Characterization of germinants and their receptors for spores of non-food-borne Clostridium perfringens strain F4969.
Clostridium perfringens type A can cause both food poisoning (FP) and non-food-borne (NFB) gastrointestinal diseases. Our previous study reported that a mixture of l-asparagine and KCl (AK)-germinated spores of FP and NFB isolates well, but KCl and, to a lesser extent, l-asparagine induced spore germination only in FP isolates. We now report that the germination response of FP and NFB spores differsignificantly in several defined germinants and rich media. Spores of NFB strain F4969 gerAA, gerKA-KC or gerKC mutants lacking specific germinant receptor proteins germinated more slowly than wild-type spores with rich media, did not germinate with AK and germinated poorly compared to wild-type spores with l-cysteine. The germination defects in the gerKA-KC spores were largely due to loss of GerKC as (i) gerKA spores germinated significantly with all tested germinants, while gerKC spores exhibited poor or no germination; and (ii) germination defects in gerKC spores were largely restored by expressing the wild-type gerKA-KC operon in trans. We also found that gerKA-KC, gerAA and gerKC spores, but not gerKA spores, released dipicolinic acid at a slower rate than wild-type spores with AK. The colony-forming efficiency of F4969 gerKC spores was also ~35-fold lower than that of wild-type spores, while gerAA and wild-type spores had similar viability. Collectively, these results suggest that the GerAA and GerKC proteins play roles in normal germination of C. perfringens NFB isolates and that GerKC, but not GerAA, is important in these spores' apparent viability.
Department of Biomedical Sciences, Oregon State University, Corvallis, OR 97331, USA.|Department of Microbiology, Oregon State University, Corvallis, OR 97331, USA.|Medical Laboratories Department, College of Science Al-Zulfi, Majmaah University, Al Majmaah, Saudi Arabia.
Saeed
S
Banawas
Department of Biomedical Sciences, Oregon State University, Corvallis, OR 97331, USA.|Gut Microbiota and Gastrointestinal Disease Research Group, Departamento de Ciencias Biológicas, Facultad de Ciencias Biológicas, Universidad Andres Bello, Santiago, Chile.
Daniel
D
Paredes-Sabja
Department of Molecular, Microbial and Structural Biology, UConn Health, Farmington, CT 06030, USA.
Peter
P
Setlow
Department of Microbiology, Oregon State University, Corvallis, OR 97331, USA.|Department of Biomedical Sciences, Oregon State University, Corvallis, OR 97331, USA.
Mahfuzur R
MR
Sarker
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D003016:Clostridium perfringens / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism; D050296:Microbial Viability; D013171:Spores, Bacterial / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism
D001426:Bacterial Proteins; D003545:Cysteine