title: Isolation, biochemical characterization, and cloning of a bacteriocin from the poultry-associated Staphylococcus aureus strain CH-91.
abstract: Staphylococcus aureus strain CH-91, isolated from a broiler chicken with atopic dermatitis, has a highly proteolytic phenotype that is correlated with the disease. We describe the isolation and biochemical and molecular characterization of the AI-type lantibiotic BacCH91 from S. aureus CH-91 culture medium. The bacteriocin was purified using a three-stage procedure comprising precipitation with ammonium sulfate, extraction with organic solvents, and reversed-phase HPLC. The BacCH91 peptide is thermostable and highly resistant to cleavage by both prokaryotic and eukaryotic peptidases. The MIC for the Gram-positive bacteria ranged from 2.5 nM for Microococcus luteus through 1.3-6.0 μM for staphylococcal strains up to more than 100 μM for Lactococcus lactis. BacCH91 was ineffective against the Gram-negative strains tested at the maximal concentration (100 μM). The amino acid sequence of BacCH91 is similar to that of epidermin and gallidermin. The encoding gene (bacCH91) occurred in two allelic variants distinguishable in the restriction fragment length polymorphism assay. Variant I, identified in S. aureus CH-91, dominated in S. aureus strains of poultry origin, although strains with variant II were also identified in this group. S. aureus strains of human origin were characterized exclusively by variant II.
authors.affiliation: Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland.
authors.forename: Benedykt
authors.initials: B
authors.lastname: Wladyka
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Wielebska
authors.forename: Marcin
authors.initials: M
authors.lastname: Wloka
authors.forename: Oliwia
authors.initials: O
authors.lastname: Bochenska
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Dubin
authors.forename: Adam
authors.initials: A
authors.lastname: Dubin
authors.forename: Pawel
authors.initials: P
authors.lastname: Mak
mesh_terms: D000818:Animals; D001430:Bacteriocins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D005591:Chemical Fractionation; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D013294:Lactococcus lactis / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D016982:Micrococcus luteus / Q000187:drug effects; D008969:Molecular Sequence Data; D011200:Poultry; D011201:Poultry Diseases / Q000382:microbiology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013203:Staphylococcal Infections / Q000382:microbiology / Q000662:veterinary; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D001430:Bacteriocins; D004269:DNA, Bacterial