title: A streptavidin-cellulose-binding domain fusion protein that binds biotinylated proteins to cellulose.
abstract: A fusion protein, Sta-CBDCex, which comprises streptavidin with a cellulose-binding domain (CBDCex) fused to its C terminus, was produced in the cytoplasm of Escherichia coli, where it formed inclusion bodies. Renatured Sta-CBDCex, recovered from the inclusion bodies, adsorbed to Avicel, a microcrystalline cellulose. The cellulose-bound Sta-CBDCex in turn bound biotinylated alkaline phosphatase or biotinylated beta-glucosidase. The immobilized beta-glucosidase remained fully active during 2 weeks of continuous column operation at 50 degrees C.
authors.affiliation: Department of Microbiology, University of British Columbia, Vancouver, Canada.
authors.forename: K D
authors.initials: KD
authors.lastname: Le
authors.forename: N R
authors.initials: NR
authors.lastname: Gilkes
authors.forename: D G
authors.initials: DG
authors.lastname: Kilburn
authors.forename: R C
authors.initials: RC
authors.lastname: Miller
authors.forename: J N
authors.initials: JN
authors.lastname: Saddler
authors.forename: R A
authors.initials: RA
authors.lastname: Warren
mesh_terms: D001710:Biotin; D015153:Blotting, Western; D002482:Cellulose; D003001:Cloning, Molecular / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004800:Enzymes, Immobilized; D004926:Escherichia coli; D044643:Glucan 1,4-beta-Glucosidase; D006026:Glycoside Hydrolases / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D043325:Xylan Endo-1,3-beta-Xylosidase; D001617:beta-Glucosidase / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D011993:Recombinant Fusion Proteins; D001710:Biotin; D002482:Cellulose; D006026:Glycoside Hydrolases; D001617:beta-Glucosidase; D043325:Xylan Endo-1,3-beta-Xylosidase; D044643:Glucan 1,4-beta-Glucosidase