Purification and properties of phenolic acid decarboxylase from Candida guilliermondii

• Published in: Journal of industrial microbiology & biotechnology 2012, Vol.39 (1), p.55-62
• Main Authors: Huang, Hui-Kai, Tokashiki, Masamichi, Maeno, Sayaka, Onaga, Shoko, Taira, Toki, Ito, Susumu
• Format: Article
• Language: English
• Subjects: Acids; Antimicrobial agents; Biochemistry; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; caffeic acid; Caffeic Acids - metabolism; Candida - enzymology; Candida guilliermondii; Carboxy-Lyases - chemistry; Carboxy-Lyases - isolation & purification; Carboxy-Lyases - metabolism; Chromatography; Chromatography, Gel; Coumaric Acids - metabolism; Electrophoresis, Polyacrylamide Gel; Enzymes; Experiments; ferulic acid; Fundamental and applied biological sciences. Psychology; gel electrophoresis; Gene expression; Genetic Engineering; Glucose; Inorganic Chemistry; ions; Life Sciences; magnesium; mercury; Metals - pharmacology; Meyerozyma guilliermondii; Microbiology; Molecular Weight; Naphthalenes - metabolism; Original Paper; p-coumaric acid; Phenols; Pichia guilliermondii; Propionates; Proteins; Sensors; Sodium; Studies; Substrate Specificity; temperature; Yeast
• ISSN: 1367-5435; 1476-5535
• DOI: 10.1007/s10295-011-0998-4

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 Mg2+ ions, whereas it was completely inhibited by Fe2+, Ni2+, Cu2+, Hg2+, 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.