Purification and characterization of an oxygen-sensitive reversible 4-hydroxybenzoate decarboxylase from Clostridium hydroxybenzoicum.
Purification and characterization of an oxygen-sensitive reversible 4-hydroxybenzoate decarboxylase from Clostridium hydroxybenzoicum.
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来自羟基苯甲酸梭菌的氧敏感可逆 4-羟基苯甲酸脱羧酶的纯化和表征。
DOI：
10.1111/j.1432-1033.1995.0077l.x
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
J. Wiegel
中科院分区：
文献类型：
--
作者：
Z. He;J. Wiegel
从厌氧生物Clostridium hydroxybenzoicum菌株JW/Z-1 T中分离纯化了一种4-羟基苯甲酸脱羧酶。它的表观分子量为350 kDa，由6个相同的亚基组成，每个亚基为57 kDa。脱羧的最佳温度为约50 ℃，最佳pH为5.6-6.2。该酶的pI为5.1。4-羟基苯甲酸酯脱羧的活化能为65 kJ.mol-1（20-37 ℃）。该酶还催化3，4-二羟基苯甲酸的脱羧。在pH 6.0和25 ℃下，4-羟基苯甲酸酯的表观Km和kcat值分别为0.40 mM和3.3 x 10（3）min-1，3，4-二羟基苯甲酸酯的表观Km和kcat值分别为1.2 mM和1.1 x 10（3）min-1。酶活性不受生物素或抗生物素蛋白添加到粗细胞提取物或纯化的酶。羟基苯甲酸酯的对羟基基团似乎是酶结合所必需的。N-末端氨基酸序列显示出与来自聚球藻属和酵母属的尿卟啉原脱羧酶的一些相似性。该酶催化逆反应，即苯酚羧化为4-羟基苯甲酸和邻苯二酚羧化为3，4-二羟基苯甲酸。羧化不需要ATP。
A 4-hydroxybenzoate decarboxylase from the anaerobe Clostridium hydroxybenzoicum strain JW/Z-1T was purified and partially characterized. It had an apparent molecular mass of 350 kDa and consisted of six identical subunits of 57 kDa each. The temperature optimum for the decarboxylation was approximately 50 degrees C, the optimum pH 5.6-6.2. The pI of the enzyme was 5.1. The activation energy for decarboxylation of 4-hydroxybenzoate was 65 kJ.mol-1 (20-37 degrees C). The enzyme also catalyzed decarboxylation of 3,4-dihydroxybenzoate. The apparent Km and kcat values obtained for 4-hydroxybenzoate were 0.40 mM and 3.3 x 10(3) min-1, and for 3,4-dihydroxybenzoate 1.2 mM and 1.1 x 10(3) min-1, respectively, at pH 6.0 and 25 degrees C. The enzyme activity was not influenced by the addition of biotin or avidin to either the crude cell extracts or the purified enzyme. The p-hydroxyl group of hydroxybenzoate appears to be essential for binding by the enzyme. The N-terminal amino acid sequence shows some similarity to the uroporphyrinogen decarboxylases from Synechococcus and Saccharomyces. The enzyme catalyzed the reverse reactions, that is, the carboxylation of phenol to 4-hydroxybenzoate and of catechol to 3,4-dihydroxybenzoate. The carboxylation did not require ATP.
通过 hem12 突变等位基因的序列分析鉴定影响酵母尿卟啉原脱羧酶活性的氨基酸变化。
DOI：
10.1042/bj2880753
发表时间：
1992
期刊：
The Biochemical journal
影响因子：
--
作者：
Chelstowska,A;Zoladek,T;Garey,J;Kushner,J;Rytka,J;Labbe-Bois,R
通讯作者：
Labbe-Bois,R