Liu Yaping, Zhao Xiaoxiao, Yang Miao, Song Xin, Wang Guangqiang, Xia Yongjun, Zhao Liang, Xiong Zhiqiang, Ai Lianzhong
Shanghai Engineering Research Center of Food Microbiology, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China.
College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
Foods. 2025 May 16;14(10):1766. doi: 10.3390/foods14101766.
is a widely used probiotic with significant health benefits, but its application is limited by oxygen sensitivity. Our laboratory previously developed an oxygen-tolerant AR668-R1 using adaptive laboratory evolution under aerobic culture, but the molecular mechanism remains unclear. In this work, compared to the wild-type parental strain AR668, 212 upregulated and 390 downregulated proteins were identified in AR668-R1 under aerobic conditions through comparative proteomic analysis. Enrichment analysis of the differentially expressed proteins between AR668 and AR668-R1 identified the potential oxygen-tolerant related pathways, including the translation process, transmembrane transport system, and carbohydrate metabolism. Furthermore, five potential oxygen-tolerance proteins (DapE, Mth2, MutT, Eno, and MsrAB) were validated by RT-qPCR that may contribute to the aerobic growth of AR668-R1. Through gene overexpression validation, Mth2 (7,8-dihydro-8-oxoguanine triphosphatase) was found to enhance the growth of AR668-R1 by 19.8% compared to the empty plasmid control under aerobic conditions. Our finding provides valuable insights into the oxygen-tolerant mechanisms of at the protein level.
是一种广泛使用且具有显著健康益处的益生菌，但其应用受到氧敏感性的限制。我们实验室之前在需氧培养条件下通过适应性实验室进化开发出了耐氧的AR668-R1，但分子机制仍不清楚。在这项工作中，通过比较蛋白质组学分析，与野生型亲本菌株AR668相比，在需氧条件下AR668-R1中鉴定出212种上调蛋白和390种下调蛋白。对AR668和AR668-R1之间差异表达蛋白的富集分析确定了潜在的耐氧相关途径，包括翻译过程、跨膜运输系统和碳水化合物代谢。此外，通过RT-qPCR验证了五种潜在的耐氧蛋白（DapE、Mth2、MutT、Eno和MsrAB），它们可能有助于AR668-R1的需氧生长。通过基因过表达验证，发现在需氧条件下，与空质粒对照相比，Mth2（7,8-二氢-8-氧代鸟嘌呤三磷酸酶）使AR668-R1的生长提高了19.8%。我们的发现为 在蛋白质水平上的耐氧机制提供了有价值的见解。 （注：原文中“at the protein level”前似乎少了个具体的益生菌名称）
