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Author:
Date:
2012-09-05
[Abstract] Acetyl-coenzyme A synthethase (Acs, E.C.6.2.1.1) is an acetate activating enzyme widely represented in nature from bacteria to human. Its function is important for cellular catabolism, especially in order to support microbial growth at low concentrations of acetate (<10 mm)="" (castano="" cerezo="">et al., 2011; Castano Cerezo et al., 2009；Renilla et al., 2012). In this protocol, a continuous coupled enzymatic assay for Acs activity is described. Product formation is followed spectrophotometrically by the formation of NADH. The protocol is tailored for E. coli’s Acs, but it can be adapted to assay Acs in any other organism.
The acetyl-coenzyme A synthetase (Acs) assay was first described by Brown et al. (1977). Acs activity is ...
[摘要] 乙酰辅酶A合成酶（Acs，E.C.6.2.1.1）是从细菌到人广泛代表的乙酸活化酶。其功能对于细胞分解代谢是重要的，特别是为了支持在低浓度的乙酸盐（<10mm）下的微生物生长（castano cerezo等人，2011;="" castano="" cerezo等人，="" ，2009;="">
首先由Brown等人描述了乙酰辅酶A合成酶（Acs）测定法 。 （1977）。使用与苹果酸脱氢酶（Mdh）和柠檬酸合酶（Cs）偶联的酶促方法测量Acs活性：乙酸+ AcASH + ATP→乙酰-CoA + AMP（Cs）乙酰 - CoA +草酰乙酸盐→柠檬酸盐+ Co Wash
（Mdh）L-苹果酸盐+ NAD +→草酰乙酸盐+ NADH
净反应：乙酸盐+ ATP + L-苹果酸盐+ NAD +→柠檬酸盐+ AMP + NADH />在测定条件下，Mdh和Cs活性过量，NADH形成速率受Acs活性限制。
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