A genomic search approach to identify carbonyl reductases in Gluconobacter oxydans for enantioselective reduction of ketones
A genomic search approach to identify carbonyl reductases in Gluconobacter oxydans for enantioselective reduction of ketones
复制标题
一种基因组搜索方法，用于鉴定氧化葡糖杆菌中用于对映选择性还原酮的羰基还原酶
DOI：
10.1080/09168451.2014.925775
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发表时间：
2014-08-01
影响因子：
1.6
通讯作者：
Wei, Dongzhi
中科院分区：
文献类型：
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作者：
Chen, Rong;Liu, Xu;Wei, Dongzhi
利用基因组搜索技术，对葡萄糖酸氧化酶中的羰基还原酶进行了研究。所有纯化的酶对测试的酮酯具有不同的活性。用NAD（P）H原位再生体系还原4-苯基-2-丁酮，得到（S）-醇，其e. e.由Gox 0644催化的高达100%。在相同的实验条件下，所有酶都能催化4-氯乙酰乙酸乙酯合成手性产物，具有很好的e. e。99%，除了Gox 0644。Gox 2036对辅酶NADH有严格的要求，在（R）-4-氯-3-羟基丁酸乙酯的合成中表现出良好的对映体专一性。对2-氧代-4-苯基丁酸乙酯的还原反应，具有良好的e. e. Gox 0525获得了较高的转化率（>99%）和较高的转化率（93.1%），而其他酶表现出相对较低的e.和转换。其中Gox 2036和Gox 0525在手性醇的合成中显示出潜在的生物催化活性。图形摘要利用基因组搜索技术研究了葡萄糖酸氧化酶中的羰基还原酶对三种酮的不对称还原反应
The versatile carbonyl reductases from Gluconobacter oxydans in the enantioselective reduction of ketones to the corresponding alcohols were exploited by genome search approach. All purified enzymes showed activities toward the tested ketoesters with different activities. In the reduction of 4-phenyl-2-butanone with in situ NAD(P)H regeneration system, (S)-alcohol was obtained with an e.e. of up to 100% catalyzed by Gox0644. Under the same experimental condition, all enzymes catalyzed ethyl 4-chloroacetoacetate to give chiral products with an excellent e.e. of up to 99%, except Gox0644. Gox2036 had a strict requirement for NADH as the cofactor and showed excellent enantiospecificity in the synthesis of ethyl (R)-4-chloro-3-hydroxybutanoate. For the reduction of ethyl 2-oxo-4-phenylbutyrate, excellent e.e. (>99%) and high conversion (93.1%) were obtained by Gox0525, whereas the other enzymes showed relatively lower e.e. and conversions. Among them, Gox2036 and Gox0525 showed potentials in the synthesis of chiral alcohols as useful biocatalysts. Graphical Abstract The carbonyl reductases from Gluconobacter oxydans exploited by genome search approach were investigated the enantioselective reduction of three ketones