is a prevalent foodborne pathogen that induces food poisoning symptoms such as vomiting and diarrhea. Its capacity to form spores and biofilm enables it to withstand disinfectants and antimicrobials, leading to persistent contamination during food processing. Consequently, it is necessary to develop novel and efficient antimicrobial agents to control , its spores, and biofilms. Peptidoglycan hydrolases have emerged as a promising and eco-friendly alternative owing to their specific lytic activity against pathogenic bacteria. Here, we identified and characterized a Lysozyme-like cell wall hydrolase Lys14579, from the genome of ATCC 14579. Recombinant Lys14579 specifically lysed without affecting other bacteria. Lys14579 exhibited strong lytic activity against , effectively lysing cell within 20 min at low concentration (10 μg/mL). It also inhibited the germination of spores and prevented biofilm formation at 12.5 μg/mL. Moreover, Lys14579 displayed good antimicrobial stability with negligible hemolysis in mouse red blood cells and no cytotoxicity against RAW264.7 cells. Notably, Lys14579 effectively inhibited in boiled rice and minced meat in a dose-dependent manner. Furthermore, bioinformatics analysis and point mutagenesis experiments revealed that Glu-47 was the catalytic site, and Asp-57, Gln-60, Ser-61 and Glu-63 were active-site residues related with the cell wall lytic activity. Taken together, Lys14579 could be a promising biocontrol agent against vegetative cells, spores, and biofilm of in food industry.

中文翻译：

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对蜡状芽孢杆菌营养细胞、孢子和生物膜具有高活性的细胞壁水解酶的表征

是一种常见的食源性病原体，可引起呕吐和腹泻等食物中毒症状。它形成孢子和生物膜的能力使其能够耐受消毒剂和抗菌剂，从而导致食品加工过程中的持续污染。因此，有必要开发新型有效的抗菌剂来控制其孢子和生物膜。肽聚糖水解酶因其对病原菌的特异性裂解活性而成为一种有前景且环保的替代品。在这里，我们从 ATCC 14579 的基因组中鉴定并鉴定了一种溶菌酶样细胞壁水解酶 Lys14579。重组 Lys14579 可以特异性裂解而不影响其他细菌。Lys14579 对 表现出强烈的裂解活性，在低浓度 (10 μg/mL) 下可在 20 分钟内有效裂解细胞。12.5 μg/mL 时，它还能抑制孢子萌发并防止生物膜形成。此外，Lys14579 显示出良好的抗菌稳定性，在小鼠红细胞中溶血可忽略不计，并且对 RAW264.7 细胞没有细胞毒性。值得注意的是，Lys14579 以剂量依赖性方式有效抑制煮米和肉末。此外，生物信息学分析和点突变实验表明，Glu-47是催化位点，Asp-57、Gln-60、Ser-61和Glu-63是与细胞壁裂解活性相关的活性位点残基。综上所述，Lys14579 可能成为食品工业中针对营养细胞、孢子和生物膜的一种有前途的生物防治剂。