title: Purification and Characterization of a Rabbit Serum Factor That Kills Listeria Species and Other Foodborne Bacterial Pathogens.
abstract: In an in-vitro assay, rabbit serum, but not human serum, killed Listeria monocytogenes, a foodborne pathogen. The aim of our study was to purify and partially characterize this killing factor. Listericidin was purified from rabbit serum by a single-step ion-exchange chromatography with DEAE-Sephadex A-50 and its antimicrobial activity was assessed by a microdilution method. Listericidin is a protein with a molecular weight of 9 kDa and an isoelectric point of 8.1. It kills L. monocytogenes at 4°C, 25°C, and 37°C, and its activity is resistant to heat (boiling) and acidic conditions (pH <2). Listericidin's activity is inhibited by sodium chloride and various growth media, is sensitive to proteolytic enzymes and is enhanced by calcium chloride, and is neutralized by monoclonal antibodies to human complement C3a. However, the listericidin reacts weakly with these antibodies in an ELISA. The first 33 N-terminal residues of listericidin (SVQLTEKRMDKVGQYTNKELRKXXEDGMRDNPM) have homology to various complement C3a components. Listericidin also kills other Listeria spp., Vibrio spp., Salmonella spp., Escherichia spp., Cronobacter spp., and Bacillus spp. The listericidin peptide purified in a single-step chromatography is pH and heat stable, and has a broad antimicrobial spectrum against major foodborne pathogens in addition to L. monocytogenes.
authors.affiliation: Division of Virulence Assessment, Food and Drug Administration , Laurel, Maryland.
authors.forename: Mahendra H
authors.initials: MH
authors.lastname: Kothary
authors.affiliation: Division of Virulence Assessment, Food and Drug Administration , Laurel, Maryland.
authors.forename: Augusto A
authors.initials: AA
authors.lastname: Franco
authors.affiliation: Division of Virulence Assessment, Food and Drug Administration , Laurel, Maryland.
authors.forename: Ben D
authors.initials: BD
authors.lastname: Tall
authors.affiliation: Division of Virulence Assessment, Food and Drug Administration , Laurel, Maryland.
authors.forename: Gopal R
authors.initials: GR
authors.lastname: Gopinath
authors.affiliation: Division of Virulence Assessment, Food and Drug Administration , Laurel, Maryland.
authors.forename: Atin R
authors.initials: AR
authors.lastname: Datta
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000097:blood* / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001798:Blood Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D015926:Complement C3a / Q000737:chemistry / Q000276:immunology; D005516:Food Microbiology; D005517:Foodborne Diseases / Q000382:microbiology; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008089:Listeria monocytogenes / Q000187:drug effects*; D011817:Rabbits / Q000097:blood*; D020539:Sequence Analysis, Protein; D017385:Sequence Homology; D012965:Sodium Chloride / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D001798:Blood Proteins; C031817:listericidin; D012965:Sodium Chloride; D015926:Complement C3a