从明胶红假单胞菌中共同纯化柠檬酸裂解酶和柠檬酸裂解酶连接酶，并随后分离这两种酶。

Copurification of citrate lyase and citrate lyase ligase from Rhodopseudomonas gelatinosa and subsequent separation of the two enzymes.

作者信息

Antranikian G, Gottschalk G

出版信息

Eur J Biochem. 1982 Aug;126(1):43-7. doi: 10.1111/j.1432-1033.1982.tb06743.x.

DOI:10.1111/j.1432-1033.1982.tb06743.x

Abstract

A procedure has been worked out which allowed the purification and crystallization of a citrate lyase/citrate lyase ligase complex from Rhodopseudomonas gelatinosa. The complex was subsequently separated to yield two homogeneous enzymes. Citrate lyase ligase was purified 365-fold with a yield of 3.23%. The molecular weight of the enzyme was estimated to be 39500, the enzyme consisted of one polypeptide chain. The reaction rates for ATP, acetate and citrate lyase (sulfhydryl form) followed Michaelis-Menten kinetics (Km values: 0.14 mM, 5 mM and 37 nM respectively). Citrate lyase ligase exhibited a high substrate specificity and could not react with citrate lyases from nonphototrophic microorganisms. In contrast to the ligase from Streptococcus diacetilactis, the enzyme from R. gelatinosa was extremely labile; however, it could be stabilized by nucleotides, the most potent stabilizing one being ADP.

摘要

已经制定出一种方法，可从明胶红假单胞菌中纯化并结晶柠檬酸裂解酶/柠檬酸裂解酶连接酶复合物。随后将该复合物分离以产生两种均一的酶。柠檬酸裂解酶连接酶纯化了365倍，产率为3.23％。该酶的分子量估计为39500，由一条多肽链组成。ATP、乙酸盐和柠檬酸裂解酶（巯基形式）的反应速率遵循米氏动力学（Km值分别为0.14 mM、5 mM和37 nM）。柠檬酸裂解酶连接酶表现出高底物特异性，并且不能与非光合微生物的柠檬酸裂解酶反应。与双乙酰乳链球菌的连接酶相反，明胶红假单胞菌的酶极其不稳定；然而，它可以被核苷酸稳定，最有效的稳定剂是ADP。