来自草分枝杆菌ATCC354的β-羟基丁酸脱氢酶的纯化及性质

Purification and properties of beta-hydroxybutyrate dehydrogenase from Mycobacterium phlei ATCC354.

作者信息

Dhariwal K R, Venkitasubramanian T A

出版信息

J Gen Microbiol. 1978 Jan;104(1):123-6. doi: 10.1099/00221287-104-1-123.

Abstract

beta-Hydroxybutyrate dehydrogenase (EC 1.1.1.30) was purified 145-fold from Mycobacterium phlei ATCC354 by ammonium sulphate fractionation and DEAE-cellulose chromatography. The pH optima for oxidation and reduction reactions were 8.4 and 6.8 respectively. The purified enzyme was specific for NAD, NADH, acetoacetate and D(-)-beta-hydroxybutyrate. Km values for DL-beta-hydroxybutyrate and NAD were 7.4 mM and 0.66 mM respectively. The enzyme was inactivated by mercurial thiol inhibitors and by heat, but could be protected by NADH, Ca2+ and partially by Mn2+. The enzyme did not require metal ions and was insensitive to EDTA, glutathione, dithiothreitol, beta-mercaptoethanol and cysteine.

摘要

通过硫酸铵分级分离和DEAE - 纤维素色谱法从草分枝杆菌ATCC354中纯化出β-羟丁酸脱氢酶（EC 1.1.1.30），纯化倍数为145倍。氧化反应和还原反应的最适pH分别为8.4和6.8。纯化后的酶对NAD、NADH、乙酰乙酸和D（-）-β-羟丁酸具有特异性。DL-β-羟丁酸和NAD的Km值分别为7.4 mM和0.66 mM。该酶被汞硫醇抑制剂和热灭活，但可被NADH、Ca2 +保护，部分受Mn2 +保护。该酶不需要金属离子，对EDTA、谷胱甘肽、二硫苏糖醇、β-巯基乙醇和半胱氨酸不敏感。