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A Bacillus subtilis derivative was obtained from strain ATCC 6633 byreplacement of the native promoter of the mycosubtilin operon by a constitutivepromoter originating from the replication gene repU of the Staphylococcusaureus plasmid pUB110. The recombinant strain, designated BBG100, produced upto 15-fold more mycosubtilin than the wild type produced. The overproducingphenotype was related to enhancement of the antagonistic activities againstseveral yeasts and pathogenic fungi. Hemolytic activities were also clearlyincreased in the modified strain. Mass spectrometry analyses of enrichedmycosubtilin extracts showed similar patterns of lipopeptides for BBG100 andthe wild type. Interestingly, these analyses also revealed a new form ofmycosubtilin which was more easily detected in the BBG100 sample. When testedfor its biocontrol potential, wild-type strain ATCC 6633 was almost ineffectivefor reducing a Pythium infection of tomato seedlings. However, treatment ofseeds with the BBG100 overproducing strain resulted in a marked increase in thegermination rate of seeds. This protective effect afforded by mycosubtilinoverproduction was also visualized by the significantly greater fresh weight ofemerging seedlings treated with BBG100 compared to controls or seedlingsinoculated with the wild-type strain.

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