| Summary: | Derivatives of CenA and of CBD.PT from C. fimi with tandemly repeated binding
domains were constructed. A sensitive assay using 14C-labeled proteins was developed to
measure adsorption affinities for microcrystalline cellulose at low protein concentrations. It
was found that the CBDs in the double binding domain derivatives could function in tandem
to increase the overall adsorption affinity of CenA, provided that the catalytic domain was
present and that the CBDs were separated by a full Pro-Thr linker. The catalytic domain was
found to contribute to the overall adsorption affinity of the enzyme, as the affinity of the
isolated binding domain was reduced compared to that of CenA. Furthermore, the nature of
the linker separating domains in wild-type CenA was important for adsorption affinity of the
enzyme.
The activities of the double binding domain derivatives of CenA on microcrystalline
and soluble substrates were unchanged compared to the activity of the wild type enzyme.
Furthermore, double binding domain derivatives of CBD.PT released more small particles
from intact cotton fibres than did CBD.PT, and this effect was independent of the adsorption
affinity of the proteins. === Science, Faculty of === Microbiology and Immunology, Department of === Graduate
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