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Biochemical Studies on Acetic Acid Bacteria

醋酸菌的生化研究

基本信息

批准号：
62440014
负责人：
TAKIMOTO Koichi
金额：
$ 3.9万
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 (A)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1990
项目状态：
已结题

项目摘要

Cytochrome al is a classic cytochrome that in the 1930s had already been detected in Acetobacter strains and in the 1950s was identified as a terminal oxidase. However, recent studies did not substantiate the previous observations. We have detected a cytochrome al like chromophore in Acetobacter aceti, which was purified and charactarized in this study. The cytochrome was solubilized from membranes of the strain with octyl glucoside and was purified by single column chromatography. The purified cytochrome exhibited a broad alpha peak around 600-610 nm, which turned to a sharp peak at 589 nm in the presence of cyanide. Carbon monoxide difference spectra of the cytochrome indicated the presence of an alphatipe cytochrome. The cytochrome contained l mol each of hemes b and probably one copperion. These results suggest that the cytochrome purified from A。 aceti is the so-called cytochrome al, and thus the existence of the classic cytochrome has been reconfirmed. The purified enzyme consisted of four polypeptides of 55, 35, 22 and 18 kDa, and it showed a sedimentation coefficient of 6.3s in the native form. The enzyme had a high ubiquinol oxidase activity (140-160 micromoles of ubiquinol oxidized per min per mg of protein). When reconstituted into proteoliposomes, the cytochrome could generate an electrochemical proton gradient during oxidation of ubiquinol. Thus, cytochrome al of A。 aceti has been shown to be a cytochrome ba terminal oxidase capable of generating an electrochemical proton gradient concomitant with ubiquinol oxidation.

细胞色素a1是一种经典的细胞色素，在20世纪30年代已在醋杆菌菌株中检测到，并在20世纪50年代被鉴定为末端氧化酶。然而，最近的研究并没有证实之前的观察结果。我们在醋酸醋杆菌中检测到细胞色素 a1 样发色团，并在本研究中对其进行了纯化和表征。用辛基葡萄糖苷从菌株膜上溶解细胞色素，并通过单柱层析纯化。纯化的细胞色素在 600-610 nm 附近表现出宽 α 峰，在氰化物存在下，该峰在 589 nm 处变成尖峰。细胞色素的一氧化碳差异光谱表明αtipe细胞色素的存在。细胞色素含有各 1 mol 血红素 b 和可能一个铜离子。这些结果表明，从A.aceti中纯化的细胞色素就是所谓的细胞色素al，从而再次证实了经典细胞色素的存在。纯化的酶由55、35、22和18 kDa的四种多肽组成，其天然形式的沉降系数为6.3s。该酶具有高泛醇氧化酶活性（每分钟每毫克蛋白质氧化 140-160 微摩尔泛醇）。当重组为蛋白脂质体时，细胞色素可以在泛醇氧化过程中产生电化学质子梯度。因此，细胞色素al of A。 aceti已被证明是一种细胞色素ba末端氧化酶，能够伴随泛醇氧化产生电化学质子梯度。