Objective
THE AIM OF THIS PROJECT IS TO IMPROVE AND TO DEVELOP GENETICALLY ENGINEERED AND/OR MUTANT YEAST STRAINS WHICH OVERPRODUCE AND EXCRETE AMINO-ACIDS AND TO DETERMINE CONTINUOUS CULTURE CONDITIONS UNDER WHICH THE POTENTIAL OF SUCH STRAINS CAN BE COMPETITIVELY EXPLOITED.
A biochemical and genetic study was made of the mechanisms involved in amino acid overproduction and export in the yeast Saccharomyces cerevisiae, using aromatic amino acids as the experimental system. The metabolic bottlenecks opposing maximum product formation in deregulated and/or transformed strains were investigated. Mass culture conditions were analysed and a partially automatic laboratory fermentation system was developed ensuring the most efficient product formation by the developed strains.
Mutations favouring overproduction and excretion of aromatic amino acids were isolated. Strains bearing 1 or several of these mutations were constructed and their enzymatic, growth and excretory properties examined. Certain implicated genes were cloned and sequenced, making it possible to manipulate them in vitro. This is the case of the prephenate dehydrogenase gene, which was shown to play a decisive role in channelling the metabolic flow into tyrosine or phenylalanine production.
A sophisticated automated fermentation control system, adaptable to genetically modified yeast strains, was developed and is being improved constantly. The studies concerning growth inhibition of overproducing strains revealed the role of excess acetate formation and the dysfunction of the aromatic transamination reactions.
THIS RESEARCH PROJECT WILL BE FOCUSED ON THE DEVELOPMENT OF YEAST STRAINS WHICH OVERPRODUCE AND EXCRETE AMINO ACIDS. THE METHODOLOGY WILL INVOLVE CLASSICAL AND DIRECTED MUTAGENESIS, AND BY COMBINATIONS OF PROMISING MUTATIONS BY CLASSICAL GENETIC METHODS OR BY GENE CLONING INTO HIGH EXPRESSION PLASMID VECTORS. CULTURE TECHNIQUES WILL INCLUDE CLASSICAL CULTIVATION IN FLASKS AND IN 20 L FERMENTERS, WITH EMPHASIS ON CONTINUOUS CULTURES, AS WELL AS THE EMERGING TECHNIQUES OF ALL RECYCLING AND IMMOBILIZATION.
THE STRATEGIES WILL BE :
1. ELIMINATION OF METABOLIC BOTTLENECKS
2. ENHANCEMENT OF AMINO-ACID EXPORT
3. FINDING OPTIMAL GROWTH CONDITIONS FOR HIGH PRODUCTION
4. DEVELOPMENT OF A HIGHLY AUTOMATIC FERMENTATION CONTROL SYSTEM.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- engineering and technology electrical engineering, electronic engineering, information engineering electronic engineering control systems
- natural sciences biological sciences genetics mutation
- natural sciences chemical sciences organic chemistry amines
- engineering and technology industrial biotechnology bioprocessing technologies fermentation
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Coordinator
1050 BRUSSELS
Belgium
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