Abstract
Prodigiosin produced by marine bacterium Vibrio ruber DSM 14379 exhibits a potent antimicrobial activity against a broad range of Gram positive and Gram negative bacteria. The mechanism of prodigiosin antimicrobial action, however, is not known. In this work, the effect of prodigiosin on Bacillus subtilis growth, cell membrane leakage, and induction of autolysins was studied. Treating B. subtilis with prodigiosin resulted in rapid decline of optical density and increased cell membrane leakage measured by β-galactosidase activity. Cell lysis was initiated immediately after treatment with prodigiosin in the middle exponential phase and was completed within 2 h. Lytic activity of prodigiosin in mutant strains with impaired autolysin genes lytABCD decreased for 80% compared to the wild type strain, while in lytABCDEF mutant strain prodigiosin had no bacteriolytic but only bacteriostatic effect. Fast prodigiosin lytic activity on individual B. subtilis cells was confirmed by a modified comet assay. The results indicate that prodigiosin autolysin induction in B. subtilis is growth phase dependent.
Highlights
Prodigiosin, a secondary metabolite, is produced by several bacterial genera including Serratia, Streptomyces, Vibrio, Hahella, Zooshikella, and Pseudoalteromonas (Farmer et al, 1988; Sawabe et al, 1998; Shieh et al, 2003; Williamson et al, 2006; Kumar and Nair, 2007; Fehér et al, 2010; Staricet al., 2010; Lee et al, 2011; Stankovic et al, 2012)
Bacterial strains used in this study were B. amyloliquefaciens FZB42, B. licheniformis ATCC9445A, B. subtilis NCIB3610 all obtained from BGSC, B. subtilis ATCC6051, B. mycoides DSM2048 obtained from DSMZ, B. subtilis PS-216 wt (Stefanic and Mandic-Mulec, 2009), B. subtilis PS-216 amyE::PhyperspankmKate2 cat (Stefanic et al, 2015), Escherichia coli MG1655 and Vibrio ruber DSM14379 (Stopar et al, 2004; Boricet al., 2011)
The Minimal inhibitory concentration (MIC) values for different Bacillus sp. strains were in the range between 5 and 7 mg L−1 (MIC values for B. amyloliquefaciens FZB42 was cells treated in the late exponential phase; (C) cells treated in the stationary phase
Summary
Prodigiosin, a secondary metabolite, is produced by several bacterial genera including Serratia, Streptomyces, Vibrio, Hahella, Zooshikella, and Pseudoalteromonas (Farmer et al, 1988; Sawabe et al, 1998; Shieh et al, 2003; Williamson et al, 2006; Kumar and Nair, 2007; Fehér et al, 2010; Staricet al., 2010; Lee et al, 2011; Stankovic et al, 2012). It has affinity to DNA (Melvin et al, 2000), but shows no in vitro or in vivo genotoxic effect on Salmonella typhimurium cells (Guryanov et al, 2013). It modulates H+/Cl− symport activity (Konno et al, 1998). Prodigiosin has anticancer, antimalarial and immunosuppressant properties (Pérez-Tomás et al, 2003; Williamson et al, 2007) It inhibits growth of a wide range of Gram positive bacteria including Bacillus subtilis and Staphylococcus aureus, as well as Gram negative Escherichia coli, Salmonella enterica, and Erwinia carotovora (reviewed in Stankovic et al, 2014). We study the molecular targets of prodigiosin in B. subtilis
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