Ribosome-Engineered Lacticaseibacillus rhamnosus Strain GG Exhibits Cell Surface Glyceraldehyde-3-Phosphate Dehydrogenase Accumulation and Enhanced Adhesion to Human Colonic Mucin

Abstract

Differences in individual host responses have emerged as an issue in the health benefits of probiotics. Here, we applied ribosome engineering (RE) technology, developed in an actinomycetes study, to Lacticaseibacillus rhamnosus GG (LGG). RE can effectively enhance microbial potential by using antibiotics to induce spontaneous mutations in the ribosome and/or RNA polymerase. In this study, we identified eight types of streptomycin-resistant mutations in the LGG rpsL gene, which encodes ribosomal protein S12. Notably, LGG harboring the K56N mutant (LGG-MTK56N) expressed high levels of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) on the cell surface compared with the LGG wild-type (LGG-WT). GAPDH plays a key role in colon mucin adhesion. Indeed, LGG-MTK56N significantly increased type-A human colonic mucin adhesion compared to LGG-WT in experiments using the Biacore system. The ability to adhere to the colon is an important property of probiotics; thus, these results suggest that RE is an effective breeding strategy for probiotic lactic acid bacteria. IMPORTANCE We sought to apply ribosome engineering (RE) to probiotic lactic acid bacteria and to verify RE's impact. Here, we showed that one mutant of RE-Lacticaseibacillus rhamnosus GG (LGG-MTK56N) wore a GAPDH on the cell surface; the GAPDH was exported via an ABC transporter. Compared to the wild-type parent, LGG-MTK56N adhered more strongly to human colonic mucin and exhibited a distinct cell size and shape. These findings demonstrate that RE in LGG-MTK56N yielded dramatic changes in protein synthesis, protein transport, and cell morphology and affected adherence to human colonic mucin.

Development of an intestinal epithelial cell line and organoids derived from the same swine and characterization of their antiviral responses

“…The mucosal layer serves as the primary interface between the intestinal tract and the members of the microbiota or probiotics before direct contact with the intestinal epithelium. For example, Lacticaseibacillus rhamnosus GG, the most studied probiotic lactic acid bacterium, is known to bind to mucin via glyceraldehyde-3-phosphate dehydrogenase (GAPDH) on its cell wall surface, and it has been reported that the binding strength varies depending on the amount of GAPDH [26]. Furthermore, the mucin within the intestinal mucosa not only functions as a scaffold for microbial adhesion but also serves as a nutrient source for bacterial proliferation [27,28].…”

Glyceraldehyde-3-Phosphate Dehydrogenase Increases the Adhesion of Lactobacillus reuteri to Host Mucin to Enhance Probiotic Effects

“…Mucin functions as an adhesion receptor for surface GAPDH in lactobacilli [ 38 , 40 ]. Adhesion of lactobacilli to the intestine is partly due to the binding of GAPDH to human ABO-type blood group antigens expressed on human colonic mucin [ 41 ].…”

Oral administration of Brevibacterium linens from washed cheese increases the proportions of short‐chain fatty acid‐producing bacteria and lactobacilli in the gut microbiota of mice

“…An overnight culture of B. linens was centrifuged at 8,000× g for 5 min at 4°C. The obtained bacterial pellets were observed under a JSM 7600‐F SEM (JEOL, Tokyo, Japan) according to previously reported methods (Ishida et al, 2020; Tsukagoshi et al, 2020).…”