Objective
AN INVESTIGATION OF THE BEHAVIOUR OF ORGANISMS IN CHEMOSTAT CULTURE SHOULD ALLOW ASSESSMENTS TO BE MADE OF METABOLIC ACTIVITY RELATIVE TO GROWTH IN PRESCRIBED ENVIRONMENTS AND THEREBY ALLOW METABOLITE FLOW TO BE STEERED FROM CENTRAL PATHWAYS OF METABOLISM TO INDUSTRIALLY IMPORTANT PRODUCTS. ONCE THE CONTROLS HAVE BEEN INDENTIFIED, THE FACTORS REQUIRED TO OPTIMIZE THE PRODUCTION OF A METABOLITE MAY BE UNDERSTOOD, AND A STRATEGY ADOPTED FOR THE DEVELOPMENT OF AN INDUSTRIAL PROCESS.
Novel thermotolerant methylotrophic bacilli were isolated in order to elucidate their pathways(s) of methanol metabolism and mode(s) of regulation. By means of genetic and physiological manipulation, metabolism was guided towards an overproduction of important biochemicals. Environmental parameters were independently manipulated using chemostat culture techniques and specific gene functions disrupted to determine where precisely the regulation of metabolite flux resides.
Thermophilic bacilli capable of growth on methanol as the sole carbon substrate were isolated in pure culture and characterized metabolically. All strains possessed an nicotinamide adenine dinucleotide (NAD) linked methanol (alcohol) dehydrogenase (MDH) and generally grew on methanol with a very high efficiency. MDH had a low affinity for methanol but, along with reduced nicotinamide adenine dinucleotide (NADH) oxidase and hexulose phosphate synthase, was synthesized at very high levels. Similarly, Bacillus stearothermophilus synthesized glycerol kinase at extremely high levels and, when growing in glycerol-limited chemostat culture, had up to 40% (weight for weight) of its total protein as this 1 enzyme. Clearly, these organisms possess highly efficient expression systems. Studies of glucose metabolism in yeast revealed novel information on the role of cyclic adenosine monophosphate (cAMP) and protein phosphorylation in the regulation of glucose uptake and fermentation.
THE METHODOLOGIES APPLIED TO WORK ON THERMOTOLERANT METHYLOTROPHIC BACILLI AND THERMOPHILIC HETEROTROPHIC BACILLUS STEAROTHERMOPHILUS WILL BE CLOSELY SIMILAR. THESE WILL INVOLVE INITIAL QUANTITATIVE DETERMINATION OF FLUX RATES (OF METHANOL AND GLUCOSE, RESPECTIVELY) AND ANALYSIS OF FERMENTATION PRODUCTS WHEN ORGANISMS ARE GROWING IN RIGIDLY CONTROLLED ENVIRONMENTS IN CHEMOSTAT CULTURE. THIS WILL BE FOLLOWED BY THE APPLICATION OF GENETIC TECHNIQUES TO MODIFY SPECIFIC ENZYME ACTIVITIES AND THEREBY ENHANCE METABOLIC RATES.
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.
- natural sciences biological sciences biochemistry biomolecules lipids
- social sciences sociology governance environmental governance
- natural sciences chemical sciences organic chemistry alcohols
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
- engineering and technology industrial biotechnology bioprocessing technologies fermentation
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Coordinator
S10 2TN SHEFFIELD
United Kingdom
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