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Analysis and exploitation of AHFCA-dependent signalling systems in Streptomyces bacteria

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Poon, Vincent (2015) Analysis and exploitation of AHFCA-dependent signalling systems in Streptomyces bacteria. PhD thesis, University of Warwick.

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Official URL: http://webcat.warwick.ac.uk/record=b2870443~S1

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Abstract

From genome sequencing and analyses, Streptomyces bacteria are predicted to produce hundreds of bioactive metabolite compounds; however, the products of some of these cryptic biosynthetic gene clusters are most often silent or produced in very small quantities in laboratory conditions. The AHFCA-dependent signalling system is an example of a regulatory systems that tightly control expression of cryptic biosynthetic gene clusters in Streptomyces bacteria. Bioinformatic analyses, using MEME and FIMO, were able to successfully reveal TetR repressor binding sites to gain insights into complex regulatory networks.

The S. avermitilis cryptic biosynthetic gene cluster, which is proposed to be under the control of an AHFCA-dependent signalling system, has not been investigated before and genome mining predictions suggest the biosynthetic gene cluster could be involved in the biosynthesis of novel azoxy compounds. The 50 kb predicted azoxy biosynthetic gene cluster was captured using pESAC13A PAC technologies and introduced into S. coelicolor M1152 and S. lividans TK24 heterologous hosts. Comparative metabolic profiling successfully identified the predicted azoxy compounds in S. coelicolor M1152 by LC-MS and identification of the azoxy compounds were supported by UHPLC-ESI-TOF-MS analysis and incorporation of isotope-labelled L-serine.

Consistent with the model of transcriptional regulation, exploitation of the S. avermitilis AHFCA-dependent signalling system through the inactivation of sav_2268 transcriptional repressor in S. lividans TK24 heterologous hosts unlocked the production of azoxy compounds and AHFCA signalling molecules.

Item Type: Thesis (PhD)
Subjects: Q Science > QR Microbiology
Library of Congress Subject Headings (LCSH): Streptomyces, Cellular signal transduction
Official Date: December 2015
Dates:
DateEvent
December 2015Submitted
Institution: University of Warwick
Theses Department: Systems Biology Doctoral Training Centre
Thesis Type: PhD
Publication Status: Unpublished
Supervisor(s)/Advisor: Corre, Christophe ; Moore, Jonathan D.
Sponsors: University of Warwick. Systems Biology Doctoral Training Centre ; Biotechnology and Biological Sciences Research Council (Great Britain)
Extent: xviii, 221 leaves : illustrations, charts
Language: eng

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