A streptavidin-cellulose-binding domain fusion protein that binds biotinylated proteins to cellulose.
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.
Le
K D
KD
Department of Microbiology, University of British Columbia, Vancouver, Canada.
Gilkes
N R
NR
Kilburn
D G
DG
Miller
R C
RC
Saddler
J N
JN
Warren
R A
RA
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*
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