Fell D A, Liddle P F, Peacocke A R, Dwek R A
Biochem J. 1974 Jun;139(3):665-75. doi: 10.1042/bj1390665.
A new method is described for the preparation of pyruvate kinase from yeast. This eliminates proteolysis during the preparation. The molecular weight of yeast pyruvate kinase is 215000, and it is composed of four subunits. Such properties of the enzyme as its extinction coefficient, cold-lability, thiol-group reactivity and binding of Mn(2+) ions are compared with those previously reported for yeast pyruvate kinase prepared by different methods. The specific activity is significantly higher than previously observed, but otherwise the enzyme is similar, apart from its molecular weight and Mn(2+)-binding characteristics, to preparations from Saccharomyces cerevisiae obtained in this laboratory (e.g. Fell et al., 1972, and references therein) and that of C. H. Suelter (e.g. Kuczenski & Suelter, 1971, and references therein), and is different from the enzyme isolated from Saccharomyces carlsbergensis by B. Hess and his co-workers (e.g. Wieker & Hess, 1972, and references therein).
本文描述了一种从酵母中制备丙酮酸激酶的新方法。该方法可避免制备过程中的蛋白水解。酵母丙酮酸激酶的分子量为215000,由四个亚基组成。将该酶的消光系数、冷不稳定性、巯基反应性和锰离子结合等性质与先前报道的用不同方法制备的酵母丙酮酸激酶的性质进行了比较。其比活性显著高于先前观察到的,但除此之外,该酶与本实验室从酿酒酵母中获得的制剂(例如Fell等人,1972年及其中的参考文献)以及C.H.苏尔特的制剂(例如Kuczenski和Suelter,1971年及其中的参考文献)相似,只是分子量和锰离子结合特性有所不同,并且与B.赫斯及其同事从卡尔斯伯酵母中分离出的酶不同(例如Wieker和赫斯,1972年及其中的参考文献)。