MassIVE MSV000085346 - Induced homogenous sporulation by kinA-overexpression leads to altered Bacillus subtilis spore proteomes and spore properties

To facilitate more accurate spore proteomic analysis, the current study focuses on inducing homogeneous sporulation by overexpressing kinA and assesses the effect of sporulation synchronization on spore resistance, structures, the germination behavior at single-spore level and the proteome. The results indicate that, in our set up, the synchronized sporulation by overexpressing kinA can generate a spore yield of 70% within 8 hours. The procedure increases spore wet heat resistance and thickness of the spore coat and cortex layers, whilst delaying the time to spore phase-darkening and burst after addition of germinant. The proteome analysis reveals that the upregulated proteins in the kinA induced spores, compared to spores without kinA induction as well as the wildtype spores, are mostly involved in spore formation. The downregulated proteins mostly belong to the categories of coping with stress, carbon and nitrogen metabolism, as well as regulation of sporulation. Thus, while kinA overexpression enhances sporulation synchronicity it also has profound effects on the central equilibrium of spore formation and spore germination through modulation of the spore molecular composition and stress resistance physiology.