Solubilization, Purification and Properties of Membrane-bound Glycerol Dehydrogenase from Gluconobacter industrius
Solubilization, Purification and Properties of Membrane-bound Glycerol Dehydrogenase from Gluconobacter industrius
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工业葡糖杆菌膜结合甘油脱氢酶的溶解、纯化和性质
DOI：
10.1080/00021369.1985.10866858
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
O. Adachi
中科院分区：
文献类型：
--
作者：
M. Ameyama;E. Shinagawa;K. Matsushita;O. Adachi
膜结合的甘油脱氢酶溶解和纯化约100倍的膜上生长的Gluconetraindustrius IFO 3260的甘油-谷氨酸培养基。通过在0.05 M Tris-HCl（pH 8.0）中使用0.5%二甲基十二烷基氧化胺成功地实现了酶的溶解。乙醇脱氢酶和d-葡萄糖脱氢酶，这是丰富的形成在同一细菌膜，消除增溶。甘油脱氢酶通过用聚乙二醇6000分级进一步纯化。该酶表现出广泛的底物特异性和各种多羟基醇，除了甘油，在2，6-二氯酚靛酚和吩嗪硫酸甲酯作为电子受体的存在下，迅速氧化，但NAD和NADP是惰性的。该酶是一种以吡咯喹啉醌为辅基的醌蛋白。
Membrane-bound glycerol dehydrogenase was solubilized and purified about 100-fold from the membrane of Gluconobacter industrius IFO 3260 grown on a glycerol-glutamate medium. Solubilization of the enzyme was successfully achieved by use of 0.5% dimethyldodecylamineoxide in 0.05 M Tris-HCl, pH 8.0. Alcohol dehydrogenase and d-glucose dehydrogenase, which were abundantly formed in the same bacterial membrane, were eliminated on solubilization. Glycerol dehydrogenase was further purified through fractionation with polyethylene glycol 6000. The enzyme showed a broad substrate specificity and various kinds of polyhydroxyl alcohols, in addition to glycerol, were rapidly oxidized in the presence of 2,6-dichlorophenolindophenol and phenazine methosulfate as the electron acceptor but NAD and NADP were inert. The enzyme was proved to be a quinoprotein in which pyrroloquinoline quinone functioned as the prosthetic group.