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CORDIS - Resultados de investigaciones de la UE
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Contenido archivado el 2024-04-15

BIOCONVERSION OF HYDROPHILIC AND HYDROPHOBIC COMPOUNDS BY ENZYME SYSTEMS

Objetivo

DEVELOPMENT OF NEW ENZYME REACTORS FOR CONTINUOUS PRODUCTIONS OF WATER SOLUBLE AND WATER INSOLUBLE PRODUCTS REQUIRING REGENERATION OF REDOX COFACTORS.

THE FOLLOWING CHEMICALS WILL BE PRODUCED :

1. L-ALANINE, L-METHIONINE,
2. LONG CHAIN ALDEHYDES,
3. LACTIC ACID FROM MALIC ACIC,
4. HIGH-ADDED-VALUE LIPIDS.
The development of many industrial enzymatic processes is dependent upon the discovery of thermostable enzymes (ie those extracted from extremophiles) or methodologies for activity stabilization (ie immobilization) in the presence of organic solvents, with coenzyme regeneration.
Research was carried out in order to contribute to solving the general problem of enzyme stability and coenzyme regeneration in aqueous or mixed solvent and water solutions, in enzyme reactors and in liquid membranes.

The potentiality of some extremophilic organisms' cells and enzymes which are thermostable and resistant to common protein denaturing agents and to organic solvents was studied. These enzymes were purified and produced by genetic engineering.
A new alcohol dehydrogenase (ADH), a beta-galactosidase and a malic enzyme were purified from the extremophile Sulfolobus solfataricus. These thermostable and solvent resistant enzymes were used for the synthesis of long chain aldehydes, chiral compounds, L-amino acids and for malic acid conversion and lactose hydrolysis.

2 gene banks were constructed and the beta-galactosidase gene was isolated from the extremophile, cloned, sequenced and expressed in Escherichia coli. New types of solid gas bioreactors were developed and have been used with coenzyme regeneration for the synthesis of aldehydes or transesterification reactions catalyzed by lipases, whose activities and regioselectivities were investigated in different systems (eg microemulsions, etc) with and without immobilization. L-amino acid and aldehyde productions were investigated in liquid membranes with coenzyme regeneration.
Extremophile enzymes appear to be different with little or no homology with the corresponding ones from mesophiles; they are thermostable and solvent resistant proteins whose structures can be used as models for producing proteins by protein engineering.
STUDY OF THE TRANSESTERIFICATION AND SYNTHETIC REACTIONS OF LIPASES FROM DIFFERENT SOURCES FOR THE PRODUCTION OF HIGH ADDED VALUE LIPIDS FROM OLIVE OIL.

IN PARTICULAR IT IS PLANNED :

1. TO MODIFY LIPASES CHEMICALLY TO OBTAIN "HYDROPHOBIC LIPASES";
2. TO STUDY THE STRUCTURE-FUNCTION RELATIONSHIP OF LIPASES IN CHEMICALLY MODIFIED OR THERMOSTABLE ENZYMES;
3. TO USE ESR TECHNIQUES TO STUDY THE CONFORMATIONAL CHANGES OF LIPASES AND THEIR ROLE ON CATALYTIC ACTIVITY IN MICROEMULSIONS;
4. TO IMMOBILIZE LIPASES IN SOLUBLE POLYMERS OR IN SOLID STATE (SINGLE PROTEIN CRYSTAL).

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Coordinador

NATIONAL HELLENIC RESEARCH FOUNDATION
Aportación de la UE
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Dirección
48 VAS. CONSTANTINOU AVE.
11635 ATHINAI
Grecia

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