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Functionnal Characters of Alcohol and Methanol Dehydrogenases in the Respiratory Chain of Acetic Acid Bacteria

乙酸菌呼吸链中醇和甲醇脱氢酶的功能特征

基本信息

批准号：
02660122
负责人：
MATSUSHITA Kazunobu
金额：
$ 1.28万
依托单位：
Yamaguchi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

Acetobacter methanolicus is a unique acetic acid bacterium which has a methanol oxidase respiratory chain, as seen in methylotrophs, in addition to its ethanol oxidase respiratory chain. In this study, the relationship between methanol and ethanol oxidase respiratory chains was investigated. Furhtermore, in this study, the structure and function of methanol dehydrogenase (MDH) purified from A. methatiolicus and alcohol dehydrogenases (ADH) purified from several acetic acid bacteria including A. metizanolicus. (1) A. methanolicus grown on methanol exhibited a high methanol oxidase activity and contained a cytochrome c oxidase, cytochrome co, while the cells grown on glycerol showed higher ethanol oxidase activity and contained an ubiquinol oxidase, cytochrome bo. The results obtained in this study suggest that A. methanolicus has two independent respiratory chains for methanol and ethanol. The methanol oxidase respiratory chain appears to operate by linking MDH to cytochrome c oxidase, … More cytochrome co, via soluble cytochrome(s) c, while the ethanol oxidase respiratory chain consists of ADH, ubiquinone and cytochrome bo ubiquinol oxidase. (2) Two different types of MDH were purified from A. methanolicus grown on methanol. One had an alpha2gamma2structure while the other did an alpha2gamma2 structure. Both types of enzyme had been detected in the periplasm at a ratio of 7 : 3, respectively. The results obtained here suggested that the alpha2gamma2 MDH may be produced from the alpha2gamma2 enzyme during the preparation. (3) ADH was purified from the membranes of glycerol-grown cells and shown to reduce ubiquinone-10 as well as a short side-chain homologue in detergent solution. The ethanol oxidase respiratory chain was reconstituted into proteoliposomes containing ADH together with ubiquinone-10 and cytochrome bo. Such a reconstitution has been established with ADHs purified from both Acetobacter aceti and Gluconobacter suboxydans. G. suboxydans ADH was dissociated into the subunit I, III and the subunit II, which were shown to be a quinohemoprotein dehydrogenase portion and diheme cytochrome c, respectively. Less

甲醇醋杆菌是一种独特的乙酸细菌，除了其乙醇氧化酶呼吸链之外，还具有甲醇氧化酶呼吸链，如在甲基营养型中所见。本文研究了甲醇和乙醇氧化酶呼吸链之间的关系。本研究还对从A.从几种乙酸细菌中纯化的产甲醇菌和乙醇脱氢酶（ADH），包括A.梅蒂扎诺里库斯(1)A.在甲醇上生长的细胞表现出高的甲醇氧化酶活性，并含有细胞色素C氧化酶，细胞色素CO，而在甘油上生长的细胞表现出更高的乙醇氧化酶活性，并含有泛醇氧化酶，细胞色素B 0。本研究结果表明，A.产甲醇菌对甲醇和乙醇具有两个独立的呼吸链。甲醇氧化酶呼吸链似乎是通过将MDH连接到细胞色素c氧化酶来运作的， ...更多信息乙醇氧化酶呼吸链由ADH、泛醌和细胞色素bo泛醇氧化酶组成。(2)从A.在甲醇上生长的嗜甲醇菌。一个具有α 2 γ 2结构，而另一个具有α 2 γ 2结构。这两种类型的酶已被检测到在周质中的比例为7：3，分别。这里获得的结果表明，α 2 γ 2 MDH可以在制备过程中从α 2 γ 2酶产生。(3)ADH从甘油生长的细胞的膜中纯化，并显示出减少泛醌-10以及洗涤剂溶液中的短侧链同系物。将乙醇氧化酶呼吸链重组到含有ADH、泛醌-10和细胞色素bo的蛋白脂质体中。已经用从醋化醋杆菌和弱氧化葡糖酸杆菌纯化的ADH建立了这样的重构。G. suboxaborans ADH被解离成亚基I、III和亚基II，它们分别被证明是醌血红素蛋白脱氢酶部分和二血红素细胞色素c。少