Characterisation of a bacterial P450 system using surface mass spectrometry

  • Andrés Galindo García

Student thesis: Master of Philosophy

Abstract

In this study, the components of a biotechnologically relevant E. coli expressionsystem were characterised through the use of mass spectrometry techniques. Theseinclude the bacterial host itself, the enzymatic product of the P450 RhF geneexpression which catalyses a selective hydroxylation reaction that produces 5-hydroxydiclofenac from a diclofenac substrate; and both the substrate and productof the reaction. The identification of molecular signals as phosphatidylglicerol (PG),phosphatidylethanolamine (PE) and fatty acids (FA) led to the generation of a lipidprofile. Instrumentation relying on different ionisation mechanisms was exploredand bioinformatic tools were used to extract useful information from large imagingdata sets through normalisation and PCA analysis.
Date of Award3 Jan 2017
Original languageEnglish
Awarding Institution
  • The University of Manchester
SupervisorNicholas Lockyer (Supervisor) & Sabine Flitsch (Supervisor)

Keywords

  • P450
  • Bacterial analysis
  • Biotechnology
  • DESI
  • REIMS
  • Enzymatic system
  • Mass spectrometry
  • SIMS
  • Lipid profile

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