A Small-Molecular-Weight Bacteriocin-like Inhibitory Substance (BLIS) UI-11 Produced by Lactobacillus plantarum HYH-11 as an Antimicrobial Agent for Aeromonas hydrophila

由植物乳杆菌HYH-11产生的小分子量细菌素样抑制物质(BLIS)UI-11可作为嗜水气单胞菌的抗菌剂

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Abstract

Aeromonas hydrophila is a typical pathogen that causes fish diseases and can easily infect different fish species. This study investigated the antibacterial activity, physicochemical properties and antibacterial mechanism of the BLIS UI-11 produced by Lactobacillus plantarum HYH-11, isolated from traditional kimchi in Hebei, China. It was found that BLIS UI-11 showed excellent inhibitory effect on the growth of A. hydrophila, and it also had a good antibacterial effect on various pathogens such as Vagococcus fluvialis, Listeria monocytogenes, Aeromonas dhakensis, Aeromonas salmonicida, Salmonella Typhimurium, Escherichia coli and Staphylococcus aureus. By measuring growth kinetics, it was found that the maximum antibacterial activity was reached after 30 h of culture, and both the optical density value at 600 nm (OD(600)) and pH basically entered the stable phase after 20 h. Whole-genome analysis and gene cluster prediction identified a RiPP-like biosynthetic gene cluster, which comprises genes encoding precursor peptides, modification enzymes, and transport/immunity components. The molecular weight of the antimicrobial active substance was detected by dialysis and Tricine-SDS-PAGE, and it was shown to be an ultra-small molecular substance (<1 kDa). BLIS UI-11 was sensitive to protease K, but its antibacterial activity remained stable after treatment with acidic environment (pH 3.0-6.0), high-temperature treatment (121 °C for 30 min), and ultraviolet irradiation (4 h). After the sub-live cell assay (PI/SYTO9) and scanning electron microscopy (SEM), BLIS UI-11 inhibited the growth of bacteria by destroying the cell membrane of A. hydrophila to deform, collapse, and form holes that lead to accounting leakage. The hemolysis assay indicated that BLIS UI-11 exhibited incomplete hemolysis, suggesting its safety for application. The results showed that BLIS UI-11 produced by strain HYH-11 has great potential as an antimicrobial agent against A. hydrophila infection.

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