Efficient expression in E. coli of an enantioselective nitrile hydratase from Rhodococcus erythropolis

Liya Song, Hong Jie Yuan, Lee Coffey, John Doran, Mei Xiang Wang, Shijun Qian, Catherine O'Reilly

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

The genes encoding an enantioselective nitrile hydratase (NHase) from Rhodococcus erythropolis AJ270 have been cloned and an active NHase has been produced in Escherichia coli. Maximal activity was found when the genes encoding the α- and β-subunits were transcribed as one unit and the gene encoding the P44k activator protein as a separate ORF on a single replicon. Addition of n-butyric acid and FeSO4 could improve NHase activity. Coexpression of the GroEL-GroES chaperone proteins increased activity in the absence of P44k protein but had no effect in the presence of P44k. The recombinant enzyme was highly enantioselective in the synthesis of S-(+)-3-benzoyloxy- 4-cyanobutyramide from the prochiral substrate 3-benzoyloxyglutaronitrile.

Original languageEnglish
Pages (from-to)755-762
Number of pages8
JournalBiotechnology Letters
Volume30
Issue number4
DOIs
Publication statusPublished - Apr 2008

Keywords

  • Activator protein
  • Enantioselectivity
  • GroES-GroEL
  • Nitrile hydratase
  • Rhodococcus erythropolis

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