一种依赖烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的复合乳酸杆菌短链脱氢酶/还原酶：特性及在（R）-1-苯乙醇合成中的应用
An NADPH-dependent Lactobacillus composti short-chain dehydrogenase/reductase: characterization and application to (R)-1-phenylethanol synthesis.
作者信息
Wang Ya-Jun, Ying Bin-Bin, Chen Min, Shen Wei, Liu Zhi-Qiang, Zheng Yu-Guo
机构信息
Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, People's Republic of China.
Engineering Research Center of Bioconversion and Biopurification of the Ministry of Education, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, People's Republic of China.
出版信息
World J Microbiol Biotechnol. 2017 Jul;33(7):144. doi: 10.1007/s11274-017-2311-9. Epub 2017 Jun 16.
A new anti-Prelog short-chain dehydrogenase/reductase (SDR) encoding gene lcsdr was cloned from Lactobacillus composti DSM 18527, and heterologously expressed in Escherichia coli. LcSDR is nicotinamide adenine dinucleotide phosphate (NADPH)-dependent and has a molecular weight of approximately 30 kDa. The optimal pH and temperature were 6.5 and 30 °C, respectively. The maximal reaction rate V was 133.9 U mg; the Michaelis-Menten constant K of LcSDR were 0.345 mM for acetophenone (1a), and 0.085 mM for NADPH. Through introducing an EsGDH-catalyzed NADPH regeneration system, a biocatalytic process for (R)-1-phenylethanol ((R)-1b) was developed with outstanding time-space yield. Under the optimized conditions, 50 g l 1a was converted to (R)-1b in 2 h with a yield of 93.8%, enantiomeric excess of product (e.e.) above 99% and space-time yield of 562.8 g l d.
从堆肥乳杆菌DSM 18527中克隆出一个新的编码抗普雷洛格短链脱氢酶/还原酶（SDR）的基因lcsdr，并在大肠杆菌中进行了异源表达。LcSDR依赖于烟酰胺腺嘌呤二核苷酸磷酸（NADPH），分子量约为30 kDa。最佳pH值和温度分别为6.5和30℃。最大反应速率V为133.9 U mg；LcSDR对苯乙酮（1a）的米氏常数K为0.345 mM，对NADPH的米氏常数K为0.085 mM。通过引入EsGDH催化的NADPH再生系统，开发了一种用于生产（R）-1-苯乙醇（（R）-1b）的生物催化工艺，具有出色的时空产率。在优化条件下，50 g l 1a在2小时内转化为（R）-1b，产率为93.8%，产物对映体过量（e.e.）高于99%，时空产率为562.8 g l d。
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