Lab of Biocatalysis, Hangzhou Normal University, 402-Building D, 1378 West Wenyi Road, Hangzhou, 311121, China.
Biotechnol Lett. 2013 Mar;35(3):359-65. doi: 10.1007/s10529-012-1082-5. Epub 2012 Nov 18.
The gene encoding a novel short-chain alcohol dehydrogenase in the thermophilic bacterium, Carboxydothermus hydrogenoformans, was identified and overexpressed in Escherichia coli. The enzyme was thermally stable and displayed the highest activity at 70 °C and pH 6.0. It preferred NAD(H) over NADP(H) as a cofactor and exhibited broad substrate specificity towards aliphatic ketones, cycloalkanones, aromatic ketones, and ketoesters. Furthermore, ethyl benzoylformate was asymmetrically reduced by the purified enzyme, using an additional coupled NADH regeneration system, with 95 % conversion and in an enantiomeric excess of (99.9 %). The results of this study may lead to the discovery of a novel method for asymmetric reduction of alcohols, which is an important tool in organic synthesis.
在嗜热细菌产氢羧菌中鉴定并在大肠杆菌中过表达了一种新型短链醇脱氢酶的编码基因。该酶热稳定性好，在 70°C 和 pH6.0 时活性最高。它偏爱 NAD(H)而不是 NADP(H)作为辅助因子，对脂肪族酮、环烷酮、芳香族酮和酮酯具有广泛的底物特异性。此外，在用附加的 NADH 再生系统偶联时，纯化酶可将苯甲酸乙酯不对称还原，转化率为 95%，对映体过量值为（99.9%）。本研究的结果可能会发现一种新型的醇不对称还原方法，这是有机合成中的重要工具。
