α-Glycerophosphate dehydrogenase

作者：David Ezra Green · 发表于：Biochemical Journal · 年份：1936 · DOI：10.1042/bj0300629 · 被引用次数：148 · 研究领域：Metabolism and Genetic Disorders、Enzyme Catalysis and Immobilization、Lipid metabolism and biosynthesis

MEYERHOF [1919] was the first to observe the oxidation of glycerophosphate by muscle and liver tissue.Ahlgren [1925], Quastel and Whetham [1925], Quastel and Wooldridge [1927], Alwal [1928; 1929], Colett et al. [1929], Davies and Quastel [1932] and McGavran and Rheinberger [1933] have considered some of the properties of the glycerophosphate dehydrogenase.There has not been however any systematic investigation of this enzyme, and there are no data available as to the method of preparation, the conditions for maximum activity, the nature of the oxidation product or the mechanism of the reaction with molecular oxygen.I. Preparation of the enzyme.The dissected skeletal muscles of a freshly killed rabbit were passed twice through a coarse meat mincer, and washed exhaustively with tap water.The washed mince was mixed with sand and 500 ml.distilled water and ground to a paste in a mechanical mortar.The sand and insoluble debris were filtered off through muslin.50 ml. of M/10 acetate buffer of PH 4-6 were added to the filtrate, and the precipitate was centrifuged.The supernatant fluid was dis- carded and the precipitate was resuspended in 100 ml.M/5 phosphate buffer of pl1 7-2.The enzyme suspension retains the bulk of its activity for a period of 10 days if kept at 00.There is a definite fall in activity even at this low tem- perature.The precipitate can also be dried, in vacuo.The enzyme in the dried