Functional starch-binding domain of Aspergillus glucoamylase I in Escherichia coli.
We have fused three starch-binding domains (SBD) encoding gene fragments (residues 511-616, 495-616 and 481-616) of glucoamylase I (GAI) to the 3' end of the Escherichia coli malE gene encoding maltose-binding protein (MBP). The fusion proteins were produced in E. coli and were purified by chromatography on cross-linked amylose. Factor Xa digestion of the fusion proteins resulted in the release of functional SBD fragments which were separated from MBP on the basis of their differential binding to cross-linked amylose. The amino acid (aa) composition of the purified SBD fragments agreed with the respective amino acid compositions of GAI. The sizes of the SBD fragments were 11.9, 13.8 and 15.6 kDa, respectively.
Department of Food Science and Human Nutrition, Iowa State University, Ames 50011.
A R
AR
Kusnadi
C
C
Ford
Z L
ZL
Nikolov
D018528:ATP-Binding Cassette Transporters; D000595:Amino Acid Sequence; D001230:Aspergillus / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008320:Maltose / Q000378:metabolism; D058619:Maltose-Binding Proteins; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins; D009838:Oligodeoxyribonucleotides; D033902:Periplasmic Binding Proteins; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping
D018528:ATP-Binding Cassette Transporters; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C461619:MalE protein, E coli; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D009838:Oligodeoxyribonucleotides; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins; C461599:maltose transport system, E coli; D008320:Maltose; D005087:Glucan 1,4-alpha-Glucosidase