title: Site-Specific and High-Loading Immobilization of Proteins by Using Cohesin-Dockerin and CBM-Cellulose Interactions.
abstract: Immobilization of enzymes enhances their properties for application in industrial processes as reusable and robust biocatalysts. Here, we developed a new immobilization method by mimicking the natural cellulosome system. A group of cohesin and carbohydrate-binding module (CBM)-containing scaffoldins were genetically engineered, and their length was controlled by cohesin number. To use green fluorescent protein (GFP) as an immobilization model, its C-terminus was fused with a dockerin domain. GFP was able to specifically bind to scaffoldin via cohesin-dockerin interaction, while the scaffoldin could attach to cellulose by CBM-cellulose interaction. Our results showed that this mild and convenient approach was able to achieve site-specific immobilization, and the maximum GFP loading capacity reached ∼0.508 μmol/g cellulose.
authors.affiliation: Beijing Key Laboratory of Bioprocess. College of Life Science and Technology, Beijing University of Chemical Technology , Beisanhuan East Road #15, Beijing, China 100029.
authors.forename: Mei
authors.initials: M
authors.lastname: Li
authors.affiliation: Beijing Key Laboratory of Bioprocess. College of Life Science and Technology, Beijing University of Chemical Technology , Beisanhuan East Road #15, Beijing, China 100029.
authors.forename: Yi
authors.initials: Y
authors.lastname: Yue
authors.affiliation: Beijing Key Laboratory of Bioprocess. College of Life Science and Technology, Beijing University of Chemical Technology , Beisanhuan East Road #15, Beijing, China 100029.
authors.forename: Zi-Jian
authors.initials: ZJ
authors.lastname: Zhang
authors.affiliation: Beijing Key Laboratory of Bioprocess. College of Life Science and Technology, Beijing University of Chemical Technology , Beisanhuan East Road #15, Beijing, China 100029.
authors.forename: Zai-Yu
authors.initials: ZY
authors.lastname: Wang
authors.affiliation: Beijing Key Laboratory of Bioprocess. College of Life Science and Technology, Beijing University of Chemical Technology , Beisanhuan East Road #15, Beijing, China 100029.
authors.forename: Tian-Wei
authors.initials: TW
authors.lastname: Tan
authors.affiliation: Beijing Key Laboratory of Bioprocess. College of Life Science and Technology, Beijing University of Chemical Technology , Beisanhuan East Road #15, Beijing, China 100029.
authors.forename: Li-Hai
authors.initials: LH
authors.lastname: Fan
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D018797:Cell Cycle Proteins / Q000737:chemistry*; D002482:Cellulose / Q000737:chemistry*; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000737:chemistry; D055767:Immobilized Proteins / Q000737:chemistry*; D008958:Models, Molecular; D011487:Protein Conformation; D000072417:Protein Domains; D000097722:Cohesins
chemical_list: D001426:Bacterial Proteins; D018797:Cell Cycle Proteins; D002482:Cellulose; D002868:Chromosomal Proteins, Non-Histone; D049452:Green Fluorescent Proteins; D055767:Immobilized Proteins; D000097722:Cohesins