不对称还原3,5-双三氟甲基苯乙酮的羰基还原酶的挖掘与功能验证

Mining and function verification of carbonyl reductase through 3,5-bistrifluoromethylphenyl ethanol

基因挖掘是获得具有优良性能生物催化剂的新方向。为了获得能不对称催化3,5-双三氟甲基苯乙酮(BTAP)制备(R)-3,5-双三氟甲基苯乙醇(R-BTPE)的羰基还原酶,以文献报道的能不对称催化BTAP生成RBTPE的羰基还原酶LXCAR的氨基酸序列在Gen Bank数据库中进行基因挖掘,获得同源性> 80%的序列24条。从中筛选并合成了同源性为88%、来源于未培养微生物的羰基还原酶BAP47456. 1,在大肠杆菌异源表达并进行功能验证,发现该羰基还原酶对BTAP表现出良好的立体选择性,还原产物为R-BTPE,证明了基因挖掘的有效性。

Gene mining is the new direction of obtaining good biocatalyst. In order to obtain earbonyl reductase of （R） - 3,5 - Bi three fluoromethyl phenyl alcohol （R- BTPE）through 3,5 -bistrifluoromethylphenyl ethanol（BTAP） , we did gene mining of amino acid se-quence of carbonyl reductase of （R） -3,5 - Bi three fluoromethyl phenyl alcohol （R- BTPE）through 3,5 -bistrifluoromethylphenyl ethanol（BTAP） in GenBank, obtaining 24 sequences of homology 〉 80%. The silfted and synthesizd homology is 88%. The carbonyl reductase from the uncultured microorganisms is BAP47456. 1. The functional verification of escherichia coli heterologous shows that carbonyl reductase has good steroselectivity of BTAP and teh reduzate is R-BTPE, procing the effectiveness of gene mining.