Preparation of biodegradable and thermoresponsive enzyme-polymer conjugates with controllable bioactivity via RAFT polymerization

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• 作者：Huihui Liu ; Jizhen Zhang ; Xiong Luo ; Na Kong ; Liang Cui ; Jingquan Liu
• 关键词：Glucose oxidase ; Enzymatic activity ; RAFT polymerization ; Biodegradable ; Thermoresponsive
• 刊名：European Polymer Journal
• 出版年：2013
• 出版时间：October, 2013
• 年：2013
• 卷：49
• 期：10
• 页码：2949-2960
• 全文大小：1409 K

文摘

The preparation of biodegradable and thermoresponsive enzyme-polymer bioconjugates with controllable enzymatic activity via reversible addition?fragmentation chain transfer (RAFT) polymerization and amidation conjugation reaction is presented. A new 2-mercaptothiazoline ester functionalized RAFT agent with intra-disulfide linkage was synthesized and used as chain transfer agent (CTA) to generate a biocompatible homopolymer, poly(ethyleneglycol) acrylate (polyPEG-A) and a thermoresponsive copolymer of poly(ethyleneglycol) acrylate with di(ethyleneglycol)ethyl ether acrylate [poly(PEG-A-co-DEG-A)]. These biodegradable and thermoresponsive polymers were then conjugated to the surface of glucose oxidase (GOx) under mild condition to afford the biodegradable and thermoresponsive enzyme-polymer conjugates. Cleavage of the polymer chains from the GOx surface obviously recovered the enzymatic activity. The thermoresponsive test of GOx-poly(PEG-A-co-DEG-A) revealed that the bioconjugate exhibited regular enzymatic activity fluctuation upon the temperature change below or above the lower critical solution temperature (LCST). The as-prepared enzyme-polymer conjugates were also characterized using ¹H NMR, UV-vis spectroscopy, polyacrylamide gel electrophoresis (PAGE) and biocatalytic activity tests. These smart enzyme-polymer conjugates would envision promising applications in biotechnology and biomedicine.