celB, a gene coding for a bifunctional cellulase from the extreme thermophile "Caldocellum saccharolyticum".

Abstract:

:"Caldocellum saccharolyticum" is an obligatory anaerobic thermophilic bacterium. A gene from this organism, designated celB, has been cloned in Escherichia coli as part of a bacteriophage lambda gene library. This gene produces a thermostable cellulase that shows both endoglucanase and exoglucanase activities on test substrates and is able to degrade crystalline cellulose to glucose. The sequence of celB has homology with both exo- and endoglucanases described by others. It appears to have a central domain without enzymatic activity which is joined to the enzymatic domains by runs of amino acids rich in proline and threonine (PT boxes). Deletion analysis shows that the exoglucanase activity is located in the amino-terminal domain of the enzyme and that endoglucanase activity is located in the carboxy-terminal domain. There are internal transcriptional and translational start sites within the gene. The intact gene has been cloned into a temperature-inducible expression vector, pJLA602, and overexpressed in E. coli. Polyacrylamide gel electrophoresis showed that celB produced a protein with a molecular weight of 118,000 to 120,000. A number of smaller proteins with activity against carboxymethyl cellulose and 4-methyl umbelliferyl-beta-D-cellobioside were also produced. These are believed to be the result of alternative translational start sites and/or proteolytic degradation products of the translated gene product.

journal_name

Appl Environ Microbiol

authors

Saul DJ,Williams LC,Grayling RA,Chamley LW,Love DR,Bergquist PL

doi

10.1128/AEM.56.10.3117-3124.1990

subject

Has Abstract

pub_date

1990-10-01 00:00:00

pages

3117-24

issue

10

eissn

0099-2240

issn

1098-5336

journal_volume

56

pub_type

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