Bacillus D-stereospecific metallo-amidohydrolase: active-site metal-ion substitution changes substrate specificity

Biochimie. 2009 Apr;91(4):568-76. doi: 10.1016/j.biochi.2009.01.015.

Abstract

From investigation of 2000 soil isolates, we identified a D-stereospecific metallo-amidohydrolase that can hydrolyze D-aminoacyl derivatives from the culture supernatant of Bacillus sp. 62E11: 62E11DppA. The enzyme binds two equivalents of zinc, exhibits 70% identity with that of D-aminopeptidases from Bacillus subtilis (DppA). In fact, 62E11DppA has strict specificity toward D-aminoacyl derivatives, i.e., the enzyme shows high activity toward D-aminoacyl benzyl esters and little activity toward D-amino acid containing peptides. Moreover, 62E11DppA exhibits a dramatic change in its activity and substrate specificity by substitution of metal ions in its active site. Based on results of kinetic studies using apo-62E11DppA with various metal ion and substrate concentrations, we propose a possible mechanism for the change in its activity and specificity by substitution of metal ions: the substitution of metal ions in 62E11DppA dramatically changes its activity by altering the substrate specificity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidohydrolases / chemistry
  • Amidohydrolases / isolation & purification
  • Amidohydrolases / metabolism*
  • Amino Acid Sequence
  • Bacillus / enzymology*
  • Catalytic Domain
  • Cations, Divalent / chemistry
  • Cations, Divalent / metabolism
  • Hydrolysis
  • Metals / chemistry
  • Metals / metabolism*
  • Molecular Sequence Data
  • Sequence Alignment
  • Substrate Specificity

Substances

  • Cations, Divalent
  • Metals
  • Amidohydrolases