Award details

Analysis of the dynamics and robustness of metabolic networks in a genetically engineered Pseudomonas fluorescens with inducible exo-polysaccharide pr

Reference	BB/F003447/1
Principal Investigator / Supervisor	Professor Royston Goodacre
Co-Investigators / Co-Supervisors
Institution	The University of Manchester
Department	Chemistry
Funding type	Research
Value (£)	290,346
Status	Completed
Type	Research Grant
Start date	01/10/2007
End date	30/09/2010
Duration	36 months

Abstract

By a Systems Biology approach to explore the dynamics and robustness of metabolic networks in genetically engineered Pseudomonas fluorescens bacteria in which the level of production of the industrially important and medically relevant exo-polysaccharide alginate can be controlled over a wide range. Targeted and random gene interruption mutants affected in alginate synthesis and mutants expressing modified levels of key enzymes will also be included in the studies. The alginate production will be controlled by addition of an externally added inducer, and at maximum levels of induction more than 30% of the total sugar consumption will be channelled into alginate production. To reduce interpretation complexity the bacteria will be studied in continuous cultivations at steady-state conditions and through time-course induction experiments. The cellular responses to induction of alginate synthesis will be analyzed at the RNA, protein and metabolite levels, including metabolic flux analysis. Targeted mutations and a global screen of a transposon insertion library will be used to identify genes not known to affect alginate synthesis. A 'Top-down' strategy using novel evolutionary computing will be linked to a 'Bottom-up' strategy with construction of a genome-scale metabolic model for integrative 'omics analysis and whole cell simulations.

Summary

Pseudomonas fluorescens is a bacterium which can be made to produce the industrially important and medically relevant exo-polysaccharide alginate. Aliginate is a type of gum which is generally obtained from brown algae. It is used by the food industry to increase viscosity and as an emulsifier of their products, and it is also used in medicine in indigestion tablets. Extracting alginate from brown algae is a complex process and a cleaner method is to make bacteria excrete this polysaccharide. The alginate can then be easily recovered from fermentors. However, the yield of alginate production in Pseudomonas fluorescens is generally low and we hope to increase the production of this valuable product by understanding the biological system more fully. In this project, we will develop a Systems Biology approach to explore the dynamics and robustness of metabolic processes in genetically engineered Pseudomonas fluorescens bacteria in which the level of alginate production can be controlled over a wide range. Targeted and random gene interruption mutants affected in alginate synthesis and mutants expressing modified levels of key enzymes will also be included in the studies and this shall help us increase our knwoledge of this complex biological process.

Committee	Closed Committee - Engineering & Biological Systems (EBS)
Research Topics	Industrial Biotechnology, Microbiology, Synthetic Biology, Systems Biology
Research Priority	X – Research Priority information not available
Research Initiative	Systems Biology of Microorganisms (SysMo) [2007-2008]
Funding Scheme	X – not Funded via a specific Funding Scheme

I accept the terms and conditions of use (opens in new window)

export PDF file

back to list new search