Cloning of Clostridium cellulovorans endo-1,4-beta-glucanase genes

Biochemical and Biophysical Research Communications
O ShoseyovR H Doi

Abstract

A Clostridium cellulovorans lambda gt11 gene bank was screened for endo-1,4-beta-glucanase [EC 3.2.1.4, EGase, Carboxy Methyl Cellulase (CMCase)] genes using a chromogenic substrate. Three genes (engA, engB, and engC) were isolated. The engB expressed the most active CMCase. The engA encoded a bifunctional enzyme that displayed endo-1,4-beta-glucanase and beta-glucosidase activities. The three recombinant glucanases, when expressed in Escherichia coli, were partially degraded into multiform active enzymes as evidenced by their SDS-PAGE-CMC zymograms. None of the clones could degrade crystalline cellulose, thus supporting the hypothesis that the integrity of the C. cellulovorans cellulase complex was essential for its 'true cellulase' activity.

Citations

Oct 23, 1997·Nature Structural Biology·J SakonP A Karplus
Mar 15, 2002·Chemical Record : an Official Publication of the Chemical Society of Japan ... [et Al.]·R H Doi, Y Tamaru
Nov 13, 2004·Applied Microbiology and Biotechnology·André O S LimaJoão L Azevedo
Jun 9, 2006·Microbiology and Molecular Biology Reviews : MMBR·Oded ShoseyovIlan Levy
Dec 15, 1994·FEMS Microbiology Letters·G T AttwoodB A White
Sep 27, 2000·Journal of Bacteriology·Y TamaruR H Doi
Dec 25, 2008·Chemical Record : an Official Publication of the Chemical Society of Japan ... [et Al.]·Edward A BayerHarry J Flint

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