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Applied and Environmental Microbiology, August 2005, p. 4577-4584, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4577-4584.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Mycosubtilin Overproduction by Bacillus subtilis BBG100 Enhances the Organism's Antagonistic and Biocontrol Activities

Valérie Leclère,1 Max Béchet,1 Akram Adam,2 Jean-Sébastien Guez,1 Bernard Wathelet,3 Marc Ongena,2 Philippe Thonart,2 Frédérique Gancel,1 Marlène Chollet-Imbert,1 and Philippe Jacques1*

Laboratory of Microbial Bioprocesses, Polytech'Lille, University of Science and Technology of Lille, F-59655 Villeneuve d'Ascq Cedex, France,1 Centre Wallon de Biologie Industrielle, University of Liege, B40, B-4000 Liège, Belgium,2 Unité de Chimie Biologique Industrielle, Agricultural University of Gembloux, B-5030 Gembloux, Belgium3

Received 27 October 2004/ Accepted 28 February 2005

A Bacillus subtilis derivative was obtained from strain ATCC 6633 by replacement of the native promoter of the mycosubtilin operon by a constitutive promoter originating from the replication gene repU of the Staphylococcus aureus plasmid pUB110. The recombinant strain, designated BBG100, produced up to 15-fold more mycosubtilin than the wild type produced. The overproducing phenotype was related to enhancement of the antagonistic activities against several yeasts and pathogenic fungi. Hemolytic activities were also clearly increased in the modified strain. Mass spectrometry analyses of enriched mycosubtilin extracts showed similar patterns of lipopeptides for BBG100 and the wild type. Interestingly, these analyses also revealed a new form of mycosubtilin which was more easily detected in the BBG100 sample. When tested for its biocontrol potential, wild-type strain ATCC 6633 was almost ineffective for reducing a Pythium infection of tomato seedlings. However, treatment of seeds with the BBG100 overproducing strain resulted in a marked increase in the germination rate of seeds. This protective effect afforded by mycosubtilin overproduction was also visualized by the significantly greater fresh weight of emerging seedlings treated with BBG100 compared to controls or seedlings inoculated with the wild-type strain.

* Corresponding author. Mailing address: Laboratory of Microbial Bioprocesses (LABEM), Polytech'Lille, University of Science and Technology of Lille, Avenue du Professeur Langevin, F-59655 Villeneuve d'Ascq Cedex, France. Phone: 33 3 28 76 74 40. Fax: 33 3 28 76 74 01. E-mail: philippe.jacques{at}polytech-lille.fr.

Applied and Environmental Microbiology, August 2005, p. 4577-4584, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4577-4584.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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