title: Effect of the cortex-lytic enzyme SleC from non-food-borne Clostridium perfringens on the germination properties of SleC-lacking spores of a food poisoning isolate.
abstract: The hallmark of bacterial spore germination is peptidoglycan cortex hydrolysis by cortex-lytic enzymes. In spores of Clostridium perfringens wild-type strain SM101, which causes food poisoning, the sole essential cortex-lytic enzyme SleC is activated by a unique serine protease CspB. Interestingly, the non-food-borne wild-type strain F4969 encodes a significantly divergent SleC variant (SleCF4969) and 3 serine proteases (CspA, CspB, and CspC). Consequently, in this study we evaluated the functional compatibility of SleCF4969 and SleCSM101 by complementing the germination phenotypes of SM101ΔsleC spores with sleCF4969. Our results show that although pro-SleCF4969 was processed into mature SleCF4969 in the SM101ΔsleC spores, it partially restored spore germination with nutrient medium, with a mixture of ʟ-asparagine and KCl, or with a 1:1 chelate of Ca2+ and dipicolinic acid. While the amount of dipicolinic acid released was lower, the amount of hexosamine-containing material released during germination of SM101ΔsleC(sleCF4969) spores was similar to the amount released during germination of SM101 wild-type spores. The viability of SM101ΔsleC(sleCF4969) spores was 8- and 3-fold lower than that of SM101 and F4969 spores, respectively. Together, these data indicate that the peptidoglycan cortex hydrolysis machinery in the food poisoning isolate SM101 is functionally divergent than that in the non-food-borne isolate F4969.
authors.affiliation: Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Paredes-Sabja
authors.forename: Mahfuzur R
authors.initials: MR
authors.lastname: Sarker
mesh_terms: D003016:Clostridium perfringens / Q000201:enzymology* / Q000302:isolation & purification / Q000502:physiology; D005517:Foodborne Diseases / Q000382:microbiology; D005816:Genetic Complementation Test; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D010457:Peptidoglycan / Q000494:pharmacology; D010848:Picolinic Acids / Q000378:metabolism; D013171:Spores, Bacterial / Q000201:enzymology / Q000235:genetics / Q000254:growth & development
chemical_list: D010457:Peptidoglycan; D010848:Picolinic Acids; D006867:Hydrolases; C004860:dipicolinic acid