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辣根过氧化物酶固定在β-环糊精包覆的银纳米颗粒上:其在生物传感器应用中的未来前景。

Immobilization of horseradish peroxidase on β-cyclodextrin-capped silver nanoparticles: Its future aspects in biosensor application.

作者信息

Karim Zoheb, Khan Mohd Jahir, Maskat Mohamad Yusof, Adnan Rohana

机构信息

a Division of Wood Science and Nanocomposite, Department of Material Science , Lulea University of Technology , Lulea , Sweden.

b School of Chemical Sciences and Food Technology, Faculty of Science and Technology , Universiti Kebangsaan Malaysia , Bangi , Selangor , Malaysia.

出版信息

Prep Biochem Biotechnol. 2016 May 18;46(4):321-7. doi: 10.1080/10826068.2015.1031389.

Abstract

This study aimed to work out a simple and high-yield procedure for the immobilization of horseradish peroxidase on silver nanoparticle. Ultraviolet-visible (UV-vis) and Fourier-transform infrared spectroscopy and transmission electron microscopy were used to characterize silver nanoparticles. Horseradish peroxidase was immobilized on β-cyclodextrin-capped silver nanoparticles via glutaraldehyde cross-linking. Single-cell gel electrophoresis (Comet assay) was also performed to confirm the genotoxicity of silver nanoparticles. To decrease toxicity, silver nanoparticles were capped with β-cyclodextrin. A comparative stability study of soluble and immobilized enzyme preparations was investigated against pH, temperature, and chaotropic agent, urea. The results showed that the cross-linked peroxidase was significantly more stable as compared to the soluble counterpart. The immobilized enzyme exhibited stable enzyme activities after repeated uses.

摘要

本研究旨在制定一种简单且高产的方法,用于将辣根过氧化物酶固定在银纳米颗粒上。利用紫外可见(UV-vis)光谱、傅里叶变换红外光谱和透射电子显微镜对银纳米颗粒进行表征。通过戊二醛交联将辣根过氧化物酶固定在β-环糊精包覆的银纳米颗粒上。还进行了单细胞凝胶电泳(彗星试验)以确认银纳米颗粒的遗传毒性。为降低毒性,用β-环糊精包覆银纳米颗粒。针对pH、温度和离液剂尿素,对可溶性和固定化酶制剂进行了稳定性比较研究。结果表明,与可溶性对应物相比,交联过氧化物酶的稳定性显著更高。固定化酶在重复使用后仍表现出稳定的酶活性。

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