Kawaguchi A, Hosaka K, Kikuchi T, Soda K, Okuda S
J Biochem. 1983 Aug;94(2):487-92. doi: 10.1093/oxfordjournals.jbchem.a134379.
A method for measuring free fatty acids by enzymic cycling is described. Free fatty acids are converted to acyl-CoAs by acyl-CoA synthetase, then the acyl-CoAs are hydrolyzed back to the free fatty acids by acyl-CoA hydrolase in a cyclic fashion. The amounts of AMP produced during this cyclic reaction are determined from the absorbance at 340 nm in the presence of AMP deaminase and glutamate dehydrogenase. This method is sensitive to as low as 0.1 nmol of free fatty acids, and the standard curve is linear up to 1.0 nmol. This method shows a broad specificity for long-chain fatty acids (C12--C20) and the recoveries of fatty acids added to bacterial cell-free extracts are more than 90%.
描述了一种通过酶循环法测量游离脂肪酸的方法。游离脂肪酸通过酰基辅酶A合成酶转化为酰基辅酶A，然后酰基辅酶A通过酰基辅酶A水解酶以循环方式水解回游离脂肪酸。在存在AMP脱氨酶和谷氨酸脱氢酶的情况下，根据340nm处的吸光度测定该循环反应过程中产生的AMP量。该方法对低至0.1nmol的游离脂肪酸敏感，标准曲线在1.0nmol范围内呈线性。该方法对长链脂肪酸（C12 - C20）具有广泛的特异性，添加到无细胞细菌提取物中的脂肪酸回收率超过90%。
