Studies of the highly potent lantibiotic peptide nisin Z in aqueous solutions of salts and biological buffer components

Abstract

The lantibiotic nisin, usually used as a 2.5%w/w in NaCl and milk solids, has activity against a wide range of Gram-positive bacteria, especially food-borne pathogens, and has been used as a food preservative for decades without the development of significant resistance. It has been reported that the high purity (>95%) nisin Z form has activity against the Gram-negative speciesE. coli, which is significantly reduced in the presence of NaCl. This current study examined, by1H NMR spectroscopy, the effects of NaCl, and a range of other salts, on the observed aqueous solution1H NMR spectra of nisin Z in the pH 3–4 range, where nisin Z has its maximum stability. Nisin's mechanism of action involves binding to the polyoxygenated pyrophosphate moiety of lipid II, and in acidic solution the positively charged C-terminus region is reported to interact with the negative sulfate groups of SDS micelles, so the study was extended to include a number of polyoxygenated anions commonly used as buffers in many biological assays. In general, the biggest changes found were in the chemical shifts of protons in the hydrophobic N-terminus region, rather than the more polar C-terminus region. The effects seen on the addition of the salts (cations and anions) were not just an overall non-specific ionic strength effect, as different salts caused different effects, in an unpredictive manner. Similarly, the polyoxygenated anions behaved differently and not predictably, and neither the cations/anions, or polyoxygenated anions, constitute a Hofmeister or inverse Hofmeister series.