Thesis

Investigating the response of pseudomonas aeruginosa PA14 to acrylic monomers and cystic fibrosis therapeutics for industrial and medical application

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Awarding institution
  • University of Strathclyde
Date of award
  • 2024
Thesis identifier
  • T16897
Person Identifier (Local)
  • 201956645
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Department, School or Faculty
Abstract
  • Increasing global demand for high-clarity and high tensile strength plastic has led to industrial interest in a fermentation process for monomer production from non-petrochemical sources. The main roadblock in a monomer fermentation is the toxicity of the solvent product to the production organism. Pseudomonas spp. are promising candidates as production organisms as their intrinsic levels of solvent tolerance is well known. Intrinsic levels of tolerance to many substances allows Pseudomonas aeruginosa to persist in infection, particularly demonstrated during infection in patients suffering with cystic fibrosis. PA14 transposon insertion mutants of the genes encoding subunits of the MexAB-OprM multi-drug efflux system show increased sensitivity to industrial solvents compared to the wild type. We have identified a group of membrane bound metalloproteins belonging to the Cbb3 cytochrome oxidase family and an associated maturation system which also show an increase in solvent sensitivity after transposition. We believe this family of metalloproteins drives RND efflux activity by maintaining a proton gradient within the cell. We demonstrate the purification of subunits of the MexAB-OprM system and Cbb3 proteins belonging to the maturation system that we believe are essential for solvent tolerance in PA14 for functional characterization using solid supported membrane electrophysiology. The transcriptomic response of P. aeruginosa PA14 to the industrial solvents BMA, styrene and ethylbenzene alongside the cystic fibrosis transmembrane conductance regulator modulator drugs Kaftrio and Ivacaftor is described. We provide further evidence of a functional relationship between resistance nodulation division type efflux systems and the Cbb3 cytochrome oxidases and suggest its application in bioprocess development. A transcriptional response to Kaftrio and Ivacaftor is also described, providing the basis for further study on the effect of these therapeutics on P. aeruginosa in vivo. We therefore demonstrate biochemical, transcriptomic and genetic approaches for the detection of tolerance conferring systems in P. aeruginosa PA14 for industrial and medical benefit.
Advisor / supervisor
  • Javelle, Arnaud
  • Tucker, Nicholas
Resource Type
Note
  • Previously held under moratorium from 15 March 2024 until 15 March 2026.
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