BHT-B

Streptococcus rattus strain BHT is a species representative and strong bacteriocin producer. Here we report that S. rattus BHT produces two quite different types of bacteriocin activity, named BHT-A and BHT-B. The two bacteriocins were purified and analysed for activity and by MALDI-TOF mass spectrophotometry. BHT-A was found to be a variant of the two-component lantibiotic, Smb. BHT-B is a non-modified 5195Da peptide with some similarity to the tryptophan-rich Staphylococcus aureus bacteriocin, aureocin A53. Six S. rattus and two S. mutans strains were found to contain both the BHT-A and BHT-B genetic loci.

Sodium lactate and acetic acid derivatives were evaluated for their impact on fresh Italian pork sausage using commercial trimmings. Analysis over storage included total plate count (TPC), TBARS, sensory analysis, cooking loss, pH, and color. Treatments included: (a) vinegar and sodium lactate mixture (V), (b) sodium lactate (S), (c) positive control with BHA/BHT (B) and (d) negative control, seasoning only (C). Treatments S and V had lower TPC (P0.05) over time. While CIE a* surface values for redness generally decreased over storage time for all treatments, B maintained more redness. There were few major differences in descriptive sensory evaluation over time, but S and V precluded early onset of rancidity, oxidation and other off-flavors contrary to some of the analytical results. Of consumers tested, 85.6% rated all treatments between like slightly and like very much.

The emergence of antibiotic-resistant bacteria led to an urgent need for next-generation antimicrobial agents with novel mechanisms of action. The use of positively charged antimicrobial peptides that target cytoplasmic membrane is an especially promising strategy since essential functions and the conserved structure of the membrane hinder the development of bacterial resistance. Aureocin A53- and enterocin L50-like bacteriocins are highly cationic, membrane-targeting antimicrobial peptides that have potential as next-generation antibiotics. However, the mechanisms of resistance to these bacteriocins and cross-resistance against antibiotics must be examined before application to ensure their safe use. Here, in the model bacterium Lactococcus lactis, we studied the development of resistance to selected aureocin A53- and enterocin L50-like bacteriocins and its correlation with antibiotics. First, to generate spontaneous resistant mutants, L. lactis was exposed to bacteriocin BHT-B. Sequencing of their genomes revealed single nucleotide polymorphisms (SNPs) in the dgkB (yecE) and dxsA genes encoding diacylglycerol kinase and 1-deoxy-D-xylulose 5-phosphate synthase, respectively. Then, selected mutants underwent susceptibility tests with a wide array of bacteriocins and antibiotics. The highest alterations in the sensitivity of studied mutants were seen in the presence of cytoplasmic membrane targeting bacteriocins (K411, Ent7, EntL50, WelM, SalC, nisin) and antibiotics (daptomycin and gramicidin) as well as lipid II cycle-blocking bacteriocins (nisin and Lcn972) and antibiotics (bacitracin). Interestingly, decreased via the SNPs accumulation sensitivity to membrane-active bacteriocins and antibiotics resulted in the concurrently increased vulnerability to bacitracin, carbenicillin, or chlortetracycline. It is suspected that SNPs may result in alterations to the efficiency of the nascent enzymes rather than a total loss of their function as neither deletion nor overexpression of dxsA restored the phenotype observed in spontaneous mutants.