【摘要】 根据纯化得到的(R)-专一性羰基还原酶(rCR)蛋白质测序结果推导出的核苷酸序列设计引物,以筛选得到的近平滑假丝酵母(Candida parapsilosis)CCTCC M203011基因组为模板,通过PCR扩增目的片段,克隆后测序。核苷酸序列测定结果表明rcr基因全长1011bp,共编码336个氨基酸,分子量为35·9kD。将序列递交NCBI比对,与醇脱氢酶超家族成员序列同源性达99%。在大肠杆菌(Escherichia coli)JM109中表达rcr基因,重组菌可还原β-羟基苯乙酮得到(R)-苯乙二醇,光学纯度为100%e·e,摩尔产率为80·4%,在反应体系中无需外加辅酶再生系统即可完成转化。更多还原

【Abstract】 The gene which encodes (R)-specific carbonyl reductase(rCR) from Candida parapsilosis CCTCC M203011 was cloned.The cloned sequence includes an open reading frame (ORF) consisting of 1011bp, encoding a protein of 336 amino acids, with a molecular weight of 35.9 kD.The nucleotide sequence showed 99% similarity to those of the other members of the alcohol dehydrogenase superfamily.A recombinant Escherichia coli JM109 strain harboring the expression plasmid, produced (R)-1-phenyl-1,2-ethanediol(100%e.e,80.4%yield) from β-hydroxyacetophenone without any additive to regenerate NAD~ + from NADH.更多还原