Purification and Comparison of the Periplasmic and Extracellular Forms of the Cellodextrinase from Bacteroides succinogenes.

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 mumol/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.

journal_name

Appl Environ Microbiol

authors

Huang L,Forsberg CW

doi

10.1128/AEM.54.6.1488-1493.1988

keywords:

subject

Has Abstract

pub_date

1988-06-01 00:00:00

pages

1488-93

issue

6

eissn

0099-2240

issn

1098-5336

journal_volume

54

pub_type

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