title: Sortase-deficient lactobacilli: effect on immunomodulation and gut retention.
abstract: Surface proteins of probiotic microbes, including Lactobacillus acidophilus and Lactobacillus gasseri, are believed to promote retention in the gut and mediate host-bacterial communications. Sortase, an enzyme that covalently couples a subset of extracellular proteins containing an LPXTG motif to the cell surface, is of particular interest in characterizing bacterial adherence and communication with the mucosal immune system. A sortase gene, srtA, was identified in L. acidophilus NCFM (LBA1244) and L. gasseri ATCC 33323 (LGAS_0825). Additionally, eight and six intact sortase-dependent proteins were predicted in L. acidophilus and L. gasseri, respectively. Due to the role of sortase in coupling these proteins to the cell wall, ΔsrtA deletion mutants of L. acidophilus and L. gasseri were created using the upp-based counterselective gene replacement system. Inactivation of sortase did not cause significant alteration in growth or survival in simulated gastrointestinal juices. Meanwhile, both ΔsrtA mutants showed decreased adhesion to porcine mucin in vitro. Murine dendritic cells exposed to the ΔsrtA mutant of L. acidophilus or L. gasseri induced lower levels of pro-inflammatory cytokines TNF-α and IL-12, respectively, compared with the parent strains. In vivo co-colonization of the L. acidophilus ΔsrtA mutant and its parent strain in germ-free 129S6/SvEv mice resulted in a significant one-log reduction of the ΔsrtA mutant population. Additionally, a similar reduction of the ΔsrtA mutant was observed in the caecum. This study shows for the first time that sortase-dependent proteins contribute to gut retention of probiotic microbes in the gastrointestinal tract.
authors.affiliation: Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: Emma K
authors.initials: EK
authors.lastname: Call
authors.affiliation: Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: Yong Jun
authors.initials: YJ
authors.lastname: Goh
authors.affiliation: Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: Kurt
authors.initials: K
authors.lastname: Selle
authors.affiliation: Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: Todd R
authors.initials: TR
authors.lastname: Klaenhammer
authors.affiliation: Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: O'Flaherty
mesh_terms: D019881:Aminoacyltransferases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D018938:Caco-2 Cells; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D003713:Dendritic Cells / Q000276:immunology / Q000382:microbiology; D041981:Gastrointestinal Tract / Q000276:immunology / Q000382:microbiology*; D006801:Humans; D056747:Immunomodulation; D007778:Lactobacillus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D007779:Lactobacillus acidophilus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D051379:Mice; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
chemical_list: D001426:Bacterial Proteins; D014409:Tumor Necrosis Factor-alpha; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases