[发明专利]一种耐热碱性磷酸酯酶及其表达

摘要

本发明属生物工程技术领域，涉及一种耐热碱性磷酸酯酶。本发明提供了该耐热碱性磷酸酯酶的氨基酸序列，以及编码该氨基酸序列的DNA片段。本发明还包括克隆有该DNA片段或其一部分的表达载体和生产该重组酶的方法。同时还涉及利用该酶标记生物大分子的方法。

主权项

1.一种耐热碱性磷酸酯酶，其特征在于，含有以下氨基酸序列中的27-501位或1-501位： 1 Met Lys Arg Arg Asp Ile Leu Lys Gly Gly Leu Ala Ala Gly Ala 16 Leu Ala Leu Leu Pro Arg Gly His Thr Gln Gly Ala Leu Gln Asn 31 Gln Pro Ser Leu Gly Arg Arg Tyr Arg Asn Leu Ile Val Phe Val 46 Tyr Asp Gly Phe Ser Trp Glu Asp Tyr Ala Ile Ala Gln Ala Tyr 61 Ala Arg Arg Arg Gln Gly Arg Val Leu Ala Leu Glu Arg Leu Leu 76 Ala Arg Tyr Pro Asn Gly Leu Ile Asn Thr Tyr Ser Leu Thr Ser 91 Tyr Val Thr Glu Ser Ser Ala Ala Gly Asn Ala Phe Ser Cys Gly106 Val Lys Thr Val Asn Gly Gly Leu Ala Ile His Ala Asp Gly Thr121 Pro Leu Lys Pro Phe Phe Ala Ala Ala Lys Glu Ala Gly Lys Ala136 Val Gly Leu Val Thr Thr Thr Thr Val Thr His Ala Thr Pro Ala151 Ser Phe Val Val Ser Asn Pro Asp Arg Asn Ala Glu Glu Arg Ile166 Ala Glu Gln Tyr Leu Glu Phe Gly Ala Glu Val Tyr Leu Gly Gly181 Gly Asp Arg Phe Phe Asn Pro Ala Arg Arg Lys Asp Gly Lys Asp196 Leu Tyr Ala Ala Phe Ala Ala Lys Gly Tyr Gly Val Val Arg Thr211 Pro Glu Glu Leu Ala Arg Ser Asn Ala Thr Arg Leu Leu Gly Val226 Phe Ala Asp Gly His Val Pro Tyr Glu Ile Asp Arg Arg Phe Gln241 Gly Leu Gly Val Pro Ser Leu Lys Glu Met Val Gln Ala Ala Leu256 Pro Arg Leu Ala Ala His Arg Gly Gly Phe Val Leu Gln Val Glu271 Ala Gly Arg Ile Asp His Ala Asn His Leu Asn Asp Ala Gly Ala286 Thr Leu Trp Asp Val Leu Ala Ala Asp Glu Val Leu Glu Leu Leu301 Thr Ala Phe Val Asp Arg Asn Pro Asp Thr Leu Leu Leu Val Val316 Ser Asp His Ala Thr Gly Val Gly Ala Leu Tyr Gly Ala Gly Arg331 Ser Tyr Leu Glu Ser Ser Val Gly Ile Asp Leu Leu Gly Ala Gln346 Lys Ala Ser Phe Glu Tyr Met Arg Arg Val Leu Gly Ser Ala Pro361 Asp Ala Ala Gln Val Lys Glu Ala Tyr Gln Thr Leu Lys Gly Val376 Ser Leu Thr Asp Glu Glu Ala Gln Met Val Val Arg Ala Ile Arg391 Glu Arg Val Tyr Trp Pro Asp Ala Val Arg Gln Gly Ile Gln Pro406 Glu Asn Thr Met Ala Trp Ala Met Val Gln Lys Asn Ala Ser Lys421 Pro Asp Arg Pro Ash Ile Gly Trp Ser Ser Gly Gln His Thr Ala436 Ser Pro Val Ile Leu Leu Leu Tyr Gly Gln Gly Leu Arg Phe Val451 Gln Leu Gly Leu Val Asp Asn Thr His Val Phe Arg Leu Met Gly466 Glu Ala Leu Asn Leu Arg Tyr Gln Asn Pro Val Met Ser Glu Glu481 Glu Ala Leu Glu Ile Leu Lys Ala Arg Pro Gln Gly Met Arg His496 Pro Glu Asp Val Trp Ala。