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

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Abstract

(R)-ethyl-3-hydroxyglutarate with highly optical purity (≥99%) 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 β-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 MgSO4·7H2O per liter at 30°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, 30°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 μmol min−1 g−1, respectively.

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Acknowledgements

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).

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Authors and Affiliations

  1. Institute of Bioengineering, Zhejiang University of Technology, Hangzhou, Zhejiang, 310014, People’s Republic of China

    Hua-Ping Dong, Zhi-Qiang Liu, Yu-Guo Zheng & Yin-Chu Shen

  2. Institute of Chemistry and Chemical Engineering, Shaoxing University, Shaoxing, Zhejiang, 312000, People’s Republic of China

    Hua-Ping Dong

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  1. Hua-Ping Dong
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Correspondence to Yu-Guo Zheng.

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Dong, HP., Liu, ZQ., Zheng, YG. et al. Novel biosynthesis of (R)-ethyl-3-hydroxyglutarate with (R)-enantioselective hydrolysis of racemic ethyl 4-cyano-3-hydroxybutyate by Rhodococcus erythropolis . Appl Microbiol Biotechnol 87, 1335–1345 (2010). https://doi.org/10.1007/s00253-010-2584-5

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