改进铜皂-分光光度法测定脂肪酶活力  

A Modified Colorimetric Assay of Lipase activity Using Emulsified Olive Oil as the Substrate

作　　者：侯爱军[1] 徐冰斌[1] 梁亮[1] 李艳红[1] 彭必雨[1,2] 

机构地区：[1]四川大学轻纺与食品学院,四川成都610065 [2]四川大学制革清洁技术国家工程实验室,四川成都610065

出　　处：《皮革科学与工程》2011年第1期22-27,共6页Leather Science and Engineering

基　　金：国家科技支撑计划项目(2006BAC02A09)

摘　　要：建立了一种改进的脂肪酶活力的检测方法。以乳化橄榄油为底物,用乙醇和盐酸终止酶反应,并使底物乳液破乳,用异辛烷萃取生成的脂肪酸,并用醋酸铜显色,通过铜皂的吸光度来测定生成的脂肪酸的量而得到脂肪酶的活力。主要研究了异辛烷萃取乳化体系中脂肪酸的条件,测定了脂肪酸的萃取百分率和该方法检测的线性范围。用改进的方法对两种脂肪酶的活力进行了检测,并与传统的滴定法进行了对比。结果表明,在优化的萃取条件下脂肪酸的回收率为83.68%,该方法的线性范围为吸光度(OD值)0.154～0.291。改进的铜皂-分光光度法测定结果的重复性优于滴定法。由于该方法的测定不受反应体系的缓冲性的影响,测得的酶活力-pH曲线更准确。A modified colorimetric method for assaying lipase activity was developed.Lipase hydrolyzes emulsified olive oil,and produced fatty acid was extracted by isooctane instead of benzene,then the extracted fatty acid was determined by copper soap colorimetry measurement.The effective method for extracting free fatty acids in emulsion with isooctane and the extraction ratio were investigated.The results show that the recovery of fatty acid is 83.68%,and the linear OD range is 0.154-0.291.The reproducibility of the modified method is much better than that of titration method.The curves of lipase activities with pHs are more accurate from the modified colorimetric assay.

关 键 词：脂肪酶活力测定 分光光度法 乳化橄榄油 脂肪酸 

分 类 号：TS57[轻工技术与工程—皮革化学与工程]