Affiliation:
1. Department of Microbiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada
Abstract
Both the periplasmic and the extracellular cellodextrinases from
Bacteroides succinogenes
S85 grown on Avicel microcrystalline cellulose were purified to homogeneity by column chromatography and characterized. Over 70% of the total cellobiosidase activity displayed by cells was accounted for by these enzymes. The periplasmic and extracellular cellodextrinases had identical molecular weights (50,000), as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and identical isoelectric points (4.9). In addition, the two enzymes were similar in catalytic properties, with
K
m
and
V
max
values of approximately 0.24 mM and 21 μmol/min per mg of protein, respectively. Examination of the two enzymes by using peptide mapping and immunoblotting techniques provided additional evidence indicating their identical nature. Immunoblotting of the extracellular culture fluid with affinity-purified antibody to the periplasmic cellodextrinase revealed one band with a molecular weight corresponding to that of the periplasmic cellodextrinase. The stability of the purified periplasmic cellodextrinase in aqueous solution was markedly enhanced by increased protein content. This enzyme showed a low affinity for crystalline cellulose.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
50 articles.
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