title: Purification and characterization of ferulate and p-coumarate decarboxylase from Bacillus pumilus.
abstract: Bacillus pumilus PS213 isolated from bovine ruminal fluid was able to transform ferulic acid and p-coumaric acid to 4-vinylguaiacol and 4-vinylphenol, respectively, by nonoxidative decarboxylation. The enzyme responsible for this activity has been purified and characterized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of crude extract from a culture induced by ferulic acid or p-coumaric acid shows three bands that are not present in the crude extract of an uninduced culture, while the purified enzyme shows a single band of 23 kDa; the molecular mass calculated by size exclusion chromatography is 45 kDa. Enzyme activity is optimal at 37 degrees C and pH 5.5 and is not enhanced by any cation. Kinetic studies indicated a Km of 1.03 mM and a Vmax of 0.19 mmol.min-1/mg.liter-1 for ferulic acid and a Km of 1.38 mM and a Vmax of 0.22 mmol.min-1/mg.liter-1 for p-coumaric acid.
authors.affiliation: Microbiology Department, International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Degrassi
authors.forename: P
authors.initials: P
authors.lastname: Polverino De Laureto
authors.forename: C V
authors.initials: CV
authors.lastname: Bruschi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000201:enzymology*; D002262:Carboxy-Lyases / Q000302:isolation & purification*; D002417:Cattle; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight
chemical_list: D002262:Carboxy-Lyases; C087194:phenylacrylic acid decarboxylase