基酸序列，并且该蛋白质具有顺乌头酸酶活性。
38.权利要求37的DNA，该DNA为以下(a11)或者(b11)中所定义的DNA
(a11)一种DNA，该DNA具有序列表中SEQ ID NO27的核苷酸序列，
(b11)一种DNA，该DNA同序列表中的SEQ ID NO27的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有顺乌头酸酶活性的蛋白质。
39.一种DNA，该DNA编码的蛋白质具有SEQ ID NO30的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO30的氨基酸序列，并且该蛋白质具有异柠檬酸脱氢酶活性。
40.权利要求39的DNA，该DNA为以下(a12)或者(b12)中所定义的DNA
(a12)一种DNA，该DNA具有序列表中SEQ ID NO27的核苷酸序列，
(b12)一种DNA，该DNA同序列表中的SEQ ID NO27的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有异柠檬酸脱氢酶活性的蛋白质。
41.一种DNA，该DNA编码的蛋白质具有SEQ ID NO32的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO32的氨基酸序列，并且该蛋白质具有二氢硫辛酰胺脱氢酶活性。
42.权利要求41的DNA，该DNA为以下(a13)或者(b13)中所定义的DNA
(a13)一种DNA，该DNA具有序列表中SEQ ID NO31的核苷酸序列，
(b13)一种DNA，该DNA同序列表中的SEQ ID NO31的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有二氢硫辛酰胺脱氢酶活性的蛋白质。
43.一种DNA，该DNA编码的蛋白质具有SEQ ID NO34的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO34的氨基酸序列，并且该蛋白质具有2-酮戊二酸脱氢酶活性。
44.权利要求43的DNA，该DNA为以下(a14)或者(b14)中所定义的DNA
(a14)一种DNA，该DNA具有序列表中SEQ ID NO33的核苷酸序列，
(b14)一种DNA，该DNA同序列表中的SEQ ID NO33的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码一种具有2-酮戊二酸脱氢酶活性的蛋白质。
45.一种DNA，该DNA编码的蛋白质具有SEQ ID NO80的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO80的氨基酸序列，并且该蛋白质在42℃下所表现出的谷氨酸脱氢酶活性与37℃下的活性相同或者比后者更高。
46.权利要求45的DNA，该DNA为以下(a15)或者(b15)中所定义的DNA
(a15)一种DNA，该DNA具有序列表中SEQ ID NO79的核苷酸序列，
(b15)一种DNA，该DNA同序列表中的SEQ ID NO79的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码的蛋白质在42℃下所表现出的谷氨酸脱氢酶活性与37℃下的活性相同或者比后者更高。
47.一种DNA，该DNA编码的蛋白质具有SEQ ID NO90的氨基酸序列或者具有包含有一个或者多个氨基酸残基的替换、缺失、插入、添加或者反转的SEQ ID NO90的氨基酸序列，并且该蛋白质在37℃下所表现出的柠檬酸合成酶活性与23℃下的活性相同或者比后者更高。
48.权利要求47的DNA，该DNA为以下(a16)或者(b16)中所定义的DNA
(a16)一种DNA，该DNA具有序列表中SEQ ID NO89的核苷酸序列，
(b16)一种DNA，该DNA同序列表中的SEQ ID NO89的核苷酸序列或者同基于该核苷酸序列而制备的引物可在严格的条件下杂交，并且该DNA编码的蛋白质在37℃下所表现出的柠檬酸合成酶活性与23℃下的活性相同或者比后者更高。
49.一种用于生产L-氨基酸的方法，该方法包括在一种培养基中培养一种微生物，在该培养基中产生并且积累L-氨基酸，以及从该培养基中收集该L-氨基酸，该微生物中引入了权利要求17到48中任意一种的DNA。
全文摘要
基于在多种微生物中所观察到的已知基因序列在氨基酸水平的保守区域，设计了一些复合引物组合，该已知基因对应于一种来自于Corynebacterium thermoaminogenes的编码L－氨基酸生物合成途径的酶的基因，在优选的情况下该酶在比谷氨酸棒状杆菌更高的温度下发挥功能。通过使用这些引物并且使用Corynebacterium thermoaminogenes的染色体DNA作为模板进行了PCR。可用于获得扩增片段的引物被用作为引物，以进行从Corynebacterium thermoaminogenes的染色体DNA质粒文库中选择含有一种靶DNA片段的克隆的筛选。
文档编号C12N9/88GK1391612SQ0081608
公开日2003年1月15日 申请日期2000年10月4日 优先权日1999年10月4日
发明者平野圣子, 野中源, 松崎友美, 秋好直树, 中村佳苗, 木村英一郎, 大住刚, 松井和彦, 河原义雄, 仓桥修, 中松亘, 杉本慎一 申请人:味之素株式会社