A single amino acid substitution converts gamma-glutamyltranspeptidase to a class IV cephalosporin acylase (glutaryl-7-aminocephalosporanic acid acylase)

Appl Environ Microbiol. 2004 Oct;70(10):6324-8. doi: 10.1128/AEM.70.10.6324-6328.2004.

Abstract

The aspartyl residue at position 433 of gamma-glutamyltranspeptidase of Escherichia coli K-12 was replaced by an asparaginyl residue. This substitution enabled gamma-glutamyltranspeptidase to deacylate glutaryl-7-aminocephalosporanic acid, producing 7-aminocephalosporanic acid, which is a starting material for the synthesis of semisynthetic cephalosporins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Base Sequence
  • DNA, Bacterial / genetics
  • Escherichia coli K12 / enzymology
  • Escherichia coli K12 / genetics
  • Hydrogen-Ion Concentration
  • Magnetic Resonance Spectroscopy
  • Mutagenesis, Site-Directed
  • Penicillin Amidase / chemistry
  • Penicillin Amidase / genetics*
  • Penicillin Amidase / metabolism*
  • Substrate Specificity
  • gamma-Glutamyltransferase / chemistry
  • gamma-Glutamyltransferase / genetics*
  • gamma-Glutamyltransferase / metabolism*

Substances

  • DNA, Bacterial
  • gamma-Glutamyltransferase
  • Penicillin Amidase
  • glutarylamidocephalosporanic acid acylase