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酶固定于甲基丙烯酸2-羟乙酯和甲基丙烯酸缩水甘油酯共聚物刷上。

Immobilization of enzymes on 2-hydroxyethyl methacrylate and glycidyl methacrylate copolymer brushes.

作者信息

Ren Tanchen, Mao Zhengwei, Moya Sergio Enrique, Gao Changyou

机构信息

MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027 (China), Fax: (+86) 571-87951108.

出版信息

Chem Asian J. 2014 Aug;9(8):2132-9. doi: 10.1002/asia.201402150. Epub 2014 Jun 24.

Abstract

The immobilization of enzymes is of paramount importance to maintain their activity and stability. In this study, surface-initiated atom-transfer radical polymerization was applied to prepare poly(2-hydroxyethyl methacrylate)-block-poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) brushes on glass slides. The polymerization kinetics was followed by using a quartz crystal microbalance with dissipation monitoring and ellipsometry in terms of mass and thickness growth, respectively. The surface chemical compositions of the obtained polymer brushes were characterized by X-ray photoelectron spectroscopy. Their mass, thickness, and enzyme-immobilization ability could be easily tuned by the initiator reaction time, monomer ratio, and polymerization time. The antibacterial activity and stability of the immobilized lysozymes were studied by fluorescent staining and bacteria lysis assay, which revealed that the lysozymes on the copolymer brushes had good stability during storage at 4 °C for up to 30 days.

摘要

酶的固定化对于维持其活性和稳定性至关重要。在本研究中,采用表面引发原子转移自由基聚合方法在载玻片上制备聚(甲基丙烯酸2-羟乙酯)-嵌段-聚(甲基丙烯酸2-羟乙酯-共-甲基丙烯酸缩水甘油酯)刷。分别通过使用具有耗散监测功能的石英晶体微天平以及椭偏仪跟踪聚合动力学,以监测质量和厚度的增长。通过X射线光电子能谱对所得聚合物刷的表面化学成分进行表征。其质量、厚度和酶固定化能力可通过引发剂反应时间、单体比例和聚合时间轻松调节。通过荧光染色和细菌裂解试验研究了固定化溶菌酶的抗菌活性和稳定性,结果表明共聚物刷上的溶菌酶在4 °C下储存长达30天期间具有良好的稳定性。

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