Research objectives and content
Mechanisms affecting the operational stability of enzymes in low-water environment will be analysed, with specific reference to enzymatic systems of potential, industrial interest. New experimental procedures will be developed to study the influence of each mechanisms. In particular, dehydration phenomena will be studied on immobilised samples of enzyme in a continuous, low-water reactor. A new experimental procedure will be followed, based on a modified 0-18 labelling technique. Moreover, covalent changes in the enzyme molecules will be detected by electrospray mass spectrometry. The influence of intermolecular reactions between proteins, as well as the effect of covalent oxidative processes, will be studied through specific tests. An original approach will be developed to analyse conformational changes on low-water enzymes: proteins will be immobilised on the outside surface of an optical fibre. Fluorescence and adsorption properties could be studied from the inside.
Packed beds will be used as experimental reactors. They will be run in differential mode, in order to expose the biocatalyst to constant operating conditions. Different reactor configurations (single reactor, reactors in series or in parallel) will be tested in order to monitor accurately catalyst activity, as well as to prevent experimental problems. Experimental approaches will be developed to obtain biocatalytic nonaqueous systems offering good performance in terms of catalytic activity, long-term stability and product yield. To this respect, several immobilisation methods and molecular imprinting techniques will be tested. Training content (objective, benefit and expected impact)
The researcher will acquire a wider competence in a scientific area of high industrial interest. After the conclusion of the stage, research activities will be continued within the Universita Federico II in Naples (university of origin, in a less-favoured region). Further cooperation between Universita Federico II and University of Strathclyde will be encouraged.
Links with industry / industrial relevance (22)
Enzyme catalysis in low-water systems has resulted in a large number of potential applications, e.g. peptide synthesis, biosurfactant synthesis, interesterification of oils and fats, clinical analysis, polymer chemistry. A better knowledge of the catalytic properties of enzymes in an organic milieu could expand the industrial use of such systems. The host research group has formal collaborative links with research laboratories of Unilever (Colworth, UK, and Vlaardingen, NL), and close contacts with other european industrial research labs, particularly Zeneca (UK), Degussa (D) and Novo-Nordisk (DK).