CIESC Journal ›› 2020, Vol. 71 ›› Issue (9): 4246-4254.DOI: 10.11949/0438-1157.20200509

• Biochemical engineering and technology • Previous Articles     Next Articles

Immobilization of alcohol dehydrogenase on the surface of polyethylene membrane

Mengya GUO1(),Shuxin JI1,Fengjuan GU1,Zihui MENG1,Wenfang LIU1(),Yanzi WANG2   

  1. 1.School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing 102488, China
    2.College of Chemistry and Environmental Engineering, Shenzhen University, Shenzhen 518060, Guangdong, China
  • Received:2020-05-08 Revised:2020-07-03 Online:2020-09-05 Published:2020-09-05
  • Contact: Wenfang LIU

醇脱氢酶在聚乙烯膜表面的固定化研究

郭梦雅1(),季书馨1,谷凤娟1,孟子晖1,刘文芳1(),王燕子2   

  1. 1.北京理工大学化学与化工学院,北京 102488
    2.深圳大学化学与环境工程学院,广东 深圳 518060
  • 通讯作者: 刘文芳
  • 作者简介:郭梦雅(1997—),女,硕士研究生,3293113722@qq.com
  • 基金资助:
    北京市自然科学基金项目(2172050);中国博士后基金项目(2019M650212)

Abstract:

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/PAA-PE. 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 9.6 μmol/(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 15.58 μmol/(L·min), 53.8% activity was remained after 10 cycles.

Key words: alcohol dehydrogenase, polyethylene film, polyethylenimine, enzyme immobilization, covalently bonding

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9.6 μmol/(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15.58 μmol/(L·min);重复利用10次后可保持53.8%的活性。

关键词: 醇脱氢酶, 聚乙烯膜, 聚乙烯亚胺, 酶固定化, 共价结合

CLC Number: