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Efficient production of (R)-o-chloromandelic acid by deracemization of o-chloromandelonitrile with a new nitrilase mined from Labrenzia aggregata

  • Biotechnological products and process engineering
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Abstract

(R)-o-Chloromandelic acid is the key precursor for the synthesis of Clopidogrel®, a best-selling cardiovascular drug. Although nitrilases are often used as an efficient tool in the production of α-hydroxy acids, there is no practical nitrilase specifically developed for (R)-o-chloromandelic acid. In this work, a new nitrilase from Labrenzia aggregata (LaN) was discovered for the first time by genomic data mining, which hydrolyzed o-chloromandelonitrile with high enantioselectivity, yielding (R)-o-chloromandelic acid in 96.5% ee. The LaN was overexpressed in Escherichia coli BL21 (DE3), purified, and its catalytic properties were studied. When o-chloromandelonitrile was used as the substrate, the V max and K m of LaN were 2.53 μmol min−1 mg−1 protein and 0.39 mM, respectively, indicating its high catalytic efficiency. In addition, a study of substrate spectrum showed that LaN prefers to hydrolyze arylacetonitriles. To relieve the substrate inhibition and to improve the productivity of LaN, a biphasic system of toluene–water (1:9, v/v) was adopted, in which o-chloromandelonitrile of 300 mM (apparent concentration, based on total volume) could be transformed by LaN in 8 h, giving an isolated yield of 94.5%. The development of LaN makes it possible to produce (R)-o-chloromandelic acid by deracemizing o-chloromandelonitrile with good ee value and high substrate concentration.

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Acknowledgments

This work was financially supported by National Natural Science Foundation of China (grant number 20902023), Ministry of Science and Technology, People’s Republic of China (numbers 2011CB710800, 2011AA02A210), Shanghai Committee of Science and Technology (grant number 11431921600), Innovation Program of Shanghai Municipal Education Commission (grant number 11CXY24), and China National Special Fund for State Key Laboratory of Bioreactor Engineering (grant number 2060204).

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Authors and Affiliations

  1. Laboratory of Biocatalysis and Synthetic Biotechnology, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, People’s Republic of China

    Chen-Sheng Zhang, Zhi-Jun Zhang, Chun-Xiu Li, Hui-Lei Yu, Gao-Wei Zheng & Jian-He Xu

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  1. Chen-Sheng Zhang
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  2. Zhi-Jun Zhang
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  3. Chun-Xiu Li
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Correspondence to Gao-Wei Zheng or Jian-He Xu.

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Zhang, CS., Zhang, ZJ., Li, CX. et al. Efficient production of (R)-o-chloromandelic acid by deracemization of o-chloromandelonitrile with a new nitrilase mined from Labrenzia aggregata . Appl Microbiol Biotechnol 95, 91–99 (2012). https://doi.org/10.1007/s00253-012-3993-4

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  • DOI: https://doi.org/10.1007/s00253-012-3993-4

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