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Enantioselective synthesis of (S)-2-cyano-2-methylpentanoic acid by nitrilase

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Abstract

The nitrilase gene of Rhodococcus rhodochrous J1 was expressed in Escherichia coli using the expression vector, pKK223-3. The recombinant E. coli JM109 cells hydrolyzed enantioselectively 2-methyl-2-propylmalononitrile to form (S)-2-cyano-2-methylpentanoic acid (CMPA) with 96 % e.e. Under optimized conditions, 80 g (S)-CMPA l−1 was produced with a molar yield of 97 % at 30 °C after a 24 h without any by-products.

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References

  • Bayer S, Birkemeyer C, Ballschmiter M (2011) A nitrilase from a metagenomic library acts regioselectively on aliphatic dinitriles. Appl Microbiol Biotechnol 89:91–98

    Article  PubMed  CAS  Google Scholar 

  • Conway EJ, Berne A (1933) An absorption apparatus for the micro-determination of certain volatile substances. Biochem J 27:419–429

    PubMed  CAS  Google Scholar 

  • Fawcett JK, Scott JE (1960) A rapid and precise method for the determination of urea. J Clin Pathol 13:156–159

    Article  PubMed  CAS  Google Scholar 

  • Kobayashi M, Nagasawa T, Yamada H (1989) Nitrilase of Rhodococcus rhodochrous J1: purification and characterization. Eur J Biochem 182:349–356

    Article  PubMed  CAS  Google Scholar 

  • Kobayashi M, Komeda H, Yanaka N, Nagasawa T, Yamada H (1992) Nitrilase from Rhodococcus rhodochrous J1: sequencing and overexpression of the gene and identification of an essential cysteine residue. J Biol Chem 267:20746–20751

    PubMed  CAS  Google Scholar 

  • Moretto A, Peggion C, Formaggio F, Crisma M, Toniolo C, Piazza C, Kaptein B, Broxterman QB, Ruiz I, Díaz-de-Villegas MD, Galvez JA, Cativiela C (2000) (αMe)Nva: stereoselective syntheses and preferred conformation of selected model peptides. J Peptide Res 56:283–297

    Article  CAS  Google Scholar 

  • Nagasawa T, Yamada H (1989) Microbial transformation of nitriles. Trends Biotechnol 7:153–158

    Article  CAS  Google Scholar 

  • Nagasawa T, Kobayashi M, Yamada H (1988) Optimum culture conditions for the production of benzonitrilase by Rhodococcus rhodochrous J1. Arch Microbiol 150:89–94

    Article  CAS  Google Scholar 

  • Nagasawa T, Wieser M, Nakamura T, Iwahara H, Yoshida T, Gekko K (2000) Nitrilase of Rhodococcus rhodochrous J1: conversion into the active form by subunit association. Eur J Biochem 267:138–144

    Article  PubMed  CAS  Google Scholar 

  • Thuku RN, Weber BW, Varsani A, Sewell BT (2007) Post-translational cleavage of recombinantly expressed nitrilase from Rhodococcus rhodochrous J1 yields a stable, active helical form. FEBS J 274:2099–2108

    Article  PubMed  CAS  Google Scholar 

  • Vejttech V, Sveda O, Kaplan O, Prikrylová V, Elisáková V, Himl M, Kubác D, Pelantová H, Kuzma M, Kren V, Martínková L (2007) Biotransformation of heterocyclic dinitriles by Rhodococcus erythropolis and fungal nitrilases. Biotechnol Lett 29:1119–1124

    Article  Google Scholar 

  • Wu ZL, Li ZY (2003) Enantioselective biotransformation of α,α-disubstituted dinitriles to the corresponding 2-cyanoacetamides using Rhodococcus sp. CGMCC0497. Tetrahedron Asymm 14:2133–2142

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biomolecular Science, Gifu University, Yanagido 1-1, Gifu, 501-1193, Japan

    Toyokazu Yoshida, Koichi Mitsukura, Takuya Mizutani, Ryo Nakashima, Yasuyo Shimizu & Toru Nagasawa

  2. Biotechnology Laboratory, Mitsubishi Chemical Group Science and Technology Research Center, Inc., 1000 Kamoshida-cho, Aoba-ku, Yokohama, 227-8502, Japan

    Hiroshi Kawabata

Authors
  1. Toyokazu Yoshida
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  2. Koichi Mitsukura
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  3. Takuya Mizutani
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  4. Ryo Nakashima
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  5. Yasuyo Shimizu
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  6. Hiroshi Kawabata
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  7. Toru Nagasawa
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Correspondence to Toyokazu Yoshida.

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Yoshida, T., Mitsukura, K., Mizutani, T. et al. Enantioselective synthesis of (S)-2-cyano-2-methylpentanoic acid by nitrilase. Biotechnol Lett 35, 685–688 (2013). https://doi.org/10.1007/s10529-012-1131-0

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  • DOI: https://doi.org/10.1007/s10529-012-1131-0

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