United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima, Kagoshima, 890-8580, Japan.
J Ind Microbiol Biotechnol. 2012 Jan;39(1):55-62. doi: 10.1007/s10295-011-0998-4. Epub 2011 Jun 17.
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.
一株产热不稳定酚酸脱羧酶的从毕赤酵母属假丝酵母（毕赤酵母的无性型）被通过简单连续柱层析在 3 天内纯化为均相。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定的分子量为 20 kDa，用凝胶过滤层析测定的分子量为 36 kDa，表明纯化的酶是一个同二聚体。最适 pH 和温度约为 6.0 和 25°C。该酶的特点是在 0°C 时表现出超过 50%的最适活性，表明该酶具有冷适应特性。该酶将对香豆酸、阿魏酸和咖啡酸转化为相应的产物，其比活约为 600、530 和 46 U/mg。该酶的活性被 Mg2+离子刺激，但被 Fe2+、Ni2+、Cu2+、Hg2+、4-氯汞苯甲酸、N-溴代丁二酰亚胺和二乙基焦碳酸酯完全抑制。该酶可诱导，在细胞内中度由阿魏酸和对香豆酸表达，在非代谢性 6-羟基-2-萘甲酸的作用下显著表达。
