食窦魏斯氏菌可促进肠道健康,增强宿主免疫应答,为选育出耐酸性强的食窦魏斯氏菌,该研究以从泡菜中筛选出的食窦魏斯氏菌W25为出发菌株,通过紫外诱变(ultraviolet,UV,60 s)、甲基磺酸乙酯(ethyl methylsulfone,EMS)诱变(0.5%,45 min)及复合诱变UV-EMS和EMS-UV筛选获得U20、E7、U-E18和E-U28四株突变株。U-E18表现最优,产酸活力较W25显著提升;模拟胃肠环境耐受性,U-E18在pH 1.5胃液中存活率为20.92%,肠液环境中8 h存活率为13.78%,W25均完全失活;U-E18在8 h胆盐中生长率达32.48%,远高于W25的18.36%。功能基因组学分析揭示了U-E18的耐酸分子机制,蛋白质直系同源簇(cluster of orthologous groups of protein,COG)功能注释分析显示碳水化合物转运代谢、翻译及核糖体生物合成、氨基酸转运代谢等通路显著富集;KEGG通路分析鉴定出ABC转运蛋白系统具有关键作用。综上所述,U-E18为耐酸性强的优良菌株资源,为食品发酵的研究与应用提供了重要的候选菌株,具有广阔的开发潜力和应用前景。
Weissella cibaria demonstrates beneficial effects on gut health and the host immune response.To select acid-tolerant strains of W.cibaria, in this study, W.cibaria W25 screened from kimchi products was used as the starting strain.Through ultraviolet mutagenesis (UV, 60 s), ethyl methanesulfonate mutagenesis (EMS, 0.5%, 45 min), as well as composite mutagenesis, including sequential treatments:UV followed by EMS (UV-EMS) and EMS followed by UV (EMS-UV), four mutants, namely U20, E7, U-E18 and E-U28, were obtained.U-E18 showed the best performance, and its acid production activity was significantly higher than that of W25.In the simulation of the tolerance to the gastrointestinal environment, the survival rate of U-E18 in gastric fluid with a pH of 1.5 was 20.92%, and the survival rate in the intestinal fluid environment after 8 hours was 13.78%, W25 was completely inactivated.The growth rate of U-E18 in the presence of bile salts after 8 hours reached 32.48%, which was much higher than the 18.36% of W25.Functional genomics analysis revealed the molecular mechanism of acid tolerance of U-E18.Functional annotation analysis based on the cluster of orthologous groups of proteins (COG) revealed significant enrichment in pathways related to carbohydrate transport and metabolism, translation and ribosome biogenesis, as well as amino acid transport and metabolism.KEGG pathway analysis identified the ABC transporter system as playing a pivotal role.These findings establish U-E18 as an exceptional acid-tolerant microbial resource, offering substantial potential for food fermentation research and industrial applications.
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