Svatoš, Alma (2019) Structural and biochemical characterization of carrier proteins interacting with non-elongating ketosynthase from enacyloxin polyketide synthase. PhD thesis, University of Warwick.

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Abstract

Enacyloxin is an antibiotic synthesized inside bacterial cells by gigantic modular multienzyme complex called polyketide synthase (PKS), which is active against multidrug-resistant Acinetobacter baumannii that leads the World Health Organization ‘priority pathogens’ list. In spite of its promising physiological activity, enacyloxin IIa has not been used as a drug due its structural instability. The modular nature of enacyloxin and other PKSs makes them amenable to rational engineering to elucidating protein-protein interactions within PKSs will be crucial for their bioengineering to produce novel drugs. A key step in enacyloxin biosynthesis is the transfer of the polyketide chain from an acyl carrier protein (ACP) domain in the last PKS module to a standalone peptidyl carrier protein (PCP) by a non-elongating ketosynthase (KS^0) domain. It has been reported that this transfer is necessary for the condensation (C) domain to catalyse polyketide chain release via intermolecular esterification with 3, 4-dihydroxycyclo-hexane carboxylic acid (DHCCA). No esterification occurs when the substrate is attached to the ACP domain. Thus, the KS^0 transfers the substrate from a carrier protein that is unable to interact with the C domain to one that it is able to interact with.

An experimental approach combining NMR and biochemical techniques presented in the thesis suggests that the length of helix 1 and the lack of charged residues in the C-terminal part of ACP prevent it from establishing favourable interaction with C domain.

Item Type:	Thesis (PhD)
Subjects:	Q Science > QD Chemistry Q Science > QP Physiology R Medicine > RM Therapeutics. Pharmacology
Library of Congress Subject Headings (LCSH):	Antibiotics -- Development, Antibiotics -- Synthesis, Carrier proteins, Polyketides, Biosynthesis, Natural products -- Biotechnology
Official Date:	30 September 2019
Dates:	Date Event 30 September 2019 UNSPECIFIED
Institution:	University of Warwick
Theses Department:	Department of Chemistry
Thesis Type:	PhD
Publication Status:	Unpublished
Supervisor(s)/Advisor:	Lewandowski, Józef R. ; Challis, Gregory L.
Sponsors:	European Research Council
Format of File:	pdf
Extent:	viii, 160 leaves : illustrations (some colour)
Language:	eng

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