文献类型：期刊文献

英文题名：Novel biosynthesis of (R)-ethyl-3-hydroxyglutarate with (R)-enantioselective hydrolysis of racemic ethyl 4-cyano-3-hydroxybutyate by Rhodococcus erythropolis

作者：Dong, Hua-Ping[1,2];Liu, Zhi-Qiang[1];Zheng, Yu-Guo[1];Shen, Yin-Chu[1]

机构：[1]Zhejiang Univ Technol, Inst Bioengn, Hangzhou 310014, Zhejiang, Peoples R China;[2]Shaoxing Univ, Inst Chem & Chem Engn, Shaoxing 312000, Zhejiang, Peoples R China

年份：2010

卷号：87

期号：4

起止页码：1335

外文期刊名：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

收录：SCI-EXPANDED(收录号：WOS:000279200600013)、、EI(收录号：20103313155695)、Scopus(收录号：2-s2.0-77955554529)、WOS

基金：This work was supported by the Major Basic Research Development Program of China (973 Program) (No. 2009CB724704), National High Technology Research and Development Program of China (863 Program) (No. 2009AA02Z203), and the Natural Scientific Foundation of Zhejiang (No. Z4090612).

语种：英文

外文关键词：Rosuvastatin; (R)-ethyl-3-hydroxyglutarate; Enantioselective hydrolysis; Colorimetric screening method; Nitrilase; Rhodococcus erythropolis

外文摘要：(R)-ethyl-3-hydroxyglutarate with highly optical purity (a parts per thousand yen99%) can be used as a novel precursor for synthesis of chiral side chain of rosuvastatin. In this study, a novel synthesis route of (R)-ethyl-3-hydroxyglutarate by whole microorganism cells from racemic ethyl 4-cyano-3-hydroxybutyate was created. A strain ZJB-0910 capable of transforming racemic beta-hydroxy aliphatic nitrile was isolated by employing a screening method based on a colorimetric reaction of Co2+ ion with ammonia, and identified as Rhodococcus erythropolis based on its morphology, physiological tests, Biolog, and the 16S rDNA sequence. After cultivation in a sterilized medium with composition of 20 g glucose, 5 g yeast extract, 0.5 g KH2PO4, 0.5 g K2HPO4, 0.2 g MgSO4 center dot 7H(2)O per liter at 30A degrees C and 150 rpm for 48 h, the whole cells of R. erythropolis ZJB-0910 were prepared as a catalyst in (R)-enantioselective hydrolysis of racemic ethyl 4-cyano-3-hydroxybutyate for synthesis of (R)-ethyl-3-hydroxyglutarate, without bearing hydrolase activity for the ester bond of ethyl 4-cyano-3-hydroxybutyate. Under the optimized biotransformation conditions of pH 7.5, 30A degrees C, and 20 mM substrate concentration, (R)-ethyl-3-hydroxyglutarate with 46.2% yield (ee > 99%) was afforded, and its chemical structure was determined by ESI-MS, NMR, and IR. The apparent Michaelis constant K (m) and maximum rate V (max) for this biocatalytic reaction were 0.01 M and 85.6 mu mol min(-1) g(-1), respectively.