Li H Q, Matsuda I, Fujise Y, Ichiyama A
First Department of Biochemistry Kannondai, Tsukuba, Ibaraki, 305-8604, Japan.
J Biochem. 1999 Jul;126(1):243-53. doi: 10.1093/oxfordjournals.jbchem.a022429.
In Burkholderia glumae (formerly named Pseudomonas glumae), isolated as the causal agent of grain rot and seedling rot of rice, oxalate was produced from oxaloacetate in the presence of short-chain acyl-CoA such as acetyl-CoA and propionyl-CoA. Upon purification, the enzyme responsible was separated into two fractions (tentatively named fractions II and III), both of which were required for the acyl-CoA-dependent production of oxalate. In conjugation with the oxalate production from oxaloacetate catalyzed by fractions II and III, acetyl-CoA used as the acyl-CoA substrate was consumed and equivalent amounts of CoASH and acetoacetate were formed. The isotope incorporation pattern indicated that the two carbon atoms of oxalate are both derived from oxaloacetate, and among the four carbon atoms of acetoacetate two are from oxaloacetate and two from acetyl-CoA. When the reaction was carried out with fraction II alone, a decrease in acetyl-CoA and an equivalent level of net utilization of oxaloacetate were observed without appreciable formation of CoASH, acetoacetate or oxalate. It appears that in the oxalate production from oxaloacetate and acetyl-CoA, fraction II catalyzes condensation of the two substrates to form an intermediate which is split into oxalate and acetoacetate by fraction III being accompanied by the release of CoASH.
在从水稻谷粒腐烂和幼苗腐烂病中分离出的病原菌——伯克霍尔德氏菌(以前称为稻谷假单胞菌)中,草酰乙酸在短链酰基辅酶A(如乙酰辅酶A和丙酰辅酶A)存在的情况下会生成草酸盐。纯化后,负责该反应的酶被分离成两个组分(暂命名为组分II和组分III),这两个组分都是酰基辅酶A依赖性草酸盐生成所必需的。在组分II和组分III催化草酰乙酸生成草酸盐的过程中,用作酰基辅酶A底物的乙酰辅酶A被消耗,并形成等量的辅酶A和乙酰乙酸。同位素掺入模式表明,草酸盐的两个碳原子均来自草酰乙酸,而乙酰乙酸的四个碳原子中,两个来自草酰乙酸,两个来自乙酰辅酶A。当仅用组分II进行反应时,观察到乙酰辅酶A减少,草酰乙酸有相当水平的净利用,但没有明显生成辅酶A、乙酰乙酸或草酸盐。似乎在草酰乙酸和乙酰辅酶A生成草酸盐的过程中,组分II催化两种底物缩合形成一种中间体,该中间体被组分III分解为草酸盐和乙酰乙酸,并伴有辅酶A的释放。