Surface immobilization of protein via biosilification catalyzed by silicatein fused to glutathione S-transferase (GST)
Surface immobilization of protein via biosilification catalyzed by silicatein fused to glutathione S-transferase (GST)
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DOI：
10.1007/s00449-012-0818-x
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发表时间：
2013-05-01
影响因子：
3.8
通讯作者：
Pack, Seung Pil
中科院分区：
文献类型：
--
作者：
Ki, Mi-Ran;Yeo, Ki Baek;Pack, Seung Pil
在近中性pH和环境温度条件下，已知来自土状钻的硅酸盐蛋白在体外催化二氧化硅沉积。本研究利用谷胱甘肽（GST）-谷胱甘肽（GSH）相互作用系统提高硅酸盐蛋白的产量，并发展了一种高效的蛋白固定化方法。在E.大肠杆菌中表达，GST-SIL蛋白在GSH包被的玻璃板上表达。GST-SIL结合的表面或基质可以在环境温度和中性pH下在正硅酸乙酯作为底物的存在下催化二氧化硅层的形成。在硅酸盐蛋白介导的硅化过程中，绿色荧光蛋白（GFP）或辣根过氧化物酶（HRP）可以有效地固定在二氧化硅表面上。固定化的GFP或HRP保持其活性并逐渐释放。这种生物相容性的二氧化硅涂层技术可用于制备生物分子固定化表面或基质，其可用于开发生物催化、诊断和生物传感系统或组织培养支架。
Silicatein from Suberites domuncula was known to catalyze silica deposition in vitro under near neutral pH and ambient temperature conditions. In this study, we employed GST-glutathione (GSH) interaction system to increase the production of silicatein and develop an efficient protein immobilization method. Recombinant silicatein fused with GST (GST-SIL) was produced in E. coli and the GST-SIL protein was employed on GSH-coated glass plate. GST-SIL bound surface or matrix can catalyze the formation of silica layer in the presence of tetraethyl orthosilicate as a substrate at an ambient temperature and neutral pH. During silicatein-mediated silicification, green fluorescent protein (GFP) or horseradish peroxidase (HRP) can be efficiently immobilized on the silica surface. Immobilized GFP or HRP retained their activity and were released gradually. This biocompatible silica coating technique can be employed to prepare biomolecule-immobilized surfaces or matrixes, which are useful for the development of biocatalytic, diagnostic and biosensing system, or tissue culture scaffolds.