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O. G. Korotkova, M. V. Semenova, E. A. Rubtsova, O. A. Sinitsyna, E. G. Kondrat`eva, N. M. Bibikov, A. M. Rozhkova, A. P. Sinitsyn
Comparative analysis of the properties of
recombinant endo-1,4-β-glucanase ii and its
chimeric form with a cellular binding module
Abstract
A chimeric form of Penicillium
verruculosum endo-1,4-β-glucanase II (EGII),
to the C-terminus of which contains a cellulose-binding module (CBM) of P.
verruculosum cellobiohydrolase I, has produced by genetic engineering
method. In native form the egII gene does not have a region encoding
CBM. The resulting chimeric enzyme was isolated in a homogeneous form and its
properties has been studied. The addition of CBM to EGII led to significant
increase in the activity of the chimeric enzyme to microcrystalline cellulose
(MCC), as well as its adsorption capacity for cellulose has been observed.
However, the addition of CBM to EGII led to a decrease in activity towards
soluble polysaccharide substrates (carboxymethylcellulose and β-glucan). The chimeric form of the enzyme in the composition of the
cellulase complex allowed to hydrolyze cellulose-containing substrates more
effectively. The increase in reducing sugars (RS) in the case of MCC hydrolysis
was 22% after 24 hours with a cellulose complex containing cellobiohydrolase I,
β-glucosidase and EGII-CBM in comparison to the
complex containing EGII without CBM.
Key words: endo-1,4-b-endoglucanase,
complex of cellulases, cellulose-binding module, hydrolysis, Penicillium
verruculosum.
Copyright (C) Chemistry Dept., Moscow State University, 2002
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