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Description of a cellulose-binding domain and a linker sequence from Aspergillus fungi
M Quentin1, M Ebbelaar, J Derksen
1Department of Fibre and Paper Technology, ATO BV, PO Box 17, 6700 AA Wageningen, The Netherlands. m.g.e.quentin@ato.wag-ur.nl
Abstract:
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.
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