Fusion to an endoglucanase allows alkaline phosphatase to bind to cellulose.
Endoglucanase CenA of Cellulomonas fimi comprises an N-terminal cellulose-binding domain and a C-terminal catalytic domain joined together by a sequence of 23 proline and threonine residues (the Pro-Thr box). The domains function independently when separated by proteolysis. TnphoA has been used to generate cenA'-'phoA fusions. CenA'-'PhoA fusion polypeptides which contain the entire cellulose-binding domain of CenA bind to cellulose, allowing their purification from periplasmic extracts in a single, facile step. This result has implications for purification or immobilisation of chimeric proteins on a cheap cellulose matrix.
Greenwood
J M
JM
Department of Microbiology, University of British Columbia, Vancouver, Canada.
Gilkes
N R
NR
Kilburn
D G
DG
Miller
R C
RC
Warren
R A
RA
D000192:Actinomycetales / Q000201:enzymology; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002480:Cellulase / Q000235:genetics / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology; D005374:Filtration / Q000295:instrumentation; D010209:Paper; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*
D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D002482:Cellulose; D000469:Alkaline Phosphatase; D002480:Cellulase