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Magnetic thermosensitive polymer composite carrier with target spacing for enhancing immobilized enzyme performance
Enzyme and Microbial Technology ( IF 3.4 ) Pub Date : 2021-08-19 , DOI: 10.1016/j.enzmictec.2021.109896
Hongyi Tu 1 , Boyuan Zhang 1 , Xiayun Zhang 1 , Chunli Zhao 1 , Lin Li 1 , Jianbin Wang 1 , Zhenbin Chen 1 , Pingbo Wang 1 , Zhizhong Li 2
Affiliation  

A novel magnetic thermosensitive polymer composite carrier with target spacing was developed. In this strategy, thermosensitive polymer grafted on magnetic Fe3O4 for enhancing immobilized penicillin G acylase (PGA) performance and introduce immobilized target spacing into magnetic carriers for the first time. Fe3O4 nanoparticles were synthesized by a reverse microemulsion method. The modifier used was the silane coupling agent γ-methylacryloxypropyl trimethoxysilane (KH570) and then reacting with a reversible-adaptive fragmentation chain transfer (RAFT) reagent, 2-cyano-2-propyldodecyl trithiocarbonate (CPDTC). The thermo-sensitive nanoparticle-composite carrier of Fe3O4-grafted-poly N, N-diethyl acrylamide-block-poly β-Hydroxyethyl methacrylate-block-random copolymer of glycidyl methacrylate and methyl methacrylate (Fe3O4-g-PDEA-b-PHEMA-b-P(MMA-co-GMA)) were synthesized by RAFT polymerization technique that used N, N-diethyl acrylamide (DEA), β-Hydroxyethyl methacrylate (HEMA), Glycidyl methacrylate (GMA) and Methyl methacrylate (MMA) as monomer, then which were employed as functional carriers for the immobilization of PGA. Within the carrier, the epoxy group of GMA segment was a target immobilization site for PGA and the introduction of MMA reflected the target space of immobilized PGA to improve catalytic activity and catalytic activity recovery rate of the immobilized PGA. Characterizations demonstrated that the triblock copolymers grafted Fe3O4 nanoparticles were successfully fabricated by the structure design. Besides, under these circumstances the enzyme activity (EA), enzyme loading capacity (ELC) and catalytic activity recovery ration (CAR) reached 31235 U/g, 128.39 mg/g and 93.32 %, respectively. The catalytic activity of immobilized PGA maintained 87.4 % of initial value and the recovery ratio (R) of immobilized PGA reached 96.22 % after recycling 12 times. Furthermore, the immobilized PGA exhibited advantages of low temperature homogeneous catalysis and magnetic separation, which indicated broad application prospects in the biocatalysts’ field.



中文翻译:

具有靶间距的磁性热敏聚合物复合载体提高固定化酶的性能

研制了一种新型的具有靶间距的磁性热敏聚合物复合载体。在该策略中,热敏聚合物接枝在磁性 Fe 3 O 4 上以增强固定化青霉素 G 酰基转移酶 (PGA) 的性能,并首次将固定化目标间距引入磁性载体。Fe 3 O 4纳米颗粒是通过反相微乳液法合成的。使用的改性剂是硅烷偶联剂 γ-甲基丙烯酰氧基丙基三甲氧基硅烷 (KH570),然后与可逆-自适应断裂链转移 (RAFT) 试剂、2-氰基-2-丙基十二烷基三硫代碳酸酯 (CPDTC) 反应。Fe 3 O 4热敏纳米颗粒复合载体-接枝聚 N, N-二乙基丙烯酰胺-嵌段聚甲基丙烯酸 β-羟乙酯-甲基丙烯酸缩水甘油酯和甲基丙烯酸甲酯的嵌段-无规共聚物 (Fe 3 O 4 - g -PDEA- b -PHEMA- b -P(MMA- co-GMA)) 以 N, N-二乙基丙烯酰胺 (DEA)、甲基丙烯酸 β-羟乙酯 (HEMA)、甲基丙烯酸缩水甘油酯 (GMA) 和甲基丙烯酸甲酯 (MMA) 为单体,通过 RAFT 聚合技术合成,然后用作官能团。用于固定 PGA 的载体。在载体内部,GMA链段的环氧基是PGA的目标固定位点,MMA的引入反映了固定化PGA的目标空间,提高了固定化PGA的催化活性和催化活性回收率。表征表明,通过结构设计成功地制备了接枝Fe 3 O 4纳米颗粒的三嵌段共聚物。此外,在这些情况下,酶活性 ( EA )、酶载量 (ELC )和催化活性回收率( CAR )分别达到31235 U/g、128.39 mg/g和93.32 %。固定化PGA的催化活性保持初始值的87.4%,循环12次后固定化PGA的回收率( R )达到96.22%。此外,固定化PGA表现出低温均相催化和磁分离的优点,在生物催化剂领域具有广阔的应用前景。

更新日期:2021-08-21
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