In the frame of TASCMAR, an H2020 EU-funded project, more than 280 invertebrates and 400 microorganisms were collected from the under-investigated mesophotic zone (between 30 and 100 meters’ depth) of the Indian ocean, the Red sea and the Mediterranean.
The invertebrates were lyophilized and extracted by Accelerated Solvent Extraction (ASE) methodology using a mixture of dichloromethane and methanol (50:50, v/v). The microorganisms were cultivated under solid and liquid conditions in Marine Broth in 1L scale, using in-situ solid phase extraction technology and they were extracted with AcOEt and MeOH . All 1370 extracts were evaluated for their anti-aging and skin-whitening activity using elastase and tyrosinase enzymatic assays.
According to the results for the invertebrates; 12.4% presented activity against elastase and 16.3% against tyrosinase and 0.5% of the microorganisms showed tyrosinase and elastase inhibitory activity.
Six invertebrates (Iotrochota sp.1, Iotrochota sp.2, Biemna sp., Terpios sp., Pseudoceratina sp., Thrinacophora sp.) and one microorganism ( Cladosporium halotolerans) were selected for further investigation. HRMS techniques were employed in order to investigate the metabolites present in each extract. Thus various analytical chemistry techniques and software, such as the R package xMSannotator using a custom library from the Dictionary of Natural Product and a combining strategy of Molecular Networking and In silico MS/MS fragmentation pipeline using the Universal Natural Products Database, were employed for dereplication .
The extracts were fractionated by Medium Pressure Liquid Chromatography (MPLC), Flash Chromatography or Centrifugal Partition Chromatography (CPC) using various solvent systems. All the fractions were further investigated for their biological activity. Bio-guided selection of the most active fractions was followed by metabolic profiling, structure elucidation and isolation of the high added value marine compounds by NMR and LC-MS techniques.
Acknowledgement: TASCMAR project ( www.tascmar.eu ) has been funded by the European Union in the frame of H2020 (Grant Agreement No 634674).
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