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New generation of biocatalysts with fractal supramolecular structure

Objectif

The main goal of the project is to bring into existence new generations of enzyme preparations with non-conventional supramolecular level of organization and to test their applicability at industrial scale.

Within this framework we are to pursue the following objectives:

- to elaborate scale-up-friendly techniques based on the use of 'smart' polymers for enzyme purification;
- to synthesize enzyme-based biocatalysts with diverse topological properties:
cross-linked enzyme aggregates (CLEAs) with statistical fractal structure;
- to investigate physicochemical properties of new enzyme preparations;
- to study chemical performance of new generation of biocatalysts;
- to test the applicability of novel enzyme preparations in large-scale processes.

Protein purification at a large scale
One of the basic directions of our experimental research would be the use of smart polymers loaded with ligands specific for enzymes of interest (penicillin acylases, lipases, thermolysin, oxynitrilases) for their purification. We expect this technique to be as specific as affinity chromatography and easy-to-scale-up in contrast to the latter. To treat this problem as thorough as possible, several teams with good experience in smart polymer synthesis and characterization, enzymology and bioprocess design are integrated.
Cross-Linked Enzyme Aggregates (CLEAs): extending supramolecular scopes of biocatalyst architecture
For application in biotechnology enzyme should be formulated, e.g. immobilized. Here we introduce a novel type of biocatalyst, CLEA, which possess a number of attractive properties:
Biocatalyst consists of nearly pure enzyme (and reminds in this respect cross-linked enzyme crystals, CLECs);
It is easy to synthesize (create precipitation conditions and perform cross-linking, in contrast to CLECs);
Its microstructure, being intrinsically fractal, is highly ordered in contrast to conventional enzyme preparations;
Catalytic performance at unnatural operational conditions (e.g. organic solvents);
Within the project we plan to elaborate new techniques for preparation of CLEA's with fractal supramolecular structure applicable to enzymes of interest and establish correlations between this structure and biocatalytic performance.
Applications of CLEAs in biotechnology.

The final destination of the project would be the attest of created biocatalysts for biotechnological purposes. With this respect in case of penicillin acylases we would concentrate on (-lactam antibiotic synthesis, for lipases - on resolution of chiral amines and alcohols, on peptide synthesis in organic media with thermolysin, and asymmetric synthesis of cyanohydrins with oxynitrilases. These studies would be conducted by teams qualified in enzymology, biotechnology, bioorganic chemistry and biophysics.

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Coordinateur

Delft University of Technology
Contribution de l’UE
Aucune donnée
Adresse
Julianalaan 136
2628 BL Delft
Pays-Bas

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Dernière mise à jour: 30 Octobre 2002

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Permalink: https://cordis.europa.eu/project/id/INTAS-2001-00673/fr

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