拜氏固氮菌乙酰乙酰辅酶A还原酶的纯化与特性分析
The purification and characterization of acetoacetyl-coenzyme A reductase from Azotobacter beijerinckii.
作者信息
Ritchie G A, Senior P J, Dawes E A
出版信息
Biochem J. 1971 Jan;121(2):309-16. doi: 10.1042/bj1210309.
Abstract
A soluble acetoacetyl-CoA reductase (EC 1.1.1.36) was purified 54-fold from Azotobacter beijerinckii N.C.I.B. 9067 and the reaction product identified as d(-)-beta-hydroxybutyryl-CoA. The Michaelis constants for acetoacetyl-CoA, NADPH and NADH were determined and the reaction rate was found to be some fivefold greater with NADPH than with NADH. At neutral pH the equilibrium greatly favours the formation of the reduced product. Substrate specificity was in the order: acetoacetyl-CoA>acetoacetylpantetheine>acetoacetyl-(acyl-carrier protein). The enzyme possesses a functional thiol group, suffers inactivation by oxygen and is inhibited by thiol-blocking reagents. Inhibition by p-chloromercuribenzoate is reversed by excess of dithiothreitol, which also protects the enzyme from inactivation by oxygen.
摘要
从拜氏固氮菌N.C.I.B. 9067中纯化出一种可溶性乙酰乙酰辅酶A还原酶（EC 1.1.1.36），纯化倍数为54倍，反应产物鉴定为d(-)-β-羟基丁酰辅酶A。测定了该酶对乙酰乙酰辅酶A、NADPH和NADH的米氏常数，发现其对NADPH的反应速率比对NADH的反应速率大约高五倍。在中性pH条件下，平衡极大地有利于还原产物的形成。底物特异性顺序为：乙酰乙酰辅酶A＞乙酰乙酰泛酰巯基乙胺＞乙酰乙酰-（酰基载体蛋白）。该酶具有一个功能性巯基，会被氧气灭活，并受到巯基阻断试剂的抑制。对氯汞苯甲酸的抑制作用可被过量的二硫苏糖醇逆转，二硫苏糖醇还能保护该酶不被氧气灭活。
相似文献
1
The purification and characterization of acetoacetyl-coenzyme A reductase from Azotobacter beijerinckii.拜氏固氮菌乙酰乙酰辅酶A还原酶的纯化与特性分析
Biochem J. 1971 Jan;121(2):309-16. doi: 10.1042/bj1210309.
2
The regulation of poly-beta-hydroxybutyrate metabolism in Azotobacter beijerinckii.拜氏固氮菌中聚-β-羟基丁酸酯代谢的调控
Biochem J. 1973 May;134(1):225-38. doi: 10.1042/bj1340225.
3
An NAD-linked acetoacetyl-CoA reductase from Zoogloea ramigera I-16-M.来自生枝动胶菌I-16-M的一种NAD连接的乙酰乙酰辅酶A还原酶。
Eur J Biochem. 1981 Aug;118(1):53-9. doi: 10.1111/j.1432-1033.1981.tb05485.x.
4
An NADP-linked acetoacetyl CoA reductase from Zoogloea ramigera.来自生枝动胶菌的一种与NADP相关的乙酰乙酰辅酶A还原酶。
Arch Microbiol. 1977 Sep 28;114(3):211-7. doi: 10.1007/BF00446864.
5
Rat hepatic microsomal acetoacetyl-CoA reductase. A beta-ketoacyl-CoA reductase distinct from the long chain beta-ketoacyl-CoA reductase component of the microsomal fatty acid chain elongation system.大鼠肝微粒体乙酰乙酰辅酶A还原酶。一种与微粒体脂肪酸链延长系统的长链β-酮酰基辅酶A还原酶成分不同的β-酮酰基辅酶A还原酶。
J Biol Chem. 1984 Jun 25;259(12):7460-7.
6
Purification and properties of 3-hydroxybutyryl-coenzyme A dehydrogenase from Clostridium beijerinckii ("Clostridium butylicum") NRRL B593.拜氏梭菌（“丁酸梭菌”）NRRL B593中3-羟基丁酰辅酶A脱氢酶的纯化及性质
Appl Environ Microbiol. 1992 Oct;58(10):3297-302. doi: 10.1128/aem.58.10.3297-3302.1992.
7
Acetoacetyl-CoA reductase activity of lactating bovine mammary fatty acid synthase.
J Biol Chem. 1981 Jun 25;256(12):6282-90.
8
Purification and characterization of NADP-linked acetoacetyl-CoA reductase from Zoogloea ramigera I-16-M.来自生枝动胶菌I-16-M的NADP连接的乙酰乙酰辅酶A还原酶的纯化及特性分析
Biochim Biophys Acta. 1987 Feb 23;917(3):365-71.
9
Elementary steps in the reaction mechanism of chicken liver fatty acid synthase: beta-ketoacyl reductase and enoyl reductase.鸡肝脂肪酸合酶反应机制中的基本步骤：β-酮酰基还原酶和烯酰基还原酶。
Biochemistry. 1985 Jan 15;24(2):290-7. doi: 10.1021/bi00323a008.
10
Purification and partial characterization of a methylglyoxal reductase from goat liver.山羊肝脏中甲基乙二醛还原酶的纯化及部分特性分析
Biochim Biophys Acta. 1984 Nov 6;802(1):119-27. doi: 10.1016/0304-4165(84)90041-2.
引用本文的文献
1
Metabolic Engineering Strategies for Enhanced Polyhydroxyalkanoate (PHA) Production in .用于提高……中聚羟基脂肪酸酯（PHA）产量的代谢工程策略
Polymers (Basel). 2025 Jul 31;17(15):2104. doi: 10.3390/polym17152104.
2
Engineering an acetoacetyl-CoA reductase from Cupriavidus necator toward NADH preference under physiological conditions.在生理条件下，对醋酰辅酶 A 还原酶进行工程改造，使其偏向 NADH。
Sci Rep. 2022 Mar 8;12(1):3757. doi: 10.1038/s41598-022-07663-w.
3
Revealing the Metabolic Flexibility of " Accumulibacter phosphatis" through Redox Cofactor Analysis and Metabolic Network Modeling.揭示“聚磷菌”的代谢灵活性通过氧化还原辅因子分析和代谢网络建模。
Appl Environ Microbiol. 2020 Nov 24;86(24). doi: 10.1128/AEM.00808-20.
4
Localization of β-oxidation enzymes in peroxisomes isolated from nonfatty plant tissues.从非脂肪植物组织中分离的过氧化物酶体中β-氧化酶的定位。
Planta. 1983 Nov;159(3):238-46. doi: 10.1007/BF00397531.
5
Control of Enzyme Activities in Cotton Cotyledons during Maturation and Germination : IV. beta-OXIDATION.棉子胚乳中酶活性的控制：IV.β-氧化。
Plant Physiol. 1981 Feb;67(2):341-6. doi: 10.1104/pp.67.2.341.
6
AniA regulates reserve polymer accumulation and global protein expression in Rhizobium etli.AniA调节菜豆根瘤菌中储备聚合物的积累和整体蛋白质表达。
J Bacteriol. 2002 Apr;184(8):2287-95. doi: 10.1128/JB.184.8.2287-2295.2002.
7
Connection between poly-beta-hydroxybutyrate biosynthesis and growth on C(1) and C(2) compounds in the methylotroph Methylobacterium extorquens AM1.甲基营养型细菌嗜甲基嗜酸甲基杆菌AM1中聚-β-羟基丁酸生物合成与利用C(1)和C(2)化合物生长之间的联系。
J Bacteriol. 2001 Feb;183(3):1038-46. doi: 10.1128/JB.183.3.1038-1046.2001.
8
Cloning, molecular analysis, and expression of the polyhydroxyalkanoic acid synthase (phaC) gene from Chromobacterium violaceum.紫色杆菌聚羟基脂肪酸合酶（phaC）基因的克隆、分子分析及表达
Appl Environ Microbiol. 1999 Aug;65(8):3561-5. doi: 10.1128/AEM.65.8.3561-3565.1999.
9
Acetoacetyl coenzyme A reductase and polyhydroxybutyrate synthesis in rhizobium (Cicer) sp. Strain CC 1192.鹰嘴豆根瘤菌（Cicer）菌株CC 1192中的乙酰乙酰辅酶A还原酶与聚羟基丁酸酯合成
Appl Environ Microbiol. 1998 Aug;64(8):2859-63. doi: 10.1128/AEM.64.8.2859-2863.1998.
10
The regulation of poly-beta-hydroxybutyrate metabolism in Azotobacter beijerinckii.拜氏固氮菌中聚-β-羟基丁酸酯代谢的调控
Biochem J. 1973 May;134(1):225-38. doi: 10.1042/bj1340225.
本文引用的文献
1
Colloidal dispersion of chloroplast material.叶绿体物质的胶体分散体。
Science. 1950 Jun 9;111(2893):633-34. doi: 10.1126/science.111.2893.633.
2
Protein measurement with the Folin phenol reagent.使用福林酚试剂进行蛋白质测定。
J Biol Chem. 1951 Nov;193(1):265-75.
3
[SYNTHESIS OF POLY-BETA-HYDROXYBUTYRIC ACID BY HYDROGENOMONAS H-16: PREPARATION OF BETA-HYDROXYBUTYRYL-COENZYME A].[嗜氢菌H-16合成聚-β-羟基丁酸：β-羟基丁酰辅酶A的制备]
Arch Mikrobiol. 1964 Oct 2;49:236-55.
4
ACYL-CARRIER PROTEIN. II. INTERMEDIARY REACTIONS OF FATTY ACID SYNTHESIS.酰基载体蛋白。二、脂肪酸合成的中间反应。
Biochemistry. 1964 Oct;3:1563-71. doi: 10.1021/bi00898a030.
5
Enzymic determination of D(-)-beta-hydroxybutyric acid and acetoacetic acid in blood.血液中D(-)-β-羟基丁酸和乙酰乙酸的酶法测定
Biochem J. 1962 Jan;82(1):90-6. doi: 10.1042/bj0820090.
6
7
Some enzymic reactions concerned in the metabolism of acetoacetyl-coenzyme A in Athiorhodaceae.与紫硫细菌中乙酰乙酰辅酶A代谢相关的一些酶促反应。
Biochem J. 1961 Jul;80(1):70-7. doi: 10.1042/bj0800070.
8
Enzymes of fatty acid metabolism. I. General introduction; crystalline crotonase.脂肪酸代谢的酶。I. 概述；结晶巴豆酸酶
J Biol Chem. 1956 Feb;218(2):971-83.
9
A colorimetric method for the estimation of acetoacetate.一种用于估算乙酰乙酸的比色法。
Biochem J. 1954 Dec;58(4):699-704. doi: 10.1042/bj0580699.
10
Studies on the fatty acid oxidizing system of animal tissues. II. beta-Ketoacyl derivatives of coenzyme A.动物组织脂肪酸氧化系统的研究。II. 辅酶A的β-酮酰基衍生物。
J Biol Chem. 1953 Dec;205(2):575-84.
Zotero 插件