RESULTS
WP1 - RATIONAL BIOPROSPECTING & CULTIVATONS
In the frame of the EU project “MICROSMETICS” a Rational Drug Design approach has been developed. Starting from the CosIng Repository of teh European Commision, an accurate functional prediction model was created for all known cosmetic functions. Additionally it was constructed the homology models of specific cosmetic target receptors (tyrosinase, elastase, hyluronidase and collagenase) for which the appropriate in vitro tests have already been developed. More than 40.000 known microbial metabolites were processed through a consensus scoring prediction protocol using: a) functional prediction model b) virtual screening procedure for the above 4 selected receptors c) similarity search based on all known molecules from literature that bind to the specific receptors and d) toxicological profile filtering. From the top hits metabolites, 100 microorgnanisms from global biodiversity that can produce those metabolites or analogues were selected to be cultivated. Among them 55 fungi and 55 actinomycetes were cultivated under “nutritional arrays”. Approximately 1100 extracts have been generated.
WP3 - DOWNSTREAMING
Results in progress
WP2 - BIOEVALUATION
A broad spectrum of bioassays and novel analytical approaches are being incorporated for the evaluation of anti-ageing, more specifically anti-oxidant, skin-protecting, and skin-whitening activity of all derived products. For the evaluation of antioxidant activity the DPPH and ABTS assays were used. Skin-protecting was evaluated by measuring spectrophotometrically the inhibitory properties of samples against enzymes which are related to the elasticity and moisture of the skin (elastase collagenase, and hyaluronidase). The skin whitening activities were determined by the tyrosinase assay, using L-DOPA as substrate. Finally for he safety of those extracts cytotoxicity was evaluated on A2058, CCD25sk, HepG2 cell lines by the ΜΤΤ method. The results of the above bioassays permitted the elimination of all toxic and non active extracts. Among the 1100 extracts, 100 were selected as highly potent and have been forwarded for LC-HRMS profiling and dereplication.
WP3 - DOWNSTREAMING
The top 100 extracts were selected and a strategy combining UHPLC/Orbitrap-HRMS, in positive and negative modes, with multivariate statistical methods was applied. All derived chromatograms have been analyzed and a positive correlation between the profiles of the extracts with the aforementioned bioassays was observed. Thus, the 10 most promising extracts that represent the clusters generated in metabolomics have been forwarded for large-scale cultivation and bioassay guided isolation of novel molecules.
WP4 - DEVELOPMENT
Results in progress