PURIFICATION AND CHARACTERIZATION OF FERULATE AND P-COUMARATE DECARBOXYLASE FROM BACILLUS-PUMILUS
PURIFICATION AND CHARACTERIZATION OF FERULATE AND P-COUMARATE DECARBOXYLASE FROM BACILLUS-PUMILUS
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DOI：
10.1128/aem.61.1.326-332.1995
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发表时间：
1995-01-01
影响因子：
4.4
通讯作者：
BRUSCHI, CV
中科院分区：
文献类型：
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作者：
DEGRASSI, G;DELAURETO, PP;BRUSCHI, CV
从牛瘤胃液中分离的矮芽孢杆菌PS213能够通过非氧化脱羧作用将阿魏酸和对香豆酸分别转化为4-乙烯基愈创木酚和4-乙烯基酚。负责这种活性的酶已被纯化和表征。对阿威酸或对香豆酸诱导的粗提物的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示出未诱导的粗提物中不存在的三条条带，而纯化后的酶显示出23 kDa的单一条带；通过尺寸排斥色谱法计算得到的分子质量为45 kDa。酶的活性在37℃和pH 5.5时达到最佳，不受任何阳离子的影响。动力学研究表明，K-m为1.03 mM， V-max为0.19 mmol。阿魏酸的K-m值为1.38 mM， V-max值为0.22 mM .min(-1)/mg。l（-1）表示对香豆酸。
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 K-m of 1.03 mM and a V-max of 0.19 mmol.min(-1)/mg liter(-1) for ferulic acid and a K-m of 1.38 mM and a V-max of 0.22 mmol.min(-1)/mg.liter(-1) for p-coumaric acid.