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Appl Environ Microbiol. 1989 November; 55(11): 2739-2744
Copyright © 1989, American Society for Microbiology. All Rights Reserved.

Inducible Secretion of a Cellulase from Clostridium thermocellum in Bacillus subtilis

Gwennael Joliff, Alex Edelman, Andre Klier and Georges Rapoport*

Unité de Biochimie Microbienne, Département des Biotechnologies, URA 1300 CNRS, Institut Pasteur, 75724 Paris Cedex 15, France

ABSTRACT

A host-vector system for inducible secretion during the logarithmic growth phase in Bacillus subtilis has been developed. The B. subtilis levansucrase gene promoter and the region encoding its signal sequence have been used. The endoglucanase A of Clostridium thermocellum was used as a model protein to test the efficiency of the system. Effective inducible secretion of the endoglucanase A was observed when either the levansucrase signal sequence or its own signal sequence was used. Expression of the endoglucanase A in different genetic backgrounds of B. subtilis showed that its regulation was similar to that of levansucrase, and high enzyme activity was recovered from the culture supernatant of a hyperproducing B. subtilis sacU(Hy) strain. The molecular weight of 46,000 estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the secreted endoglucanase A is compatible with the calculated molecular weight of the mature polypeptide.

FOOTNOTES

* Corresponding author.

Appl Environ Microbiol. 1989 November; 55(11): 2739-2744
Copyright © 1989, American Society for Microbiology. All Rights Reserved.



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Copyright © 1989 by the American Society for Microbiology. All rights reserved.