|
A. S. Dotsenko, A. M. Rozhkova, A. V. Gusakov
Properties
and N-glycosylation of recombinant endoglucanase II from Penicillium
verruculosum
Abstract
Cellulose is the major component of plant cell walls and the main
substrate in processes of biotransformation of lignocellulosic materials into
sugars and then into commercially valid organic compounds. Thus cellulases are
the major components of enzyme complexes used to hydrolyze cellulose into
sugars. Endoglucanase II (EGII) from Penicillium verruculosum was cloned
into Penicillium canescens. The recombinant EGII was isolated in
homogeneous form, and its properties were studied in comparison with a native
enzyme. Mass-spectrometry analysis was used to identify the N-glycosylation
sites and the structures of N-linked glycans. Both the native and recombinant
forms of EGII demonstrated similar biochemical and catalytic properties as well
as N-glycosylation patterns. N-linked high-mannose glycans and the products of
their enzymatic trimming, according to the formula (Man)1-9(GlcNAc)2,
were found at two N-glycosylation sites (N42 and N194) of both EGII forms.
Glycosylation at the third potential site (N19) was not detected.
Key words: cellulase,
endoglucanase, Penicillium verruculosum, cloning, N-glycosylation.
Copyright (C) Chemistry Dept., Moscow State University, 2002
|
|
