214型离子交换树脂固定化假丝酵母脂肪酶的研究

摘要

以假丝酵母脂肪酶(Candida rugosa lipase)催化脂肪酸甲酯化是植物油精制副产物中提取天然维生素E的重要预处理反应.为提高预处理效果,通过固定化假丝酵母脂肪酶提高其酯化能力,降低水解反应的能力.笔者在不同性能树脂筛选的基础上,开展了离子交换树脂214型固定化假丝酵母脂肪酶的研究.以酶固定化率为指标,开展了酶质量浓度、树脂的量、缓冲液pH、固定化温度、时间和振荡速率等条件试验和响应面试验,得到的最适固定化条件为酶质量浓度6.09 mg/mL,5 mL酶液中加入0.75 g树脂,缓冲液pH 8.4,振荡转速134.26 r/min,固定化温度30℃,固定化时间4h.该条件下,假丝酵母脂肪酶与树脂的结合率最高,酶的水解作用基本消失,酯化能力得到保持,可以获得较好的维生素E提纯预处理效果.%Fatty acid catalytic esterification by Candida rugosa lipase is an important preconditioning response in the extraction of natural Vitamin E from by-products of vegetable oil refining.In order to improve the efficiency of pretreatment,immobilized lipase was applied to improve the esterification capacity and to reduce the hydrolysis reaction.214 ion exchange resin was selected to immobilize Candida rugosa lipase,the concentration of enzyme,the amount of the resin,buffer solution pH,temperature,time and oscillation rate condition tests and response surface test were made while the reaction rate of the immobilized enzymes was taken as an index.The optimum conditions were:30℃,0.75 g resin,6.09 mg/mL enzyme,pH 8.40,oscillation speed 134.26 r/min,4 h.Under this condition,the hydrolysis reaction was disappeared,immobilization rate reached a maximum.