研究热敏载体的靶间距对固定化青霉素 G 酰化酶活性恢复的影响。

Study of target spacing of thermo-sensitive carrier on the activity recovery of immobilized penicillin G acylase.

机构信息

State Key Laboratory of Gansu Advanced Non-ferrous Metal Materials, Lanzhou University of Technology, Lanzhou, Gansu, 730050, China; School of Material Science and Engineering, Lanzhou University of Technology, Lanzhou, Gansu, 730050, China.

出版信息

Colloids Surf B Biointerfaces. 2019 Jul 1;179:153-160. doi: 10.1016/j.colsurfb.2019.03.064. Epub 2019 Apr 1.

DOI:10.1016/j.colsurfb.2019.03.064

PMID:30954878

Abstract

The immobilized penicillin G acylase (PGA) is an important industrial catalyst, the activity recovery rate of it directly affects enterprise efficiency. How to improve the enzyme activity recovery rate has been a research focus in this field. Based on the above problems, this work further improved the activity recovery rate by adjusting the target spacing for the first time. Glycidyl methacrylate (GMA) was used as the immobilized target and methyl methacrylate (MMA) as the copolymer monomer. According to the copolymer composition equation of P(MMA-co-GMA), the thermo-sensitive copolymers, PDEA-b-PHEMA-b-P(MMA-co-GMA) with different target spacings, were synthesized rapidly and efficiently via reversible addition-fragmentation chain transfer (RAFT) polymerization method. The error range between the theoretical and actual values of MMA and GMA in the copolymers carrier was (0-4)%, which demonstrated that the reliability of using composition equation to accurately and quickly synthesize copolymers with specific spacing. Studies on the thermo-sensitive showed that the low critical solution temperature (LCST) of the copolymer carrier decreased with the increase of hydrophobic monomer. Most importantly, the activity recovery rate increased with the increase of target spacing, and when the molar ratio of MMA to GMA in the copolymer was 8.75:1, the recovery of activity of immobilized PGA could be up to 63.50%, which was 21.70% higher than that of pure GMA. This work provided an important idea for improving the activity of immobilized PGA.

摘要

固定化青霉素 G 酰化酶（PGA）是一种重要的工业催化剂，其活性回收率直接影响企业效率。如何提高酶的活性回收率一直是该领域的研究重点。基于上述问题，本工作首次通过调整固定化靶间距进一步提高了酶的活性回收率。以甲基丙烯酸缩水甘油酯（GMA）为固定化靶，甲基丙烯酸甲酯（MMA）为共聚单体。根据共聚物组成方程 P（MMA-co-GMA），通过可逆加成-断裂链转移（RAFT）聚合方法快速高效地合成了不同靶间距的温敏共聚物 PDEA-b-PHEMA-b-P（MMA-co-GMA）。共聚物载体中 MMA 和 GMA 的理论值与实际值之间的误差范围在（0-4）%之间，表明使用组成方程准确快速合成具有特定间距的共聚物的可靠性。对温敏性的研究表明，共聚物载体的低临界溶液温度（LCST）随着疏水性单体的增加而降低。最重要的是，活性回收率随着靶间距的增加而增加，当共聚物中 MMA 与 GMA 的摩尔比为 8.75:1 时，固定化 PGA 的活性回收率可达到 63.50%，比纯 GMA 提高了 21.70%。这项工作为提高固定化 PGA 的活性提供了一个重要思路。