This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Mingardon, F.
Right arrow Articles by Fierobe, H.-P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mingardon, F.
Right arrow Articles by Fierobe, H.-P.
Agricola
Right arrow Articles by Mingardon, F.
Right arrow Articles by Fierobe, H.-P.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, March 2005, p. 1215-1222, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1215-1222.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Heterologous Production, Assembly, and Secretion of a Minicellulosome by Clostridium acetobutylicum ATCC 824

Florence Mingardon,1 Stéphanie Perret,1 Anne Bélaïch,1 Chantal Tardif,1,2 Jean-Pierre Bélaïch,1,2 and Henri-Pierre Fierobe1*

BIP-CNRS,1 Université de Provence, Marseille, France2

Received 29 July 2004/ Accepted 19 October 2004

The gene man5K encoding the mannanase Man5K from Clostridium cellulolyticum was cloned alone or as an operon with the gene cipC1 encoding a truncated scaffoldin (miniCipC1) of the same origin in the solventogenic Clostridium acetobutylicum. The expression of the heterologous gene(s) was under the control of a weakened thiolase promoter Pthl. The recombinant strains of the solventogenic bacterium were both found to secrete active Man5K in the range of milligrams per liter. In the case of the strain expressing only man5K, a large fraction of the recombinant enzyme was truncated and lost the N-terminal dockerin domain, but it remained active towards galactomannan. When man5K was coexpressed with cipC1 in C. acetobutylicum, the recombinant strain secreted almost exclusively full-length mannanase, which bound to the scaffoldin miniCipC1, thus showing that complexation to the scaffoldin stabilized the enzyme. The secreted heterologous complex was found to be functional: it binds to crystalline cellulose via the carbohydrate binding module of the miniscaffoldin, and the complexed mannanase is active towards galactomannan. Taken together, these data show that C. acetobutylicum is a suitable host for the production, assembly, and secretion of heterologous minicellulosomes.

* Corresponding author. Mailing address: BIP-CNRS, 31, Chemin Joseph-Aiguier, F-13402 Marseille Cedex 20, France. Phone: 33 (0) 4 91 16 42 99. Fax: 33 (0) 4 91 71 33 21. E-mail: hpfierob{at}ibsm.cnrs-mrs.fr.

Applied and Environmental Microbiology, March 2005, p. 1215-1222, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1215-1222.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Shoseyov, O., Shani, Z., Levy, I. (2006). Carbohydrate Binding Modules: Biochemical Properties and Novel Applications. Microbiol. Mol. Biol. Rev. 70: 283-295 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»