PURIFICATION AND CHARACTERIZATION OF PARTICULATE ALCOHOL-DEHYDROGENASE FROM GLUCONOBACTER-SUBOXYDANS
PURIFICATION AND CHARACTERIZATION OF PARTICULATE ALCOHOL-DEHYDROGENASE FROM GLUCONOBACTER-SUBOXYDANS
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DOI：
10.1080/00021369.1978.10863306
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发表时间：
1978-01-01
期刊：
影响因子：
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通讯作者：
AMEYAMA, M
中科院分区：
文献类型：
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作者：
ADACHI, O;TAYAMA, K;AMEYAMA, M
从醋酸菌G. [oxydans ssp.] subboxerans IFO 12528.用TritonX-100溶解菌体膜组分，然后用DEAE-SephadexA-50和羟基磷灰石分级分离，成功地纯化了酶。细胞色素c样组分与脱氢酶蛋白紧密结合，并以酶-细胞色素复合物的形式存在。醇脱氢酶本身不是细胞色素组分。该酶的分子量为150，000，凝胶电泳显示存在分子量为85，000、49，000和14，400的三个亚基。最小的亚基对应于细胞色素c样组分。乙醇在体外染料存在下被氧化，但不需要NAD或NADP作为H受体。与酵母或[哺乳动物]肝脏中的NAD-连接的醇脱氢酶和甲醇利用细菌中的其他初级醇脱氢酶不同，来自乙酸细菌的酶在相当酸性的pH下显示其最适pH。
Particulate alcohol dehydrogenase of acetic acid bacteria that is mainly participated in vinegar fermentation was purified to homogeneous state from G. [oxydans ssp.] suboxydans IFO 12528. Solubilization of enzyme from the bacterial membrane fraction by Triton X-100 and subsequent fractionation on DEAE-Sephadex A-50 and hydroxylapatite was successful in enzyme purification. A cytochrome c-like component was tightly bound to the dehydrogenase protein and existed as an enzyme-cytochrome complex. The alcohol dehydrogenase is not a cytochrome component itself. The MW of the enzyme was 150,000, and gel electrophoresis showed the presence of three subunits having a MW of 85,000, 49,000 and 14,400. The smallest subunit was corresponded to the cytochrome c-like component. Ethanol was oxidized in the presence of dyes in vitro but NAD or NADP were not required as H acceptor. Unlike NAD-linked alcohol dehydrogenase in yeast or [mammalian] liver and other primary alcohol dehydrogenases in methanol utilizing bacteria, the enzyme from the acetic acid bacteria showed its optimum pH at fairly acidic pH.