Objective
THE LONG TERM OBJECTIVE OF THE JOINT PROJECT IS TO WORK OUT ECONOMIC METHODS FOR THE CONTINUOUS ENZYMATIC SYNTHESIS OF HIGH-VALUE FINE CHEMICALS BY USING REDOX REACTION REQUIRING NAD(H), FAD(H) OR ARTIFICIAL DYES AS COFACTOR.
THE DEVELOPMENT OF THE FOUR DIFFERENT SYSTEMS STUDIED REQUIRES THE DESIGN OF NEW MEMBRANE-REACTORS BOTH IS AQUEOUS OR ORGANIC ENVIRONMENT.
THE PRODUCTION OF THE VALUABLE CHEMICALS IS EXPECTED :
1. 12-KETOCHENODEOXYCHOLIC AND 12 KETOURSODEOXY CHOLIC ACID WHICH ARE EXPENSIVE PRECURSORS FOR THE CHEMICAL SYNTHESIS OF DRUGS TO TREAT GALLSTONE ALIMET,
2. GLUCONIC ACID AND MANNITOL FOR AGROFOOD INDUSTRY,
3. L-ASCORBIC ACID (VITAMIN C) FOR PHARMACEUTICAL INDUSTRY,
4. NEUTRAL STEROIDS AND LONG CHAIN ALCOHOLS FOR PHARMACEUTICAL AND AGROFOOD INDUSTRY.
Microbial nicotinamide adenine dinucleotide (phosphate) (reduced) (NAD(P)(H)) dependent (dep) dehydrogenases (DH) are of great potential for the synthesis of fine chemicals (chiral synthons, steroids, carbohydrates, optically pure compounds (labelled) or pharmaceutical precursors). To exploit their catalytic potential 2 main criteria must be fulfilled: optimized bioprocessing methods and downstream processing methods (production, purification, adaptation to large scale); appropriate reactor technology for economic use of enzymes and coenzymes considering enzyme stability and coenzyme regeneration.
Research into the methodology has given thefollowing results:
development of a simplified synthesis of N6-(2-aminoethyl)-nicotinamide adenine dinucleotide, N6-(2-aminoethyl)-nicotinamide adenine dinucleotide phosphate and, as spin off, N6-(2-aminoethyl)-flavin adenine dinucleotide (mild conditions, yield 15 to 40%;
nicotinamide adenine dinucleotide phosphate (reduced)-dep 12 alpha-hydroxy steroid-DH (Clostridium group P) gave a cheap process for production (0.9 E6 U/900 l) and total purification (130 U/mg, yield 53%), enzyme characterization and laboratory scale synthesis of 12-ketochenodeoxycholic acid;
nicotinamide adenine dinucleotide (reduced)-dep L-phenylatanine-DH (Sporosarcina urae) gave a bioprocess optimization (2E5 U/50 l), downstream processing by 2-step affinity chromatography (100 U/mg, yield 70%) and full characterization and synthesis of 2-15 NH2-phenylalanine and 2-15 NH2-tyrosine are in progress;
with nicotinamide adenine dinucleotide phosphate (reduced)-dep glucose-DH (Cryptococcus uniguttulatus) native nicotinamide adenine dinucleotide phosphate and N6-functionalized nicotinamide adenine dinucleotide phosphate (low molecular and high molecular) are equally accepted by glucose-DH;
nicotinamide adenine dinucleotide phosphate (reduced)-dep sec alcohol-DH (Thermoanaerobium brockii) on a laboratory scale gives the synthesis of S-(+)-sulcatol, (4R-2H)-reduced nicotinamide adenine dinucleotide phosphate and R-3-bromo-5-(l-hydroxy-2-bromoethyl) isoxazole (broxaterol precursor);
nicotinamide adenine dinucleotide (reduced)-dep mannitol-DH (Saccharomyces cerevisiae) gave medium optimization (2.95E4 U/220 l) purification (36 U/mg) and characterization nicotinamide adenine dinucleotide (phosphate) (reduced) scale simultaneous mannitol and gluconic acid production;
retention of nicotinamide adenine dinucleotide (phosphate) (reduced) by a negatively charged urea formaldehyde (UF) membrane (M cut off 2000) with retention greater than 99% and 99.9% under process conditions.
DEVELOPMENT OF CONJUGATED ENZYMATIC PROCESSES YIELDING TWO REQUIRED PRODUCTS WITH REGENERATION OF COFACTOR.
IN PARTICULAR :
1. THE SIMULTANEOUS PRODUCTION OF GLUCONIC ACID AND MANNITOL WILL BE IMPROVED BY DEVELOPMENT OF SUITABLE NAD DERIVATIVES OR REDOX DYES SUBSTITUTES.
2. A NEW PROCESS WILL BE DEVELOPPED FOR PRODUCTION OF GLUCONIC ACID FROM GLUCOSE BY GLUCOSE OXIDASE FREED OF ITS BOUND-FAD AND REPLACED BY FAD ANALOGUES, REGENERATED BY CATALASE.
3. MULTIENZYMATIC PROCESSES FOR THE SYNTHESIS OF L-ASCORBIC ACID (VITAMIN C) BY INTRASEQUENTIAL COFACTOR REGENERATION.
4. SCREENING, PURIFICATION, CHARACTERIZATION AND STABILIZATION OF 9 DIFFERENT OXYDOREDUCTASES.
5. DEVELOPMENT OF NEW MEMBRANES AND MEMBRANE REACTORS FOR NON-CONVENTIONAL ENVIRONMENT AND COFACTOR REGENERATION.
6. DEVELOPMENT OF DOWN STREAM SEPARATION PROCEDURES BASED ON MEMBRANETECHNOLOGY AND ABSORPTIVE OR ION EXCHANGE PROCESSES.
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 chemical sciences organic chemistry aldehydes
- natural sciences chemical sciences organic chemistry organic reactions
- natural sciences biological sciences biochemistry biomolecules carbohydrates
- natural sciences chemical sciences organic chemistry alcohols
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
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Coordinator
80636 MUENCHEN
Germany
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