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Characterisation of a bacterial P450 system using surface mass spectrometry
[Thesis]. Manchester, UK: The University of Manchester; 2016.
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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.
Keyword(s)
Bacterial analysis; Biotechnology; DESI; Enzymatic system; Lipid profile; Mass spectrometry; P450; REIMS; SIMS