RECONSTITUTION OF THE ETHANOL OXIDASE RESPIRATORY-CHAIN IN MEMBRANES OF QUINOPROTEIN ALCOHOL DEHYDROGENASE-DEFICIENT GLUCONOBACTER-SUBOXYDANS SUBSP ALPHA-STRAINS
RECONSTITUTION OF THE ETHANOL OXIDASE RESPIRATORY-CHAIN IN MEMBRANES OF QUINOPROTEIN ALCOHOL DEHYDROGENASE-DEFICIENT GLUCONOBACTER-SUBOXYDANS SUBSP ALPHA-STRAINS
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DOI：
10.1128/jb.173.11.3440-3445.1991
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发表时间：
1991-06-01
影响因子：
3.2
通讯作者：
AMEYAMA, M
中科院分区：
文献类型：
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作者：
MATSUSHITA, K;NAGATANI, YI;AMEYAMA, M
利用G.亚黄壳菌亚种alpha，G.不能氧化乙醇的低氧烷。 G.亚黄壳菌亚种α既不表现出醇脱氢酶、乙醇氧化酶，也不表现出葡萄糖-铁氰化物氧化还原酶活性。 此外，与G. suboxidans。 乙醇脱氢酶复合物的第一亚基醌血红素蛋白和第二亚基细胞色素c在G.亚黄壳菌亚种通过使用血红素染色和免疫印迹方法，分别显示α减少和缺乏。 乙醇氧化酶活性，G.亚黄壳菌亚种α，完全恢复重组乙醇脱氢酶纯化G。suboxetans的膜G.亚黄壳菌亚种α;这也导致呼吸链中氰化物或叠氮化物不敏感性和葡萄糖-铁氰化物氧化还原酶活性的恢复，而不影响其它呼吸活性如葡萄糖和山梨醇氧化酶。 乙醇氧化酶活性也重建只有第二亚基细胞色素c的酶复合物。 结果表明，乙醇脱氢酶复合物的第二亚基细胞色素c是乙醇氧化酶呼吸链中必需的，可能参与了G. suboxidans。
The ethanol oxidase respiratory chain of Gluconobacter suboxydans was characterized by using G. suboxydans subsp. alpha, a variant species of G. suboxydans incapable of oxidizing ethanol. The membranes of G. suboxydans subsp. alpha exhibited neither alcohol dehydrogenase, ethanol oxidase, nor glucose-ferricyanide oxidoreductase activity. Furthermore, the respiratory chain of the organism exhibited an extremely diminished amount of cytochrome c and an increased sensitivity of the respiratory activity for cyanide or azide when compared with G. suboxydans. The first-subunit quinohemoprotein and the second-subunit cytochrome c of alcohol dehydrogenase complex in the membranes of G. suboxydans subsp. alpha were shown to be reduced and deficient, respectively, by using heme-staining and immunoblotting methods. Ethanol oxidase activity, lacking in G. suboxydans subsp. alpha, was entirely restored by reconstituting alcohol dehydrogenase purified from G. suboxydans to the membranes of G. suboxydans subsp. alpha; this also led to restoration of the cyanide or azide insensitivity and the glucose-ferricyanide oxidoreductase activity in the respiratory chain without affecting other respiratory activities such as glucose and sorbitol oxidases. Ethanol oxidase activity was also reconstituted with only the second-subunit cytochrome c of the enzyme complex. The results indicate that the second-subunit cytochrome c of the alcohol dehydrogenase complex is essential in ethanol oxidase respiratory chain and may be involved in the cyanide- or azide-insensitive respiratory chain bypass of G. suboxydans.