Description of a cellulose-binding domain and a linker sequence from Aspergillus fungi.
A family I cellulose-binding domain (CBD) and a serine- and threonine-rich linker peptide were cloned from the fungi Aspergillus japonicus and Aspergillus aculeatus. A glutathione S-transferase (GST) fusion protein comprising GST and a peptide linker with the CBD fused to its C-terminus, was expressed in Escherichia coli. The renatured GST-CBD recovered from inclusion bodies had a molecular mass of 36.5 kDa which agrees with the 29 kDa of the GST plus the calculated 7.5 kDa of the linker with the CBD. The isolated GST-CBD protein adsorbed to both bacterial microcrystalline cellulose and carboxymethyl cellulose. Deletion of the linker peptide caused a decrease in cellulose adsorbance and a higher sensitivity to protease digestion.
Quentin
M
M
Department of Fibre and Paper Technology, ATO BV, PO Box 17, 6700 AA Wageningen, The Netherlands. m.g.e.quentin@ato.wag-ur.nl
Ebbelaar
M
M
Derksen
J
J
Mariani
C
C
van Der Valk
H
H
D000595:Amino Acid Sequence; D001230:Aspergillus / Q000737:chemistry* / Q000235:genetics; D002482:Cellulose / Q000378:metabolism*; D003001:Cloning, Molecular; D003460:Crystallization; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012694:Serine / Q000737:chemistry; D013912:Threonine / Q000737:chemistry
D011993:Recombinant Fusion Proteins; D013912:Threonine; D012694:Serine; D002482:Cellulose; C109691:microcrystalline cellulose