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来自芳香陶厄氏菌的苯甲酸辅酶A连接酶:一种作用于厌氧和好氧途径的酶。

Benzoate-coenzyme A ligase from Thauera aromatica: an enzyme acting in anaerobic and aerobic pathways.

作者信息

Schühle Karola, Gescher Johannes, Feil Ulrich, Paul Michael, Jahn Martina, Schägger Hermann, Fuchs Georg

机构信息

Mikrobiologie, Institut für Biologie II, Universität Freiburg, Germany.

出版信息

J Bacteriol. 2003 Aug;185(16):4920-9. doi: 10.1128/JB.185.16.4920-4929.2003.

Abstract

In the denitrifying member of the beta-Proteobacteria Thauera aromatica, the anaerobic metabolism of aromatic acids such as benzoate or 2-aminobenzoate is initiated by the formation of the coenzyme A (CoA) thioester, benzoyl-CoA and 2-aminobenzoyl-CoA, respectively. Both aromatic substrates were transformed to the acyl-CoA intermediate by a single CoA ligase (AMP forming) that preferentially acted on benzoate. This benzoate-CoA ligase was purified and characterized as a 57-kDa monomeric protein. Based on V(max)/K(m), the specificity constant for 2-aminobenzoate was 15 times lower than that for benzoate; this may be the reason for the slower growth on 2-aminobenzoate. The benzoate-CoA ligase gene was cloned and sequenced and was found not to be part of the gene cluster encoding the general benzoyl-CoA pathway of anaerobic aromatic metabolism. Rather, it was located in a cluster of genes coding for a novel aerobic benzoate oxidation pathway. In line with this finding, the same CoA ligase was induced during aerobic growth with benzoate. A deletion mutant not only was unable to grow anaerobically on benzoate or 2-aminobenzoate, but also aerobic growth on benzoate was affected. This suggests that benzoate induces a single benzoate-CoA ligase. The product of benzoate activation, benzoyl-CoA, then acts as inducer of separate anaerobic or aerobic pathways of benzoyl-CoA, depending on whether oxygen is lacking or present.

摘要

在β-变形菌纲的反硝化菌陶厄氏菌属中,苯甲酸或2-氨基苯甲酸等芳香酸的厌氧代谢分别通过辅酶A(CoA)硫酯即苯甲酰-CoA和2-氨基苯甲酰-CoA的形成而启动。两种芳香底物都通过一种优先作用于苯甲酸的单一CoA连接酶(形成AMP)转化为酰基-CoA中间体。这种苯甲酸-CoA连接酶被纯化并鉴定为一种57 kDa的单体蛋白。根据V(max)/K(m),2-氨基苯甲酸的特异性常数比苯甲酸低15倍;这可能是在2-氨基苯甲酸上生长较慢的原因。苯甲酸-CoA连接酶基因被克隆并测序,发现它不是编码厌氧芳香代谢通用苯甲酰-CoA途径的基因簇的一部分。相反,它位于编码一条新的需氧苯甲酸氧化途径的基因簇中。与此发现一致的是,在用苯甲酸进行需氧生长期间诱导出了相同的CoA连接酶。一个缺失突变体不仅不能在苯甲酸或2-氨基苯甲酸上厌氧生长,而且在苯甲酸上的需氧生长也受到影响。这表明苯甲酸诱导产生一种单一的苯甲酸-CoA连接酶。苯甲酸激活的产物苯甲酰-CoA随后根据是否缺氧分别作为苯甲酰-CoA不同厌氧或需氧途径的诱导剂。

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