LAUSR

Bacillus subtilis, a spore-forming industrial bacteria, is widely used for enzymes and valuable chemicals production. The spore-formation, however, always results in remarkably reduced cell-density, thereby reduced product-yield. Here, we construct different patterns of non-spore-forming B. subtilis via just single-gene regulation. During the three spore-forming stages: signal sensing, transduction and sporulation, we find that only deleting the single gene in the sporulation, i.e. spo0A, spoIIIE, and spoIVB, can completely block-up the spore-generation. Interestingly, the constructed non-sporulating mutants exhibited physiological heterogeneity and distinct synthetic capabilities. The spo0A-null spore-free mutant has great advantage on enzyme production, e.g. 194% enhancement for amylase production; whereas the spoIVB-null non-spore-forming mutant does very well in producing secondary metabolites, such as surfactin, its titer in flask highly increases to 16.7 g/L together with the overexpression and Leu addition strategy. Our results offer a new strategy for re-molding B. subtilis to further improve fermentation and application efficiency.