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Title
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Overproduction, purification and characterization of the cellulose-binding domain of the Erwinia chrysanthemi secreted endoglucanase EGZ.
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Authors
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E.Brun,
P.Gans,
D.Marion,
F.Barras.
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Ref.
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Eur J Biochem, 1995,
231,
142-148.
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PubMed id
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Abstract
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EGZ is the major endoglucanase secreted by Erwinia chrysanthemi. Functional
characterization indicates that it is made of a catalytic N-terminal domain
linked to a C-terminal cellulose-binding domain (CBD) by a Ser/Thr-rich linker.
A chimeric plasmid, in which the CBD-encoding region was fused downstream of the
ompA signal sequence, was constructed and introduced into Escherichia coli. This
allowed for the production of processed and disulfide-bonded CBD, mostly
recovered from the culture supernatant of E. coli. One-dimensional NMR analysis
of the purified CBD reveals that it folds into a well-structured domain.
Moreover, comparison with the one-dimensional NMR analysis of full-length EGZ
strongly suggests that the CBD folds autonomously, providing experimental
support for the existence of domains of EGZ.
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