New Carbohydrate Oxidising Enzymes prepared by directed evolution and their application in synthesis of AminoGlycoside derivatives with antibiotic potential and rare sugars.

Carbohydrate-oxidising enzymes (COE) are a group of enzymes which are active on specific carbohydrates and can use molecular oxygen as a mild oxidant. Oxidation is typically directed towards specific positions of the sugar ring.

Regioselectivity and substrate specificity relies on the architecture of active site, therefore the direct evolution approach in this project will allow to generate a toolbox of effective and regiospecific catalysts to modify various carbohydrate scaffolds.

Our aim is to use this toolbox to generate a library of bioactive compounds: i) aminoglycoside derivatives starting with analogues of Kanamycin as candidates to help in the fight against the global problem of antimicrobial resistance.

This process in combination with a further chemical step may lead to a large library of derivatives prepared in short time with only a two-step chemoenzymatic process; ii) rare monosaccharides and oligosaccharides as promising candidates with health benefits. Antimicrobial activities of prepared library of compounds will be measured and statistically evaluated.

Furthermore, this project will advance the current knowledge of COE which is of utmost interest for fundamental biology and crucial for industrial biotechnology and chemistry applications.

Biocatalysis will be applicable to the synthesis of a wide range of saccharide related compounds, solving one of the big challenges in carbohydrate chemistry, the regioselective differentiation of the OH groups, in a green, sustainable and very efficient way.