title: Surface immobilization of protein via biosilification catalyzed by silicatein fused to glutathione S-transferase (GST).
abstract: 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.
authors.affiliation: Department of Biotechnology and Bioinformatics, Korea University, Jochiwon, Sejong 339-700, Korea.
authors.forename: Mi-Ran
authors.initials: MR
authors.lastname: Ki
authors.forename: Ki Baek
authors.initials: KB
authors.lastname: Yeo
authors.forename: Seung Pil
authors.initials: SP
authors.lastname: Pack
mesh_terms: D000818:Animals; D055162:Biocatalysis; D002403:Cathepsins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D004800:Enzymes, Immobilized / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012822:Silicon Dioxide / Q000378:metabolism; D047233:Suberites / Q000201:enzymology / Q000235:genetics
chemical_list: D004800:Enzymes, Immobilized; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D012822:Silicon Dioxide; D006735:Horseradish Peroxidase; D005982:Glutathione Transferase; D002403:Cathepsins