Sokolov A P, Luchin S V, Trotsenko Iu A
Biokhimiia. 1980 Aug;45(8):1371-8.
The glucose-6-phosphate and 6-phosphogluconate dehydrogenases i. e. enzymes of dissimilatory hexulose phosphate cycle, were isolated from the cells of the facultative methylotrophic bacterium Pseudomonas oleovorans. The purification procedure included protein fractionation by ammonium sulfate, gel-filtration through Sephacryl S-200 and chromatography on DEAE Bio-Gel A and phosphocellulose, resulting in a 400-fold purification of the enzyme. During analytical disc-electrophoresis in polyacrylamide gel the glucose-6-phosphate dehydrogenase preparation produced a single band; 6-phosphogluconate dehydrogenase was found to contain small contaminations. The molecular weights of the dehydrogenases are 100000 and 110000, respectively. Both enzymes have two subunits. The Km values for glucose-6-phosphate dehydrogenase are 65 mkM for glucose-6-phosphate and 28 mkM for NADP; those for 6-phosphogluconate dehydrogenase are 152 mkM for 6-phosphogluconate and 55 mkM for NADP. Nucleotides are found to be the most active inhibitors of glucose-6-phosphate dehydrogenase. 6-Phosphogluconate dehydrogenase is inhibited by ribose-5-phosphate and fructose-1,6-diphosphate.
从兼性甲基营养细菌食油假单胞菌（Pseudomonas oleovorans）的细胞中分离出了6-磷酸葡萄糖脱氢酶和6-磷酸葡萄糖酸脱氢酶，即异化磷酸己酮糖途径的酶。纯化过程包括用硫酸铵进行蛋白质分级分离、通过Sephacryl S - 200进行凝胶过滤以及在DEAE Bio - Gel A和磷酸纤维素上进行层析，从而使该酶得到了400倍的纯化。在聚丙烯酰胺凝胶的分析圆盘电泳过程中，6-磷酸葡萄糖脱氢酶制剂产生了一条单一的条带；发现6-磷酸葡萄糖酸脱氢酶含有少量杂质。这两种脱氢酶的分子量分别为100000和110000。两种酶都有两个亚基。6-磷酸葡萄糖脱氢酶对6-磷酸葡萄糖的Km值为65μM，对NADP的Km值为28μM；6-磷酸葡萄糖酸脱氢酶对6-磷酸葡萄糖酸的Km值为152μM，对NADP的Km值为55μM。发现核苷酸是6-磷酸葡萄糖脱氢酶最有效的抑制剂。6-磷酸葡萄糖酸脱氢酶受到5-磷酸核糖和1,6-二磷酸果糖的抑制。
