Disclosed is an alcohol dehydrogenase mutant and application thereof in cofactor regeneration, and belongs to the technical fields of enzyme engineering and bioengineering. The alcohol dehydrogenase mutant is obtained by mutating valine at position 84 and/or tyrosine at position 127 in alcohol dehydrogenase having an original amino acid sequence as set forth in SEQ ID No. 1. The alcohol dehydrogenase mutant has high activity for a variety of alcohol co-substrates, and can catalyze these enzyme co-substrates for the regeneration of cofactor NADPH. Compared with the wild-type alcohol dehydrogenase KpADH, the alcohol dehydrogenase mutant has higher activity and catalytic efficiency, and for co-substrate 1,4-butanediol, its kcat value can be up to 75.9 min−1, its kcat/Km value can be up to 2009 min−1·M−1, and its Km value can be as low as 11.3 mM. Therefore, the alcohol dehydrogenase mutant has a higher value in industrial application.
本发明公开了一种醇脱氢酶突变体及其在辅助因子再生中的应用,属于酶工程和
生物工程技术领域。该醇脱氢酶突变体是通过突变醇脱氢酶中 84 位的缬
氨酸和/或 127 位的
酪氨酸而获得的,其原始
氨基酸序列如
SEQ ID No.该醇脱氢酶突变体对多种
醇类共底物具有高活性,并能催化这些酶的共底物以再生辅助因子
NADPH。与野生型醇脱氢酶 KpADH 相比,醇脱氢酶突变体具有更高的活性和催化效率,对于共底物
1,4-丁二醇,其 kcat 值可达 75.9 min-1,kcat/Km 值可达 2009 min-1-M-1,Km 值可低至 11.3 mM。因此,醇脱氢酶突变体具有更高的工业应用价值。