摘要
以聚丙烯酸(PAA)改性的聚乙烯(PE)膜为载体,研究了醇脱氢酶(ADH)的两种固定化路线,并以甲醛为底物考察了固定化酶的催化性能。路线1用聚乙烯亚胺(PEI)进一步改性,使用戊二醛(GA)固定化ADH。最优固定化pH为6.0,温度为5~15℃,酶浓度为1.0 mg/ml,GA浓度为0.01%(质量);固定化酶的最适反应pH为6.5,温度为15~30℃,反应速率最高为9.6μmol/(L·min);重复利用10次后可保持47.3%的活性。路线2以PAA-PE为载体,用1-(3-二甲氨基丙基)-2-乙基碳二亚胺盐酸盐(EDC)和N-羟基琥珀酰亚胺(NHS)为活化剂,固定化ADH。EDC和NHS最优摩尔比为1∶0.5,固定化时间为24 h;固定化酶的最适反应pH为6.5,温度为20~37℃,反应速率为15.58μmol/(L·min);重复利用10次后可保持53.8%的活性。
Using polyacrylic acid(PAA)modified polyethylene(PE)membrane as a carrier,two immobilization routes of alcohol dehydrogenase(ADH)were studied,and the catalytic performance of immobilized enzyme was investigated using formaldehyde as a substrate.In the first route,PAA-PE membrane was further modified by polyethyleneimine(PEI)and then ADH was covalently linked by glutaraldehyde(GA)to the surface of PEI/PAAPE.The results show that the optimal immobilization pH was 6.0,immobilization temperature was 5-15℃,ADH and GA concentrations were 1.0mg/ml and 0.01%(mass).For immobilized enzyme,the optimal reaction pH was 6.5,temperature was 15-30℃,and the highest reaction rate was 9.6μmol/(L·min),the remaining activity was 47.3%after 10 use cycles.In the second route,ADH was immobilized on PAA-PE membrane with 1-(3-dimethylaminopropyl)-2-ethylcarbodiimide hydrochloride(EDC)and N-hydroxysuccinimide(NHS)as activators.The results show that the optimal molar ratio of EDC and NHS was 1∶0.5,and the immobilization time was 24 h.For immobilized enzyme,the optimal reaction pH was 6.5,temperature was 20-37℃,and the highest reaction rate was 15.58μmol/(L·min),53.8%activity was remained after 10 cycles.
作者
郭梦雅
季书馨
谷凤娟
孟子晖
刘文芳
王燕子
GUO Mengya;JI Shuxin;GU Fengjuan;MENG Zihui;LIU Wenfang;WANG Yanzi(School of Chemistry and Chemical Engineering,Beijing Institute of Technology,Beijing 102488,China;College of Chemistry and Environmental Engineering,Shenzhen University,Shenzhen 518060,Guangdong,China)
出处
《化工学报》
EI
CAS
CSCD
北大核心
2020年第9期4246-4254,共9页
CIESC Journal
基金
北京市自然科学基金项目(2172050)
中国博士后基金项目(2019M650212)。
关键词
醇脱氢酶
聚乙烯膜
聚乙烯亚胺
酶固定化
共价结合
alcohol dehydrogenase
polyethylene film
polyethylenimine
enzyme immobilization
covalently bonding
作者简介
第一作者:郭梦雅(1997-),女,硕士研究生,3293113722@qq.com;通信作者:刘文芳(1977-),女,博士,副教授,liuwenfang@bit.edu.cn。
