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AbrB knockout not positively affecting bacillomycin D production in Bacillus amyloliquefaciens fmbJ analyzed by
Jing Sun1, Shanshan Jin1, Yujuan Liu1
1College of Food Science and Engineering/Collaborative, Innovation Center for Modern Grain Circulation and Safety/Key Laboratory of Grains and Oils Quality Control and Processing, Nanjing University of Finance and Economics, Nanjing 210023, China.
Abstract:
Bacillomycin D (BD) is a potent lipopeptide with broad application potential encoded by bmyDABC with amino acids as substrates. To clarify the influence of transcriptional repressor AbrB regulating the synthesis of BD, abrB gene in Bacillus amyloliquefaciens fmbJ was successfully deleted by marker free method. Compared with the wild-type strain, BD production initially increased and then decreased. Expression of BD synthesis genes (bmyA-D) and signaling genes (spo0A, spo0B, and spo0E) all showed a brief rise in the early stage and a collective decline in the later stage, with sustained depletion of precursor amino acids (Tyr, Pro, Glu, Thr) after 36 h. EMSA and DNase I footprinting demonstrated that AbrB does not directly bind BD regulatory regions, while metabolomics revealed the exhaustion of fatty acid and amino acid precursors. These results showed that abrB knockout does not positively affecting BD production, providing guidance for more effective engineering of BD overproducing strains.
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