Gastrointestinal tolerance enhancement of the LPxTG‐motif surface protein overexpressed Lactobacillus reuteri SH23in vivo

BACKGROUND Lactobacillus reuteri is a well‐recognized probiotic with numerous health benefits, including modulation of gut microbiota, anti‐inflammatory effects, and immune system enhancement. However, its probiotic efficacy depends largely on survival through the gastrointestinal tract and effective colonization of the gut mucosa. Surface‐anchored LPxTG‐motif proteins play crucial roles in bacterial adhesion and enhancing stress resistance. This study aimed to investigate the effects of LPxTG overexpression in L. reuteri SH23 on its gastrointestinal tolerance, adhesion capacity, and ability to regulate host intestinal flora. RESULTS A genetically modified strain, LPxTG‐pMG36e‐ L .SH23, exhibited a 2.42‐fold increase in LPxTG gene expression compared to wild‐type and control strains. While growth and acid production remained unchanged, the modified strain showed significantly enhanced cell surface hydrophobicity and self‐aggregation. Importantly, it demonstrated increased survival in simulated gastric (6.90 log CFU mL −1 ) and intestinal (5.78 log CFU mL −1 ) fluids. Adhesion to Caco‐2 cells improved markedly, reaching 19%. In a murine model, LPxTG‐pMG36e‐ L .SH23 reduced intestinal inflammation and significantly improved gut microbial diversity and abundance, suggesting enhanced colonization and probiotic function. CONCLUSION Overexpression of LPxTG‐motif proteins in L . reuteri SH23 strengthens its probiotic potential by improving gastrointestinal tolerance, promoting mucosal adhesion, and modulating intestinal microbiota. These findings support the functional importance of LPxTG surface proteins in probiotic–host interactions and suggest their value in the development of next‐generation probiotics with enhanced health benefits and gut persistence.