eprintid: 10191358 rev_number: 12 eprint_status: archive userid: 699 dir: disk0/10/19/13/58 datestamp: 2024-06-06 08:12:08 lastmod: 2024-06-06 08:12:08 status_changed: 2024-06-06 08:12:08 type: thesis metadata_visibility: show sword_depositor: 699 creators_name: Webley, Catherine Ruth title: Pore-Forming Behaviour of Nisin and Nisin-Mimetics in Model Membranes ispublished: unpub divisions: UCL divisions: B04 divisions: C06 divisions: F56 note: Copyright © The Author 2022. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. abstract: Antimicrobial resistance is a global health threat that requires innovative solutions. The antimicrobial peptide nisin is promising due to its non-toxicity and low resistance rates despite widespread use as a food preservative. However, nisin has poor oral bioavailability, solubility, and stability. Nisin has a dual mode of action. This involves binding and sequestration of Gram-positive bacterial cell wall precursor lipid II. Additionally, nisin forms nanopores that puncture the bacterial cell membrane causing efflux of cell nutrients and collapse of the cell ionic potential. The dominant mode of action and precise nanopore architecture are currently unknown. In this work, biophysical techniques were applied to unresolved questions concerning nisin’s pore-forming activity. Single-channel current recordings were used to determine nisin’s pore conductance and pore size. Measurements on model membranes with variable anionic content revealed how the lipid environment influences peptide-mediated poration. Larger pores of ∼ 2 nm diameter were observed in membranes with higher anionic content. This may reflect nisin activity in biological systems. Native nisin was compared with semi-synthetic structural analogues to probe linearisation of up to three of nisin’s five thioether bridged lanthionine rings. Antimicrobial activity of the bacterial cell membrane-targeting nisin AB rings conjugated onto a linear pore-forming peptide was investigated alongside the role of nisin’s C ring (the hinge region). Findings suggest retention of pore-forming activity following linearisation of nisin’s C, D and E rings with a reduction in pore diameter. Insight gained may inform design of nisin-like antibiotics with improved drug properties and unique modes of action. date: 2024-04-28 date_type: published full_text_type: other thesis_class: doctoral_embargoed thesis_award: Ph.D language: eng verified: verified_manual elements_id: 2270699 lyricists_name: Webley, Catherine Ruth lyricists_id: CRWEB44 actors_name: Webley, Catherine actors_id: CRWEB44 actors_role: owner full_text_status: restricted pagerange: 1-233 pages: 233 institution: UCL (University College London) department: Chemistry thesis_type: Doctoral editors_name: Howorka, Stefan editors_name: Tabor, Alethea citation: Webley, Catherine Ruth; (2024) Pore-Forming Behaviour of Nisin and Nisin-Mimetics in Model Membranes. Doctoral thesis (Ph.D), UCL (University College London). document_url: https://discovery.ucl.ac.uk/id/eprint/10191358/2/Webley_19100571_PhD_Thesis.pdf