Abstract: Mold causes significant economic losses and threatens human and animal health. Iturin A is a natural compound with potent antifungal activity, and iturin A-producing bacteria can provide potential biocontrol solution for mold contamination. In this study, the biosynthesis of iturin A was significantly enhanced by deleting nonessential protease genes, enhancing ATP supply, and modifying the itu operon in Bacillus amyloliquefaciens. Accordingly, the efficient iturin A-producing strain B. amyloliquefaciens BAX-bac3 was constructed, and the iturin A yield reached 1.20 g/L, increased by 2.99-fold compared to the wild-type strain HZ-12. In vitro agar plate assays showed that the increased iturin A synthesis significantly enhanced the antifungal activity of BAX-bac3 against Penicillium chrysogenum and Penicillium expansum, two common pathogenic fungi. Additionally, biocontrol experiments conducted on tobacco leaves and apples further confirmed that BAX-bac3 treatment effectively prevented fungal infection. In conclusion, this study presents a novel and effective strategy to enhance iturin A synthesis for development of biocontrol agents.
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