一种用于研究酵母脂肪酸合酶机制的新型发色底物的开发与应用。
The development and application of a novel chromophoric substrate for investigation of the mechanism of yeast fatty acid synthase.
作者信息
Singh N, Wakil S J, Stoops J K
出版信息
Biochem Biophys Res Commun. 1985 Sep 16;131(2):786-92. doi: 10.1016/0006-291x(85)91308-7.
The acetyl transacylase activity of the fatty acid synthase from yeast has been investigated using p-nitrophenylthiol acetate. The chromophoric nature of the nitrophenylthiol moiety affords a convenient spectrophotometric assay for the transacylase function as well as a means to investigate the kinetics and the mechanism of this process. A probable kinetic scheme for enzyme catalyzed transacetylation from p-nitrophenylthiol acetate to an acyl acceptor (CoA or N-acetylcysteamine) is proposed and the kinetic constants for acetylation of enzyme and for acetyl transfer to an acceptor were determined. It was also demonstrated that p-nitrophenylthiol acetate can replace acetyl-CoA as a substrate in fatty acid synthesis.
利用对硝基苯硫醇乙酸酯对酵母脂肪酸合酶的乙酰基转移酶活性进行了研究。对硝基苯硫醇部分的发色性质为转酰基酶功能提供了一种便捷的分光光度测定法，同时也是研究该过程动力学和机制的一种手段。提出了酶催化对硝基苯硫醇乙酸酯与酰基受体（辅酶A或N - 乙酰半胱氨酸）之间转乙酰化的可能动力学方案，并测定了酶乙酰化以及乙酰基转移至受体的动力学常数。还证明了对硝基苯硫醇乙酸酯可替代乙酰辅酶A作为脂肪酸合成的底物。
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