发明名称 |
THERMOSTABLE ALCOHOL DEHYDROGENASE DERIVED FROM THERMOCOCCUS GUAYMASENSIS |
摘要 |
An alcohol dehydrogenase (ADH) from hyperthermophilic archaeon Thermococcus guaymasensis was purified to homogeneity and was found to be a homotetramer with a subunit size of 40±1 kDa. The gene encoding the enzyme was cloned and sequenced, and found to have significant sequence homology to known zinc-containing ADHs and L-threonine dehydrogenases with both binding motifs of catalytic zinc and NADP+. The wild-type enzyme is a primary-secondary ADH that exhibits a substrate preference for secondary alcohols and corresponding ketones, and exhibits unusual stereoselectivity. The wild-type enzyme was found to have outstanding thermostability, demonstrating 60% activity after incubation at 80° C. for 40 hours. Site-directed mutagenesis was used to substitute the cyteine residue at position 56 with a serine, to provide the TgADH(C56S) mutant. In the assays that we carried out, we found virtually no difference in enzyme activity and oxygen-sensitivity between the mutant TgADH (C56S) and wild type TgADH. |
申请公布号 |
EP2346992(A1) |
申请公布日期 |
2011.07.27 |
申请号 |
EP20090815527 |
申请日期 |
2009.09.25 |
申请人 |
MA, KESEN;YING, XIANGXIAN |
发明人 |
MA, KESEN;YING, XIANGXIAN |
分类号 |
C12N9/04;C07H21/04;C12N1/21;C12N15/53 |
主分类号 |
C12N9/04 |
代理机构 |
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代理人 |
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地址 |
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