1. Description of a cellulose-binding domain and a linker sequence from Aspergillus fungi.
- Author
-
Quentin M, Ebbelaar M, Derksen J, Mariani C, and van Der Valk H
- Subjects
- Amino Acid Sequence, Aspergillus genetics, Cloning, Molecular, Crystallization, Escherichia coli genetics, Escherichia coli metabolism, Molecular Sequence Data, Protein Binding, Protein Structure, Tertiary, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins metabolism, Serine chemistry, Threonine chemistry, Aspergillus chemistry, Cellulose metabolism
- 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.
- Published
- 2002
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