Isolation and Partial Characterisation of ACV Synthetase From Cephalosporium Acremonium and Streptomyces Clavuligerus. Evidence for the Presence of Phosphopantothenate in ACV Synthetase

J Antibiot (Tokyo). 1991 Feb;44(2):241-8. doi: 10.7164/antibiotics.44.241.

Abstract

delta-(L-alpha-Aminoadipoyl)-L-cysteinyl-D-valine (ACV) synthetase was isolated and partially characterised from Cephalosporium acremonium CO728 and Streptomyces clavuligerus. The purification procedure resulted in a 745- and 277-fold increase in specific enzyme activity, respectively. Both enzymes had similar apparent molecular masses of ca. 300 kdaltons by SDS-polyacrylamide electrophoresis, under reducing and denaturing conditions, and in excess of 600 kdaltons in the native state by gel filtration. Attempts to obtain an N-terminal amino acid sequence of ACV synthetase from C. acremonium were unsuccessful, hence internal amino acid sequence data were obtained after tryptic digestion of the protein. Phosphopantothenic acid was shown to be associated with the enzyme from both sources, which suggests the possible involvement of pantothenate as a 'swinging arm' in the formation of the tripeptide ACV.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acremonium / enzymology*
  • Amino Acid Sequence
  • Chemical Fractionation
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Pantothenic Acid / analogs & derivatives*
  • Pantothenic Acid / chemistry
  • Peptide Synthases / chemistry*
  • Peptide Synthases / isolation & purification
  • Streptomyces / enzymology*

Substances

  • Pantothenic Acid
  • phosphopantothenic acid
  • Peptide Synthases
  • alpha-aminoadipyl-cysteinyl-valine synthetase