title: Isolation and partial characterisation of ACV synthetase from Cephalosporium acremonium and Streptomyces clavuligerus. Evidence for the presence of phosphopantothenate in ACV synthetase.
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.
authors.affiliation: Dyson Perrins Laboratory, University of Oxford, UK.
authors.forename: J E
authors.initials: JE
authors.lastname: Baldwin
authors.forename: J W
authors.initials: JW
authors.lastname: Bird
authors.forename: R A
authors.initials: RA
authors.lastname: Field
authors.forename: N M
authors.initials: NM
authors.lastname: O'Callaghan
authors.forename: C J
authors.initials: CJ
authors.lastname: Schofield
authors.forename: A C
authors.initials: AC
authors.lastname: Willis
mesh_terms: D000164:Acremonium / Q000201:enzymology*; D000595:Amino Acid Sequence; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010205:Pantothenic Acid / Q000031:analogs & derivatives* / Q000737:chemistry; D010453:Peptide Synthases / Q000737:chemistry* / Q000302:isolation & purification; D013302:Streptomyces / Q000201:enzymology*
chemical_list: D010205:Pantothenic Acid; C011988:phosphopantothenic acid; D010453:Peptide Synthases; C095368:alpha-aminoadipyl-cysteinyl-valine synthetase