Immobilization, stabilization and biocatalysis of selected enzymatic systems in non-conventional media

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The main objective of the applicant's research program is to advance and carry out experimental work that should contribute to the understanding of the mechanistic action and biocatalysis in non-conventional media (NCM), in particular organic solvent media (OSM) and solvent free media (SFM), of selected immobilized enzymes of microbial origin. Although the research work carried out by the applicant addressedthe immobilization and biocatalysis of lipoxygenase/hydroperoxide lyase, the stability of these enzymes as well as the elucidation of their mechanistic action in NCM are still among the major challenges to be overcome. The acquired knowledge and experience in our laboratory in immobilization and biocatalysis of enzymes in non-conventional media led us to move to novel horizons in research, in particular the use of novel nanotechnology approach for enzyme immobilization as well as the modeling and bioinformatics for setting the experimental design for the elucidation of their structure-function in non-conventional media. The specific objectives are: (a) to develop novel non-conventional media (NCM), including organic solvent media (OSM) and free-solvent media (FSM) for higher specificity, catalytic efficiency and stability, using substrate model compounds; (b) to investigate enzyme secondary structure-function relationship in the selected NCM by extensive conformational studies using spectroscopic techniques, including FTIR, circular dichroism and fluorescence for better understanding of their mechanistic actions; (c) to develop novel methodologies, in particular nanoencapsulation, for the immobilization and stabilization of the enzymatic activities of the selected microbial enzymatic extract; and (d) to use the response-surface methodology (RSM) as an approach for investigating the relationship between the catalytic properties of the immobilized enzymes and the nature of supports as well as their effect on internal and external diffusion and on the interfacial surface area or mass transfer.