Staphylococcins: an update on antimicrobial peptides produced by staphylococci and their diverse potential applications.

Staphylococcins are antimicrobial peptides or proteins produced by staphylococci. They can be separated into different classes, depending on their amino acid composition, structural complexity, and steps involved in their production. In this review, an overview of the current knowledge on staphylococcins will be presented with emphasis on the information collected in the last decade, including a brief description of new peptides. Most staphylococcins characterized to date are either lantibiotics or linear class II bacteriocins. Recently, gene clusters coding for production of circular bacteriocins, sactipeptides, and thiopeptides have been mined from the genome of staphylococcal isolates. In contrast to class II bacteriocins, lantibiotics, sactipeptides, and thiopeptides undergo post-translational modifications that can be quite extensive, depending on the peptide. Few staphylococcins inhibit only some staphylococcal species, but most of them have proven to target pathogens belonging to different genera and involved in a variety of infectious diseases of clinical or agronomic importance. Therefore, these peptides exhibit potential application as anti-infective drugs in different areas. This review will also cover this diverse and remarkable potential. To be commercialized, however, staphylococcin production should be cost-effective and result in high bacteriocin yields, which are not generally achieved from the culture supernatant of their native producers. Such low yields make their production quite costly and not suitable at large industrial scale. Efforts already made to overcome this limitation, minimizing costs and time of production of some staphylococcins and employing either chemical synthesis or in vivo biosynthesis, will be addressed in this review as well. KEY POINTS: • Staphylococci produce a variety of antimicrobial peptides known as staphylococcins. • Most staphylococcins are post-translationally modified peptides. • Staphylococcins exhibit potential biotechnological applications. Graphical abstract.

Molecular characterization of aureocin 4181: a natural N-formylated aureocin A70 variant with a broad spectrum of activity.

Bacteriocins are ribosomally synthesized antimicrobial peptides produced by prokaryotes. Here, the molecular characterization of aureocin 4181, a bacteriocin produced by Staphylococcus aureus 4181, a strain involved in bovine mastitis, is presented. Aureocin 4181 gene cluster (aurRID1CBAT) was mined from scaffold 15 of the draft genome of its producer strain. Three (AurABC) out of the four structural peptides of aureocin 4181 are identical to those of aureocin A70, except for AurD1 of aureocin 4181, which showed a conservative substitution of Leu29 to Phe29 when compared to AurD of aureocin A70. According to molecular mass determination and peptide sequencing, combined with genome sequencing data, aureocin 4181 is an N-formylated variant of aureocin A70. The analysis of its antimicrobial spectrum was extended to include strains of the two major contagious pathogens involved in bovine mastitis, S. aureus and Streptococcus agalactiae. Aureocin 4181 exhibited a striking activity against S. aureus, inhibiting most strains tested. Besides having a broader spectrum of activity, aureocin 4181 exhibited a stronger bacteriolytic action against the target strains and proved to be from two- to fourfold more active than aureocin A70 against S. aureus. Aureocin 4181 has potential to become an alternative drug for prevention and control of mastitic staphylococci, a pathogen that imposes a huge economic burden to dairy industry worldwide. It also represents the third four-component bacteriocin described in the literature, the second in staphylococci.

Genetic and biochemical characterization of hyicin 3682, the first bacteriocin reported for Staphylococcus hyicus.

Hyicin 3682, the first bacteriocin reported for Staphylococcus hyicus, is a BsaCOL variant produced by S. hyicus 3682, a strain isolated from bovine milk. Hyicin 3682 is found in the culture supernatant, is bactericidal and its producing strain exhibits a much broader spectrum of antimicrobial activity than the producing strain of BsaCOL against several Gram-positive bacteria, which include foodborne pathogens, food-spoilage microorganisms and bacterial species of medical and veterinary importance. Sequencing of the genome of S. hyicus 3682 provided the nucleotide sequence of the entire gene cluster involved in hyicin 3682 production, which seems to be located on pRJ109, the single plasmid carried by this strain. This gene cluster is expressed and consists of 8525bp and of eight genes (hyiA, hyiB, hyiC, hyiD, hyiP, hyiF, hyiE and hyiG) encoded on the same DNA strand. The mature lantibiotic exhibits 91% identity to BsaCOL and its molecular mass was found to be ∼26Da higher due to two amino acid substitutions. S. hyicus 3682 proved to be only partially immune to its cognate bacteriocin up to 1024 AU/ml. Therefore, hyicin 3682, the first Bsa variant reported in coagulase-negative staphylococci, does exhibit antimicrobial and siblicidal activities.

The four-component aureocin A70 as a promising agent for food biopreservation.

Aureocin A70 is the only four-component bacteriocin described to date. As it inhibits the growth of a wide range of Gram-positive bacteria, including Listeria monocytogenes strains isolated from food, its potential for improving food safety was investigated in this study. Aureocin A70 (10,240AU/mL) proved to be bactericidal, but not extensively lytic, against listerial strains. The antibacterial activity of aureocin A70 (16AU/mL) was then tested in UHT-treated skimmed milk inoculated with the food-associated L. monocytogenes L12 strain (4-log CFU/mL) during storage at 4°C for one week. Aureocin A70 caused a time-dependent reduction in the listerial viable cell counts (5.51-log units) up to 7days of incubation. Aureocin A70 was neither toxic to the Vero and the L-929 cell lines nor exhibited a hemolytic activity against sheep red blood cells. Aureocin A70 proved to be completely stable for one month at 25°C, 16weeks at 4°C and 20weeks at -20°C. Aureocin A70 exhibited a time-dependent susceptibility to simulated gastric juice and bile salts mimicking gastrointestinal conditions. The entrapment of aureocin A70 in an alginate/gelatin matrix revealed that this bacteriocin can be released from this matrix. Moreover, it remained adsorbed to and active on a low-density polyethylene plastic surface suggesting that aureocin A70 may be employed in bioactive packaging to control the growth of undesirable bacteria. Taken together these results suggest that aureocin A70 is a promising alternative to be used in food applications.