Effect of pps disruption and constitutive expression of srfA on surfactin productivity, spreading and antagonistic properties of Bacillus subtilis 168 derivatives

• Published in: Journal of applied microbiology Vol. 109; no. 2; pp. 480 - 491
• Main Authors: Coutte, F, Leclère, V, Béchet, M, Guez, J.-S, Lecouturier, D, Chollet-Imbert, M, Dhulster, P, Jacques, P
• Format: Journal Article Web Resource
• Language: English
• Published: Oxford, UK Oxford, UK : Blackwell Publishing Ltd 01-08-2010; Blackwell Publishing Ltd; Blackwell; Oxford University Press; Wiley; Wiley-Blackwell
• Subjects: Antifungal activity; Bacillus subtilis; Bacillus subtilis - genetics; Bacillus subtilis - metabolism; Bacteriology; Biological and medical sciences; Biological control; Bioremediation; Biosynthesis; Biotechnology; Constitutive expression; Culture media; Disruption; Fatty Acids - biosynthesis; Fundamental and applied biological sciences. Psychology; Fungi - growth & development; Fungicides; Life Sciences; Lipopeptides - biosynthesis; Microbial Interactions; Microbiologie; Microbiology; Microbiology and Parasitology; Mutants; Oligopeptides - biosynthesis; Operon; Peptides, Cyclic - biosynthesis; Plant diseases; plipastatin; Productivity; Sciences du vivant; Sfp gene; spreading; srfA operon; Surfactin; Productivity; surfactin; Bacillus subtilis; Applied microbiology; Operon; spreading; Bacillaceae; Bacillales; constitutive expression; Bacteria; Antagonist; plipastatin; srfA operon
• ISSN: 1364-5072; 1365-2672; 1365-2672
• DOI: 10.1111/j.1365-2672.2010.04683.x

To analyse the effects of plipastatin operon disruption and constitutive expression of surfactin operon in Bacillus subtilis 168 on surfactin productivity, in vitro invasive growth and antagonism against fungi. The srfA native promoter was replaced by the constitutive promoter PrepU in B. subtilis 168 after integration of a functional sfp gene. Moreover, the plipastatin synthesis was further disrupted in the B. subtilis 168 derivatives. In liquid media, an earlier and higher expression of PrepU, than that found with PsrfA, led to a specific surfactin production fivefold higher after 6 h of culture. On solid media, not only the invasive growth and the haemolytic activity but also the antifungal activity of the constitutive strains were improved when compared to the parental strain BBG111. As expected, the disruption of the plipastatin operon strongly reduced in vitro antifungal properties but, interestingly, enhanced specific surfactin production (1·47 g g⁻¹ of biomass), spreading behaviour and haemolytic activity of the strains. This work demonstrates for the first time the interdependency of surfactin and plipastatin regarding their biosynthesis as well as their influence on the biological activities of the producing strain. The constitutive overproduction of surfactin enhances the invasive growth and the in vitro antagonistic activity of the mutant strain. Both properties are known to play an important role in the biocontrol of plant diseases. Plipastatin operon disruption increases the surfactin productivity of mutant strains. These mutants are interesting for use in continuous bioprocesses for surfactin production or in bioremediation.