PURIFICATION AND CHARACTERIZATION OF OXALYL-COENZYME-A DECARBOXYLASE FROM OXALOBACTER-FORMIGENES
PURIFICATION AND CHARACTERIZATION OF OXALYL-COENZYME-A DECARBOXYLASE FROM OXALOBACTER-FORMIGENES
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DOI：
10.1128/jb.171.5.2605-2608.1989
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发表时间：
1989-05-01
影响因子：
3.2
通讯作者：
ALLISON, MJ
中科院分区：
文献类型：
--
作者：
BAETZ, AL;ALLISON, MJ
通过高压液相色谱结合疏水作用色谱、DEAE 阴离子交换色谱和凝胶渗透色谱从产草酸杆菌中纯化草酰辅酶 A (草酰-CoA) 脱羧酶。该酶由四个相同的亚基 (Mr, 65,000) 组成，从而产生活性酶 (Mr, 260,000)。该酶催化草酰辅酶 A 的硫胺素 PPi 依赖性脱羧反应生成甲酸和二氧化碳。草酰辅酶A的表观Km和V Max 值分别为0.24 mM和0.25 μmol/min，焦磷酸硫胺素的表观Km和V Max 值分别为1.1 pM和0.14 μmol/min。最大比活性为每分钟每毫克蛋白质脱羧 13.5 μM 草酰辅酶A。
Oxalyl-coenzyme A (oxalyl-CoA) decarboxylase was purified from Oxalobacter formigenes by high-pressure liquid chromatography with hydrophobic interaction chromatography, DEAE anion-exchange chromatography, and gel permeation chromatography. The enzyme is made up of four identical subunits (Mr, 65,000) to give the active enzyme (Mr, 260,000). The enzyme catalyzed the thiamine PPi-dependent decarboxylation of oxalyl-CoA to formate and carbon dioxide. Apparent Km and V Max values, respectively, were 0.24 mM and 0.25 .mu.mol/min for oxalyl-CoA and 1.1 pM and 0.14 .mu.mol.min for thiamine pyrophosphate. The maximum specific activity was 13.5 .mu.M oxalyl-CoA decarboxylated per min per mg of protein.