title: Purification and properties of phenolic acid decarboxylase from Candida guilliermondii.
abstract: A heat-labile phenolic acid decarboxylase from Candida guilliermondii (an anamorph of Pichia guilliermondii) was purified to homogeneity by simple successive column chromatography within 3 days. The molecular mass was 20 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 36 kDa by gel-filtration chromatography, suggesting that the purified enzyme is a homodimer. The optimal pH and temperature were approximately 6.0 and 25°C. Characteristically, more than 50% of the optimal activity was observed at 0°C, suggesting that this enzyme is cold-adapted. The enzyme converted p-coumaric acid, ferulic acid, and caffeic acid to corresponding products with high specific activities of approximately 600, 530, and 46 U/mg, respectively. The activity was stimulated by Mg(2+) ions, whereas it was completely inhibited by Fe(2+), Ni(2+), Cu(2+), Hg(2+), 4-chloromericuribenzoate, N-bromosuccinimide, and diethyl pyrocarbonate. The enzyme was inducible and expressed inside the cells moderately by ferulic acid and p-coumaric acid and significantly by non-metabolizable 6-hydroxy-2-naphthoic acid.
authors.affiliation: United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima, Kagoshima, 890-8580, Japan.
authors.forename: Hui-Kai
authors.initials: HK
authors.lastname: Huang
authors.forename: Masamichi
authors.initials: M
authors.lastname: Tokashiki
authors.forename: Sayaka
authors.initials: S
authors.lastname: Maeno
authors.forename: Shoko
authors.initials: S
authors.lastname: Onaga
authors.forename: Toki
authors.initials: T
authors.lastname: Taira
authors.forename: Susumu
authors.initials: S
authors.lastname: Ito
mesh_terms: D002109:Caffeic Acids / Q000378:metabolism; D002175:Candida / Q000201:enzymology*; D002262:Carboxy-Lyases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D002850:Chromatography, Gel; D003373:Coumaric Acids / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D009281:Naphthalenes / Q000378:metabolism; D011422:Propionates; D013379:Substrate Specificity
chemical_list: D002109:Caffeic Acids; D003373:Coumaric Acids; D008670:Metals; D009281:Naphthalenes; D011422:Propionates; C004999:ferulic acid; D002262:Carboxy-Lyases; C417714:phenolic acid decarboxylase; C495469:p-coumaric acid; C030037:2-naphthoic acid; C040048:caffeic acid