title: Immobilized trypsin on hydrophobic cellulose decorated nanoparticles shows good stability and reusability for protein digestion.
abstract: The preparation of biocatalysts based on immobilized trypsin is of great importance for both proteomic research and industrial applications. Here, we have developed a facile method to immobilize trypsin on hydrophobic cellulose-coated silica nanoparticles by surface adsorption. The immobilization conditions for the trypsin enzyme were optimized. The as-prepared biocatalyst was characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, and elemental analysis. In comparison with free enzyme, the immobilized trypsin exhibited greater resistances against thermal inactivation and denaturants. In addition, the immobilized trypsin showed good durability for multiple recycling. The general applicability of the immobilized trypsin for proteomic studies was confirmed by enzymatic digestion of two widely used protein substrates: bovine serum albumin (BSA) and cytochrome c. The surface adsorption protocols for trypsin immobilization may provide a promising strategy for enzyme immobilization in general, with great potential for a range of applications in proteomic studies.
authors.affiliation: College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China.
authors.forename: Xiuxia
authors.initials: X
authors.lastname: Sun
authors.affiliation: College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China.
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Cai
authors.affiliation: College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China; Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China. Electronic address: wangrq@lzu.edu.cn.
authors.forename: Ren-Qi
authors.initials: RQ
authors.lastname: Wang
authors.affiliation: College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China. Electronic address: xiaojx@lzu.edu.cn.
authors.forename: Jianxi
authors.initials: J
authors.lastname: Xiao
mesh_terms: D000818:Animals; D002417:Cattle; D002482:Cellulose / Q000737:chemistry*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions; D053758:Nanoparticles / Q000737:chemistry*; D059748:Proteolysis; D040901:Proteomics; D012822:Silicon Dioxide / Q000737:chemistry; D013696:Temperature; D014357:Trypsin / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D012822:Silicon Dioxide; D002482:Cellulose; D014357:Trypsin
keywords: Biocatalyst; Hydrophobic cellulose; Nanoparticle; Trypsin immobilization