O-acetylhomoserine sulfhydrylase from Clostridium novyi. Cloning, expression of the gene and characterization of the enzyme

Protein Expr Purif. 2021 Apr:180:105810. doi: 10.1016/j.pep.2020.105810. Epub 2020 Dec 16.

Abstract

The gene NT01CX_1210 of pathogenic bacterium Clostridium novyi annotated as encoding O-acetylhomoserine sulfhydrylase was cloned and expressed in Escherichia coli. The gene product having O-acetylhomoserine sulfhydrylase activity was purified to homogeneity. The protein showed molecular mass of approximately 184 kDa for the native form and 46 kDa for the subunit. The enzyme catalyzes the γ-substitution reaction of O-acetylhomoserine with maximum activity at pH 7.5. Analysis of C. novyi genome allowed us to suggest that there is only one way for the synthesis of l-methionine in the bacterium. The data obtained may provide the basis for further study of the role of OAHS in Clostridium bacteria and an ascertainment of its mechanism.

Keywords: Clostridium novyi; Methionine biosynthesis; O-acetylhomoserine sulfhydrylase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins* / biosynthesis
  • Bacterial Proteins* / chemistry
  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / isolation & purification
  • Carbon-Oxygen Lyases* / biosynthesis
  • Carbon-Oxygen Lyases* / chemistry
  • Carbon-Oxygen Lyases* / genetics
  • Carbon-Oxygen Lyases* / isolation & purification
  • Cloning, Molecular*
  • Clostridium / enzymology
  • Clostridium / genetics*
  • Gene Expression*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification

Substances

  • Bacterial Proteins
  • Recombinant Proteins
  • O-acetylhomoserine (thiol)-lyase
  • Carbon-Oxygen Lyases

Supplementary concepts

  • Clostridium novyi