RESUMO
INTRODUCTION: Honey is known for exhibiting antibacterial properties, indicating its use as part of traditional medicine since the early ages. With the advent of antibiotic-resistant bacteria, the need for alternative antimicrobials has outpaced the actual development of novel, broad-spectrum antibiotics. Previous research has revolved around the sugar content of honey because its sweetness makes it an attractive food source. However, research assessing the protein and lipid components of honey is lagging behind that of its sugar counterpart. The goal of this investigation was to examine the antimicrobial properties of honey and to identify any distinct proteins or lipids. METHODS: In order to isolate individual peptides and lipids, the different samples of local and foreign-sourced honeys were dialyzed, and the resulting dialysate proteins were screened via gel electrophoresis (sodium dodecyl-sulfate polyacrylamide gel electrophoresis [SDS-PAGE]) with Coomassie blue and silver stain, while lipids were examined using thin layer chromatography (TLC). To assess antimicrobial potency, a series of Kirby-Bauer disc diffusion assays was performed on Mueller-Hinton agar using different types of raw honey with Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia coli, and Bacillus subtilis. The process was then repeated using the peptide extracts from the dialyzed fractions of the honeys. RESULTS: The SDS-PAGE trials revealed repetitive promising protein bands across several gels below 75kDa with both Coomassie blue and silver staining. The TLC analysis of varying raw honey samples consistently demonstrated the presence of medium and long-chain fatty acids, likely in the range of C12-C14. In the disc diffusion assays, the greatest amount of inhibition was seen when the honeys were tested as a whole instead of its constituent parts. CONCLUSION: Instead of an individual component acting as the key to honey's action against bacteria, it appears there is a synergistic relationship amongst the sugars, proteins, and lipids that make each honey unique.
RESUMO
In enterohemorrhagic Escherichia coli (EHEC), sigma factor N (σ(N)) regulates glutamate-dependent acid resistance (GDAR) and the locus of enterocyte effacement (LEE); discrete genetic systems that are required for transmission and virulence of this intestinal pathogen. Regulation of these systems requires nitrogen regulatory protein C, NtrC, and is a consequence of NtrC-σ(N) -dependent reduction in the activity of sigma factor S (σ(S)). This study elucidates pathway components and stimuli for σ(N)-directed regulation of GDAR and the LEE in EHEC. Deletion of fliZ, the product of which reduces σ(S) activity, phenocopied rpoN (σ(N)) and ntrC null strains for GDAR and LEE control, acid resistance, and adherence. Upregulation of fliZ by NtrC-σ(N) was shown to be indirect and required an intact flagellar regulator flhDC. Activation of flhDC by NtrC-σ(N) and FlhDC-dependent regulation of GDAR and the LEE was dependent on σ(N)-promoter flhDP 2 , and a newly described NtrC upstream activator sequence. Addition of ammonium chloride significantly altered expression of GDAR and LEE, acid resistance, and adherence, independently of rpoN, ntrC, and the NtrC sensor kinase, ntrB. Altering the availability of NtrC phosphodonor acetyl phosphate by growth without glucose, with acetate addition, or by deletion of acetate kinase ackA, abrogated NtrC-σ(N)-dependent control of flhDC, fliZ, GDAR, and the LEE.