Differentiated pellicle organization and lipopeptide production in standing culture of Bacillus subtilis strains.

Pellicle formation and lipopeptide production was analysed in standing cultures of different Bacillus subtilis strains producing two or three families of lipopeptides. Despite its ability to produce surfactin, B. Subtilis ATCC 6633 was unable to form stable pellicle at air-water interface. For the ATTC 21332 and ATCC 9943 strains, it was shown for the first time that the lipopeptides were also produced in standing cultures at productivities similar or lower than those obtained when the culture medium is agitated. A differentiated behaviour was observed between these strains in repetitive batch cultures. B. subtilis 9943 formed a wrinkled, thinner and more resistant pellicle than B. subtilis 21332. The structure of the pellicle determined by electron microscopy observations showed that cells of B. subtilis 9943 formed microcolonies whereas those of B. subtilis 21332 rapidly died. Under these conditions, surfactin production by strain 21332 decreased after 2 days whereas it remained stable for B. subtilis 9943 during the 6 days of the cultures. These data indicate that cells of B. subtilis strains growing in pellicle can produce lipopeptides differently depending on their cellular organisation

Impact of energy supply and oxygen transfer on selective lipopeptide production by Bacillus subtilis BBG21.

The influence of power dissipation and volumetric oxygen transfer coefficient k(L)a on Bacillus subtilis productivity of lipopeptides surfactin and fengycin was studied in shake flasks in view of scaling-up of this fermentation process. The experiments performed with different flask sizes, relative filling volumes, and shaking frequencies confirmed clearly that lipopeptide production changed in function of power dissipation, via interfacial gas-liquid contact surface and oxygen supply. It was demonstrated that k(L)a is the key parameter controlling the productivity and the selectivity of the bioreaction. Varying the oxygen transfer conditions, the synthesis could be oriented to mixed production or to surfactin mono-production. The fraction of surfactin towards total lipopeptides produced and the maximal surfactin production both increased with k(L)a increase (surfactin concentration about 2 g L(-1) at k(L)a=0.04-0.08 s(-1)), while the maximal fengycin production (fengycin concentration about 0.3 g L(-1)) was obtained at moderate oxygen supply (k(L)a=0.01 s(-1))