恶臭假单胞菌N.C.I.B. 9896中的对甲酚和3,5-二甲苯酚甲基羟化酶
P-cresol and 3,5-xylenol methylhydroxylases in Pseudomonas putida N.C.I.B. 9896.
作者信息
Keat M J, Hopper D J
出版信息
Biochem J. 1978 Nov 1;175(2):649-58. doi: 10.1042/bj1750649.
Abstract
Pseudomonas putida N.C.I.B. 9869, when grown on 3,5-xylenol, hydroxylates the methyl groups on 3,5-xylenol and on p-cresol by two different enzymes. 3,5-Xylenol methylhydroxylase, studied only in relatively crude extracts, requires NADH, is not active with p-cresol and is inhibited by cyanide, but not by CO. The p-cresol methylhydroxylase requires an electron acceptor and will act under anaerobic conditions. It was purified and is a flavocytochrome c of mol.wt. approx. 114,000 consisting of two subunits of equal size. The enzyme catalyses the hydroxylation of p-cresol (Km 16 micron) and the further oxidation of product, p-hydroxybenzyl alcohol (Km 27 micron) to p-hydroxybenzaldehyde. A different p-cresol methylhydroxylase of the flavocytochrome c type is induced by growth on p-cresol. It too was purified and has mol.wt. approx. 100,000, and again consisted of two equal-size subunits. The Km for p=cresol 3.6 micron and for p=hydroxybenzyl alcohol, 15 micron.
摘要
恶臭假单胞菌N.C.I.B. 9869在以3,5-二甲苯酚为底物生长时，通过两种不同的酶将3,5-二甲苯酚和对甲酚上的甲基羟基化。3,5-二甲苯酚甲基羟化酶仅在相对粗提物中进行了研究，它需要NADH，对对甲酚无活性，受氰化物抑制，但不受CO抑制。对甲酚甲基羟化酶需要一个电子受体，并且在厌氧条件下也能起作用。它被纯化出来，是一种分子量约为114,000的黄素细胞色素c，由两个大小相等的亚基组成。该酶催化对甲酚（Km为16微摩尔）的羟基化以及产物对羟基苄醇（Km为27微摩尔）进一步氧化为对羟基苯甲醛。在以对甲酚为底物生长时可诱导产生一种不同的黄素细胞色素c型对甲酚甲基羟化酶。它也被纯化出来，分子量约为100,000，同样由两个大小相等的亚基组成。对甲酚的Km为3.6微摩尔，对羟基苄醇的Km为15微摩尔。
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