固定化生物印迹假丝酵母脂肪酶的制备研究  

Preparation of Immobilized Bio-Imprinted Candida Rugosa Lipase

作　　者：俞明[1] 钱俊青[1] 龚峰[1] 唐弋奥 YU Ming;QIAN Jun-qing;GONG Feng;TANG Yi-ao(College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, China)

机构地区：[1]浙江工业大学药学院,浙江杭州310014

出　　处：《高校化学工程学报》2017年第4期991-995,共5页Journal of Chemical Engineering of Chinese Universities

基　　金：国家重点研发计划课题(2016YFD0400803)

摘　　要：将生物印迹和固定化酶方法结合,简化了印迹酶的制备流程,优化了固定化印迹脂肪酶的制备条件,筛选了最优印迹分子橄榄油,探讨了印迹最适pH值、印迹分子橄榄油的用量、印迹时间和载体离子型树脂的种类等影响因素对固定化印迹脂肪酶酯化效果的影响,得到了其制备的最佳工艺:印迹pH值8.0,橄榄油100 mg,1 mL乙醇为助溶剂,100 mg吐温20为表面活性剂,加入2 g 214型离子交换树脂,印迹时间为20 min。此工艺条件下制备得到的固定化印迹脂肪酶在反应中的酯化效果最高,固定化印迹酶的酯化效果是游离酶的4.35倍,固定化印迹酶催化酯化反应合成L-抗坏血酸棕榈酸酯后,最大产物浓度约为15.58 g?L^(-1),最佳转化率可达63.3%。This study combined bio-imprinting and enzyme immobilization methods to simplify thepreparation process of immobilized bio-imprinted enzyme.The production conditions such as enzyme solutionpH,imprinting molecule(olive oil)percentage,reaction time,supporter type of ion-exchange resin wasoptimized.The results show that the best conditions are pH=8.0,100mg olive oil,1mL ethanol as cosolvent,100mg Tween-20as surfactant,20min reaction time and2g type214ion-exchange resin.Immobilizedbio-imprinted enzymes with the highest esterification reaction efficiency can be obtained under such conditions,which is4.35times higher than free enzymes.The conversion of ascorbic acid is63.3%and the concentrationof L-ascorbyl palmitate is15.58g.L-1after esterification.

关 键 词：脂肪酶 生物印迹 固定化 酯化反应 L-抗坏血酸棕榈酸酯 

分 类 号：Q814.2[生物学—生物工程]