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Functional Immobilization of a Biofilm-Releasing Glycoside Hydrolase Dispersin B on Magnetic Nanoparticles
Applied Biochemistry and Biotechnology ( IF 3.1 ) Pub Date : 2021-09-15 , DOI: 10.1007/s12010-021-03673-y
Zewen Liu 1 , Zisong Zhao 1 , Kai Zeng 1 , Yue Xia 1 , Weihua Xu 2 , Ruoyu Wang 2 , Junhui Guo 1 , Hao Xie 1
Affiliation  

Dispersin B (DspB) is a member of glycoside hydrolase family 20 (GH20) and catalyzes degradation of biofilms forming by pathogenic bacteria such as Staphylococcus aureus. Magnetoreceptor (MagR) is a magnetic protein that can be used as a fusion partner for functionally immobilizing proteins on magnetic surfaces. In the present study, a recombinant protein DspB-MagR was constructed by fusing MagR to the C-terminus of DspB and expressed in Escherichia coli. Magnetic immobilization of purified DspB-MagR on magnetic core–shell structured Fe3O4@SiO2 nanoparticles was achieved and characterized by means of various techniques including SDS-PAGE, Fourier transform infrared spectroscopy, thermogravimetric analysis, zeta potential measurement, and scanning electron microscopy. It was evaluated the influence of temperature, pH, and storage time on the performance of immobilized DspB-MagR on Fe3O4@SiO2 nanoparticles. Removal of biofilms forming by Staphylococcus aureus and other medical sourced bacterial species was achieved by using Fe3O4@SiO2 nanoparticles loading with DspB-MagR. This work promoted potential applications of DspB and similar enzymes for medical purposes.



中文翻译:


生物膜释放糖苷水解酶分散素 B 在磁性纳米颗粒上的功能固定化



分散素 B (DspB) 是糖苷水解酶家族 20 (GH20) 的成员,可催化金黄色葡萄球菌等病原菌形成的生物膜降解。磁感受器 (MagR) 是一种磁性蛋白,可用作将蛋白质功能性固定在磁性表面上的融合伴侣。本研究通过将MagR融合到DspB的C端构建了重组蛋白DspB-MagR,并在大肠杆菌中表达。通过SDS-PAGE、傅里叶变换红外光谱、热重分析、zeta电位测量和扫描电子等多种技术,实现了纯化的DspB-MagR在磁性核壳结构Fe 3 O 4 @SiO 2纳米颗粒上的磁固定化和表征显微镜。评估了温度、pH和储存时间对Fe 3 O 4 @SiO 2纳米颗粒上固定化DspB-MagR性能的影响。通过使用负载有 DspB-MagR 的 Fe 3 O 4 @SiO 2纳米粒子来去除由金黄色葡萄球菌和其他医用来源的细菌物种形成的生物膜。这项工作促进了 DspB 和类似酶在医学上的潜在应用。

更新日期:2021-09-15
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