![[BioMatNet Database - FP5 Quality of Life Programme]](../images/FP5.gif) |
QLK3-2000-00497
Construction of Corynebacterium glutamicum strains producing either L-valine or D-pantothenic acid - A rational approach using genome research |
| Contract No: | QLK3-2000-00497 |
| Project Type: | RS (Research and Technological Development Project) |
| Start Date: | 01-02-2001 |
| Duration: | 36 months |
| Total Cost: | |
| EC Contribution: | 2 085 175 EUR |
| Scientific Officer: |
Abstract
Amino acids and vitamins play an essential role in economic and ecologically- sociable animal nutrition in stock farming. Corynebacterium glutamicum has been chosen as a bacterial cell factory to study the biological production of the amino acid L-valine and the vitamin D-pantothenic acid by a rational approach using genome research for metabolic engineering. This approach is based on a whole-genome analysis with the principle idea to use genome-wide data and innovative technologies such as DNA chip technology, transcriptomics and proteomics. Genetic and metabolic targets will be identified in multi-disciplinary approaches to develop new generations of production strains. Small-scale simulation studies will provide a sound basis for the subsequent industrial scale-up.
Objectives
The principle problem to be addressed by the VALPAN consortium is to achieve an optimal use of feed additives in stock farming with amino acids and vitamins produced by an ecologically clean fermentation process. For this purpose, Corynebacterium glutamicum, an amino acid producing bacterial cell factory has been chosen for a genome-wide rational approach to overproduce the amino acid L-valine and the vitamin D-pantothenic acid. The impact of this research will be on reducing the use of chemically synthesised amino acids and vitamins in feed additives. New generations of production strains better able to rapidly produce a high concentration of the new products will be developed. The data obtained will be used to improve current fermentation strategies.
Description of the work
The VALPAN project comprises six work packages (WP):
WP1: Will analyse the regulation of the biosynthesis of valine and pantothenic acid and establish controlled-expression systems using inducible derepressible promoters. The valine stress response will be investigated. Using the combined knowledge of the project new generations of production strains will be constructed.
WP2: Will characterise the precursor supply for the biosynthesis of valine and pantothenate. Enzymes involved in biosynthesis will be characterised biochemically. The effect of changes in cellular physiology and biochemistry on valine biosynthesis will be studied.
WP3: Will use whole-cell kinetic analyses and physiological data of intracellular pathway fluxes for flux modelling of production strains. Uptake and excretion systems will be identified and characterised.
WP4: Will establish DNA chip technology for global expression analyses of C. glutamicum.during valine and pantothenate production. Bioinformatic tools will allow the identification of new targets for the genetic improvement of production strains.
WP5: Will perform physiological studies on the scale-up/scale-down of C. glutamicum.fermentation for the production of valine and pantothenic acid.
WP6: Will co-ordinate the whole VALPAN project and will manage the whole-genome database of C. glutamicum.
Milestones
Genetic analysis of all genes necessary for valine and pantothenate production and construction of specific tools for controlled gene expression will result in new generations of production strains. Biochemical, kinetic and regulatory data on purified enzymes involved in precursor supply for valine and pantothenate synthesis will allow the optimisation of precursor supply in production strains. Recombinant strains with altered activities of enzymes involved in precursor supply will be constructed.
Deliverables
Carbon flux analysis for valine and pantothenate overproducing strains and data on the optimal energy state for production will be used to develop a metabolic flux model for production strains. Altered uptake and/or excretion systems for both products will be used for the construction of production strains. DNA chips representing all genes of the C. glutamicum genome and bioinformatics databases for input, retrieval and analysis of genome expression data will be established. This is the prerequisite for a global expression analysis of C. glutamicum strains during valine and pantothenate production. Data on bacterial population dynamics and single cell properties in response to a fluctuating microenvironment in small-scale simulations and large-scale fermentation trials will allow a subsequent scale-up of C. glutamicum fermentations.
Contacts
Coordinator
Participant
© Copyright 2006 Policy Statements
Updated
by CPL Press:
03/07/2007
- biomatnet@biomatnet.org
 |
 |
 |
News |
 |
Events |
|
|