NAD+激酶
芳基
酒
马红球菌
荧光
化学
红球菌
醇脱氢酶
醇氧化还原酶
立体化学
有机化学
生物化学
酶
烷基
物理
基因
量子力学
毒力
作者
Chi Yang,Xiangxian Ying,Meilan Yu,Yinjun Zhang,Bin Xiong,Qingqing Song,Zhao Wang
出处
期刊:Journal of Industrial Microbiology & Biotechnology
[Oxford University Press]
日期:2012-10-01
卷期号:39 (10): 1431-1443
被引量:13
标识
DOI:10.1007/s10295-012-1160-7
摘要
Abstract A simple and reliable procedure was developed to screen biocatalysts with high alcohol dehydrogenase activity, efficient internal coenzyme regeneration, and high stereoselectivity. The strategy of activity screening in a microtitre plate format was based on the detection of fluorescence of NAD(P)H originating from the oxidation of alcohols. The primary and secondary screenings from soil samples yielded a versatile bacterial biocatalyst Rhodococcus erythropolis WZ010 demonstrating potential for the preparation of chiral aryl secondary alcohols. In terms of activity and stereoselectivity, the optimized reaction conditions in the stereoselective oxidation were 30 °C, pH 10.5, and 250 rpm, whereas bioreduction using glucose as co-substrate was the most favorable at 35 °C and pH 7.5 in the static reaction mixture. Under the optimized conditions, fresh cells of the strain stereoselectively oxidized the (S)-enantiomer of racemic 1-phenylethanol (120 mM) to acetophenone and afforded the unoxidized (R)-1-phenylethanol in 49.4 % yield and >99.9 % enantiomeric excess (e.e.). In the reduction of 10 mM acetophenone, the addition of 100 mM glucose significantly increased the conversion rate from 3.1 to 97.4 %. In the presence of 800 mM glucose, acetophenone and other aromatic ketones (80 mM) were enantioselectively reduced to corresponding (S)-alcohols with excellent e.e. values. Both stereoselective oxidation and asymmetric reduction required no external cofactor regeneration system.
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