A mutant Bacillus subtilis gamma-glutamyltranspeptidase specialized in hydrolysis activity

FEMS Microbiol Lett. 2003 Jul 29;224(2):169-73. doi: 10.1016/S0378-1097(03)00456-7.

Abstract

gamma-Glutamyltranspeptidase (GGT) catalyzes the hydrolysis of gamma-glutamyl compounds and the transfer of their gamma-glutamyl moieties to amino acids and peptides. The transpeptidation activity of Bacillus subtilis GGT is about 10-fold higher than its hydrolysis activity. In B. subtilis GGT, substitution of Asp-445 with Ala abolished its transpeptidation activity. The specific activity for hydrolysis of D445A GGT was 40.2% of that of the wild-type GGT. The K(m) value for L-glutamine was 15.3 mM. D445A GGT was salt tolerant like the wild-type GGT. These results indicate that D445A GGT will be highly useful as a 'glutaminase' in food industry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / enzymology*
  • Bacillus subtilis / genetics
  • Enzyme Activation / drug effects
  • Escherichia coli
  • Glutaminase / metabolism
  • Hydrolysis
  • Kinetics
  • Mutagenesis, Site-Directed
  • Sodium Chloride / pharmacology
  • gamma-Glutamyltransferase / genetics*
  • gamma-Glutamyltransferase / isolation & purification
  • gamma-Glutamyltransferase / metabolism*

Substances

  • Sodium Chloride
  • gamma-Glutamyltransferase
  • Glutaminase