脂肪酸修饰活性蛋白质的工艺研究

目的以牛血清白蛋白为例建立用脂肪酸修饰活性蛋白质的工艺。方法通过脂肪酸的酰氯在酸催化下50℃与N-羟基琥珀酰亚胺反应制备活化酯,再与蛋白质发生偶联形成产物,利用红外谱图对反应产物进行鉴定。结果月桂酸、棕榈酸等脂肪酸与氯化亚砜回馏3 h很容易得到相应的脂肪酰氯,酰氯在酸催化下50℃与N-羟基琥珀酰亚胺反应3 h可以将N-羟基琥珀酰亚胺转变成活性酯,产率为90%,脂肪酸的活性酯在pH=8的缓冲液可以偶联到活性蛋白质上,活化酯转化率为87.9%。结论建立了脂肪酸修饰活性蛋白质的工艺。

A process study on modifying an active protein with fatty acid

Purpose To establish a process to modify an active protein(bovine serum albumin as an example) with Fatty acid.MethodsFatty acid such as lauric acid or palm acid was reacted with thionyl chloride to change into acyl chloride,then acyl chloride interacted with N-hydroxysuccinimide by benzic sulfinic acid catalysis,forming an actived ester,which was coupled with an active protein(BSA as an example)in a PBS buffer(pH=8).The modified protein coupled fatty acid was purified by preparing with cooled acetone and all the intermediate and modified protein was analyzed by IR spectrum.ResultsBy an acid catalysis,the active ester of fatty acid was prepared with 90%yield.IR spectrum conformed that it had a characteristic peak of ester.After being modified by active ester,the protein coupled with fatty acid had characteristics of fatty acid IR.87.9%active ester coupled to protein.ConclusionIt was efficient process to modify active protein by an active ester of fatty acid.