一种青霉素g酰化酶突变体的制作方法_2

文档序号:9722643阅读:来源:国知局
AALPAADRQVLDGYAAGMNAWLARVRAQPGQL MPKEFNDLGFAPADWTAYDVAMIFVGTMANRASDANSEIDNLALLTALKDRHGAADAMRIFNQLRWLTDSRAPTTVP AEAGSYQPPVFQPDGADPLAYALPRYDGTPPMLERVVRDPATRGVVDGAPATLRAQLAAQYAQSGQPGIAGFPTTSN MWIVGRDHAKDARSILLNGPQFGffffNPAYTYGIGLHGAGFDVVGNTPFAYPSILFGHNAHVTWGSTAGFGDDVDIFA EKLDPADRTRYFHDGQWKTLEKRTDLILVKDAAPVTLDVYRSVHGLIVKFDDAQHVAYAKARAWEGYELQSLMAWTR KTQSANWEQWKAQAARHALTINWYYADDRGNIGYAHTGFYPRRRPGHDPRLPVPGTGEMDWLGLLPFSTNPQVYNPR QGFIANWNNQPMRGYPSTDLFAIVWGQADRYAEIETRLKAMTANGGKVSAQQMWDLIRTTSYADVNRRHFLPFLQRA VQGLPADDPRVRLVAGLAAWDGMMTSERQPGYFDNAGPAVMDAWLRAMLRRTLADEMPADFFKWYSATGYPTPQAPA TGSLNLTTGVKVLFNALAGPEAGVPQRYDFFNGARADDVILAALDDALAALRQAYGQDPAAWKIPAPPMVFAPKNFL GVPQADAKAVLCYRATQNRGTENNMTVFDGKSVRAVDVVAPGQSGFVAPDGTPSPHTRDQFDLYNTFGSKRVWFTAD EVRRNATSEETLRYPR;
[0022] SEQ ID NO:6,其为SEQ ID NO: 1第232位的A替换为E的突变体,其氨基酸序列为:
[0023] MKQQWLSAALLAASSCLPAMAAQPVAPAAGQTSEAVAARPQTADGKVTIRRDAYGMPHVYADTVYGIFYGYGYAVAQ DRLFQMEMARRSTQGRVAEVLGASMVGFDKSIRANFSPERIQRQLAALPAADRQVLDGYAAGMNAWLARVRAQPGQL MPKEFNDLGFAPADWTAYDVAMIFVGTMANRFSDANSEIDNLALLTALKDRHGAADAMRIFNQLRWLTDSRAPTTVP EEAGSYQPPVFQPDGADPLAYALPRYDGTPPMLERVVRDPATRGVVDGAPATLRAQLAAQYAQSGQPGIAGFPTTSN MWIVGRDHAKDARSILLNGPQFGffffNPAYTYGIGLHGAGFDVVGNTPFAYPSILFGHNAHVTWGSTAGFGDDVDIFA EKLDPADRTRYFHDGQWKTLEKRTDLILVKDAAPVTLDVYRSVHGLIVKFDDAQHVAYAKARAWEGYELQSLMAWTR KTQSANWEQWKAQAARHALTINWYYADDRGNIGYAHTGFYPRRRPGHDPRLPVPGTGEMDWLGLLPFSTNPQVYNPR QGFIANWNNQPMRGYPSTDLFAIVWGQADRYAEIETRLKAMTANGGKVSAQQMWDLIRTTSYADVNRRHFLPFLQRA VQGLPADDPRVRLVAGLAAWDGMMTSERQPGYFDNAGPAVMDAWLRAMLRRTLADEMPADFFKWYSATGYPTPQAPA TGSLNLTTGVKVLFNALAGPEAGVPQRYDFFNGARADDVILAALDDALAALRQAYGQDPAAWKIPAPPMVFAPKNFL GVPQADAKAVLCYRATQNRGTENNMTVFDGKSVRAVDVVAPGQSGFVAPDGTPSPHTRDQFDLYNTFGSKRVWFTAD EVRRNATSEETLRYPR;
[0024] SEQ ID NO:7,其为SEQ ID NO: 1第330位的F替换为A的突变体,其氨基酸序列为:
[0025] MKQQWLSAALLAASSCLPAMAAQPVAPAAGQTSEAVAARPQTADGKVTIRRDAYGMPHVYADTVYGIFYGYGYAVAQ DRLFQMEMARRSTQGRVAEVLGASMVGFDKSIRANFSPERIQRQLAALPAADRQVLDGYAAGMNAWLARVRAQPGQL MPKEFNDLGFAPADWTAYDVAMIFVGTMANRFSDANSEIDNLALLTALKDRHGAADAMRIFNQLRWLTDSRAPTTVP AEAGSYQPPVFQPDGADPLAYALPRYDGTPPMLERVVRDPATRGVVDGAPATLRAQLAAQYAQSGQPGIAGFPTTSN MWIVGRDHAKDARSILLNGPQAGffffNPAYTYGIGLHGAGFDVVGNTPFAYPSILFGHNAHVTWGSTAGFGDDVDIFA EKLDPADRTRYFHDGQWKTLEKRTDLILVKDAAPVTLDVYRSVHGLIVKFDDAQHVAYAKARAWEGYELQSLMAWTR KTQSANWEQWKAQAARHALTINWYYADDRGNIGYAHTGFYPRRRPGHDPRLPVPGTGEMDWLGLLPFSTNPQVYNPR QGFIANWNNQPMRGYPSTDLFAIVWGQADRYAEIETRLKAMTANGGKVSAQQMWDLIRTTSYADVNRRHFLPFLQRA VQGLPADDPRVRLVAGLAAWDGMMTSERQPGYFDNAGPAVMDAWLRAMLRRTLADEMPADFFKWYSATGYPTPQAPA TGSLNLTTGVKVLFNALAGPEAGVPQRYDFFNGARADDVILAALDDALAALRQAYGQDPAAWKIPAPPMVFAPKNFL GVPQADAKAVLCYRATQNRGTENNMTVFDGKSVRAVDVVAPGQSGFVAPDGTPSPHTRDQFDLYNTFGSKRVWFTAD EVRRNATSEETLRYPR;
[0026] SEQ ID NO:8,其为SEQ ID NO: 1第415位的K替换为E的突变体,其氨基酸序列为:
[0027] MKQQWLSAALLAASSCLPAMAAQPVAPAAGQTSEAVAARPQTADGKVTIRRDAYGMPHVYADTVYGIFYGYGYAVAQ DRLFQMEMARRSTQGRVAEVLGASMVGFDKSIRANFSPERIQRQLAALPAADRQVLDGYAAGMNAWLARVRAQPGQL MPKEFNDLGFAPADWTAYDVAMIFVGTMANRFSDANSEIDNLALLTALKDRHGAADAMRIFNQLRWLTDSRAPTTVP AEAGSYQPPVFQPDGADPLAYALPRYDGTPPMLERVVRDPATRGVVDGAPATLRAQLAAQYAQSGQPGIAGFPTTSN MWIVGRDHAKDARSILLNGPQFGffffNPAYTYGIGLHGAGFDVVGNTPFAYPSILFGHNAHVTWGSTAGFGDDVDIFA EKLDPADRTRYFHDGQWKTLEKRTDLILVEDAAPVTLDVYRSVHGLIVKFDDAQHVAYAKARAWEGYELQSLMAWTR KTQSANWEQWKAQAARHALTINWYYADDRGNIGYAHTGFYPRRRPGHDPRLPVPGTGEMDWLGLLPFSTNPQVYNPR QGFIANWNNQPMRGYPSTDLFAIVWGQADRYAEIETRLKAMTANGGKVSAQQMWDLIRTTSYADVNRRHFLPFLQRA VQGLPADDPRVRLVAGLAAWDGMMTSERQPGYFDNAGPAVMDAWLRAMLRRTLADEMPADFFKWYSATGYPTPQAPA TGSLNLTTGVKVLFNALAGPEAGVPQRYDFFNGARADDVILAALDDALAALRQAYGQDPAAWKIPAPPMVFAPKNFL GVPQADAKAVLCYRATQNRGTENNMTVFDGKSVRAVDVVAPGQSGFVAPDGTPSPHTRDQFDLYNTFGSKRVWFTAD EVRRNATSEETLRYPR;
[0028] SEQ ID NO:9,其为SEQ ID NO: 1第798位的F替换为L的突变体,其氨基酸序列为:
[0029] MKQQWLSAALLAASSCLPAMAAQPVAPAAGQTSEAVAARPQTADGKVTIRRDAYGMPHVYADTVYGIFYGYGYAVAQ DRLFQMEMARRSTQGRVAEVLGASMVGFDKSIRANFSPERIQRQLAALPAADRQVLDGYAAGMNAWLARVRAQPGQL MPKEFNDLGFAPADWTAYDVAMIFVGTMANRFSDANSEIDNLALLTALKDRHGAADAMRIFNQLRWLTDSRAPTTVP AEAGSYQPPVFQPDGADPLAYALPRYDGTPPMLERVVRDPATRGVVDGAPATLRAQLAAQYAQSGQPGIAGFPTTSN MWIVGRDHAKDARSILLNGPQFGffffNPAYTYGIGLHGAGFDVVGNTPFAYPSILFGHNAHVTWGSTAGFGDDVDIFA EKLDPADRTRYFHDGQWKTLEKRTDLILVKDAAPVTLDVYRSVHGLIVKFDDAQHVAYAKARAWEGYELQSLMAWTR KTQSANWEQWKAQAARHALTINWYYADDRGNIGYAHTGFYPRRRPGHDPRLPVPGTGEMDWLGLLPFSTNPQVYNPR QGFIANWNNQPMRGYPSTDLFAIVWGQADRYAEIETRLKAMTANGGKVSAQQMWDLIRTTSYADVNRRHFLPFLQRA VQGLPADDPRVRLVAGLAAWDGMMTSERQPGYFDNAGPAVMDAWLRAMLRRTLADEMPADFFKWYSATGYPTPQAPA TGSLNLTTGVKVLFNALAGPEAGVPQRYDFFNGARADDVILAALDDALAALRQAYGQDPAAWKIPAPPMVFAPKNFL GVPQADAKAVLCYRATQNRGTENNMTVLDGKSVRAVDVVAPGQSGFVAPDGTPSPHTRDQFDLYNTFGSKRVWFTAD EVRRNATSEETLRYPR;或者
[0030] SEQ ID NO: 10,其为SEQ ID NO: 1第44位的D替换为L、第130位的R替换为M、第186 位的F替换为A、第232位的A替换为E、第330位的F替换为A、第415位的K替换为E、第798位的F 替换为L的突变体,其氨基酸序列为:
[0031] MKQQWLSAALLAASSCLPAMAAQPVAPAAGQTSEAVAARPQTALGKVTIRRDAYGMPHVYADTVYGIFYGYGYAVAQ DRLFQMEMARRSTQGRVAEVLGASMVGFDKSIRANFSPERIQRQLAALPAADMQVLDGYAAGMNAWLARVRAQPGQL MPKEFNDLGFAPADWTAYDVAMIFVGTMANRASDANSEIDNLALLTALKDRHGAADAMRIFNQLRWLTDSRAPTTVP EEAGSYQPPVFQPDGADPLAYALPRYDGTPPMLERVVRDPATRGVVDGAPATLRAQLAAQYAQSGQPGIAGFPTTSN MWIVGRDHAKDARSILLNGPQAGffffNPAYTYGIGLHGAGFDVVGNTPFAYPSILFGHNAHVTWGSTAGFGDDVDIFA EKLDPADRTRYFHDGQWKTLEKRTDLILVEDAAPVTLDVYRSVHGLIVKFDDAQHVAYAKARAWEGYELQSLMAWTR KTQSANWEQWKAQAARHALTINWYYADDRGNIGYAHTGFYPRRRPGHDPRLPVPGTGEMDWLGLLPFSTNPQVY
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