Multiple response optimization of Bacillus subtilis EA-CB0015 culture and identification of antifungal metabolites
Fecha
2014
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Biocatalysis and Agricultural Biotechnology
Resumen
The low yields of biomass and antimicrobial metabolites obtained in fermentation processes are limiting factors for implementing biological control agents in the field -- In this context, optimization of the culture medium for the biological control agent Bacillus subtilis EA-CB0015 was conducted in submerged culture to maximize the biomass production and antifungal activity -- Additionally, the active metabolites against the phytopathogen Mycosphaerella fijiensis produced under optimized conditions were identified -- Carbon and nitrogen constituents of the culture medium were optimized using a 22 factorial design with central point followed by a multiple response optimization coupled to the desirability function approach -- The optimized medium (33.4 g/L of glucose and 32.5 g/L of yeast extract), showed significant increases in both, cell density by 3.6 folds (13.271.0 g/L) and antifungal activity by 1.2 folds (77.770.3% inhibition) -- To gain insight into the type of active compounds, they were purified and identified by mass spectrometry (MS) -- The MS analysis revealed the presence of three families of lipopeptides: surfactin, fengycin and iturin -- The high antifungal activity was associated with the novel fengycin C and with iturin -- A which were partially growth-associated with a maximum concentration of 781.4 and 355.4 mg/L at 36 h of growth, respectively -- These results indicate the potential of B. subtilis EA-CB0015 to produce high concentrations of biomass and lipopeptides that can be exploited for biotechnological application