High-throughput screening and inhibition mechanism analysis of Bacillus licheniformis BL-4 as an antagonistic strain for Lactobacillus panis

Abstract

In Maotai-flavor Baijiu production, reducing lactic acid (LA) can alleviate microbial imbalance and flavor disharmony caused by LA accumulation. Current methods for reducing LA mainly focus on physical removal and fermentation parameter control, but they address only the symptoms, not the underlying cause. This study selected Lactobacillus panis antagonistic bacteria to control LA production at its source by inhibiting L. panis growth and analyzed its antimicrobial substances and mechanisms. Firstly, a high-throughput screening method for L. panis antagonists was developed based on lactate dehydrogenase, which correlates LA concentration with reduced nicotinamide adenine dinucleotide (NADH). Subsequently, a total of 34 antagonists were screened, with Bacillus licheniformis BL-4 exhibiting the highest inhibition rate (62.25%) against L. panis. Moreover, the primary antimicrobial substance, antimicrobial peptide antiL24, was purified from B. licheniformis BL-4 and evaluated for its activity and sequence. Finally, the mechanism of antiL24 against L. panis was analyzed by using microstructural analyses and transcriptomic profiling, revealing that antiL24 disrupts the cell wall and membrane of L. panis and affects genes involved in energy metabolism and protein synthesis. This study proposes a novel strategy for regulating LA concentration in Maotai-flavor Baijiu production, with the potential to enhance its quality.

Keywords

High-throughput screening / Bacillus licheniformis / Antimicrobial peptide / Lactic acid / Maotai-flavor Baijiu / Transcriptomic analysis / Biological Sciences / Microbiology

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Funding

Foundation for Innovative Research Groups of the National Natural Science Foundation of China(32021005); Fundamental Research Funds for the Central Universities(JUSRP124034); Jiangsu Basic Research Center for Synthetic Biology(BK20233003); Natural Science Foundation of Jiangsu Province(BK20202002)