L-Alanine promotes anti-infectious properties of Bacillus subtilis S-2 spores via the germination receptor gerAA.

Publisher: Springer Country of Publication: United States NLM ID: 101484100 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1867-1314 (Electronic) Linking ISSN: 18671306 NLM ISO Abbreviation: Probiotics Antimicrob Proteins Subsets: MEDLINE

Original Publication: New York, NY. : Springer

Bacillus subtilis*/physiology ; Spores, Bacterial* ; Alanine*/pharmacology ; Bacterial Proteins*/genetics ; Bacterial Proteins*/metabolism ; Probiotics*/pharmacology; Humans ; Animals ; Caco-2 Cells ; Swine ; Epithelial Cells/microbiology ; Epithelial Cells/drug effects ; Cell Line

Bacillus species, which have two cell-type forms (vegetative cells and spores), demonstrate a variety of probiotic functions in animal feed additives and human nutrition. We previously found that the probiotic effect of Bacillus subtilis S-2 spores with high germination response to L-alanine was specifically enhanced by the L-alanine pretreatment. The germination response of Bacillus is highly associated with the germination receptors of spores. However, how L-alanine-induced germination of spores exerts anti-infectious effect in epithelial cells remains unclear. In this study, we constructed the mutant strain of B. subtilis S-2 with germination receptor gerAA knockout to further explore the role of spore germination in resisting pathogen infection to cells. The differential probiotic effects of B. subtilis S-2 and S-2 ^ΔgerAA spores pretreated with L-alanine were evaluated in intestinal porcine epithelial cells (IPEC-J2) or Caco2 cells infected with enterotoxigenic Escherichia coli (ETEC) or following IL-1β stimulation. The results showed that the germination response of the S-2 ^ΔgerAA spores to L-alanine was significantly reduced. Compared with the S-2 ^ΔgerAA spores, the L-alanine-induced germination of B. subtilis S-2 spores significantly increased the activity of anti-adhesion of ETEC to IPEC-J2 cells and reduced the expression of inflammatory factors and cell receptors. L-alanine induction also significantly promoted the expression of autophagy-related proteins in the B. subtilis S-2 spores. These findings demonstrate that the gerAA germination receptor is essential for the probiotic function of Bacillus spores and that L-alanine treatment promotes the anti-infectious properties of the germinated spores in porcine intestinal epithelial IPEC-J2 cells. The result suggests the importance of germination receptor gerAA in helping spore germination and enhancing anti-infectious activity. The findings in the study benefit to screening of potential Bacillus probiotics and increasing probiotic efficacy induced by L-alanine as an adjuvant.
(© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

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HZY23010 University-Enterprise R&D Cooperation Projects; HZY21089 University-Enterprise R&D Cooperation Projects; 32072767 and 31672455 National Natural Science Foundation of China; CZY21001, CZP22003 Fundamental Research Funds for the Central Universities South-Central MinZu University

Keywords: Bacillus; Germination receptor gerAA; IPEC-J2; L-Alanine; Spores

OF5P57N2ZX (Alanine)
0 (Bacterial Proteins)

Date Created: 20230713 Date Completed: 20240813 Latest Revision: 20240924

20240924

10.1007/s12602-023-10121-2

37439954