畜産物利用2

[V-16-26] Functional enhancement and evaluation of ribosome-engineered Limosilactobacillus ingluviei C37 from broiler chicken

*ジャミヤンプレブ ソヨルマー¹、生井 楓¹、重盛 駿¹、下里 剛士¹ (1. 信州大バイオメディカル研)

Ribosome engineering (RE) is an approach that enhances microbial potential using antibiotics to induce spontaneous mutations in the ribosome. In this study, we applied RE in Limosilactobacillus ingluviei C37 (LIC37) from broiler chicken and evaluated the characteristics of the resultant mutants (MT) along with the impact of RE. The LIC37 wild-type parent (WT) was cultured on MRS agar plates containing streptomycin to obtain streptomycin-resistant MT. SDS-PAGE analysis and CBB staining were applied to the supernatant of the MT. We found that a band at approximately 200 kDa, which was later detected as cell wall anchor LPXTG-motif-containing-proteins (LPXTG), accumulated to significantly higher levels in MT_K57N compared to WT. RNA sequencing result showed increased accumulation of the transcripts encoding the LPXTG in MT_K57N too. However, the expression of sortase, an enzyme that anchors surface proteins to the cell wall, was decreased in the MT. We suggest that, in MT_K57N, the decreased expression of sortase leads to the release of LPXTG from the cell wall, resulting in the increased secretion and accumulation of these proteins in the culture supernatant.