Synthesising Enzymes

Cloning and overexpression of these enzymes and alteration of their substrate specificity are under way as part of our goals to develop new chemical-enzymatic strategies for practical synthesis of sugar-related substances. We have developed several chemoenzymatic strategies for the synthesis of glycoproteins in order to study the role carbohydrates play in glycoprotein structure and function. A DERA-catalyzed sequential asymmetric aldol addition reaction is employed to make the epothilone precursors.Site-directed mutagenesis of proteases, intein-mediated transpeptidation and sugar-assisted ligation for condensation of glycopeptide segments and sugar -assisted glycopeptide ligation have been developed and used in the synthesis of homogeneous glycoproteins. Glycosyltransferases, engineered subtilisins, and self-splicing inteins are the key mediators in the synthesis. DERA accepts a broad range of acceptor and donor aldehydes in addition to the natural substrates.Since these 1,3-polyol systems are useful synthons, we have further examined the scope of this enzymatic methodology.

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Sugar or carbohydrate synthesis is important for the development of diagnostic tests, vaccines and new drugs.

In a study, researchers have synthesized carbohydrates with enzymes through a reaction that was not much studied so far on these biomolecules and which creates few by-products.

This is a variation from a reaction called front-face or SNi-like" says Carme Rovira.

"The advantage of this reaction in glycosidase -continues the researcher- is that it makes the synthesis in a clean way, without almost causing hydrolysis residues and using economically viable substrates (sugar)." "Basically, we used our experience with enzymes that synthesize carbohydrates to apply the reaction to one side in other enzymes with more biotechnological possibilities" concluded the researcher.

Our synthetic strategy emphasizes a combination of chemical and enzymatic methods, with particular focus on the use of enzymes for stereocontrolled processes.

Major efforts in synthetic carbohydrate chemistry include asymmetric aldol condensations for the synthesis of novel monosaccharides based on aldolases, and stereocontrolled synthesis of oligosaccharides and glycopeptides based on glycosyl transferases.

Sugar or carbohydrate synthesis in the laboratory (conventional chemical synthesis) is usually expensive and requires a lot of work; therefore lots of efforts have been made over the last years to do so with enzymes, which are natural catalyzers.

However, the enzymes that synthesize sugar (called glycosyltransferases) are hard to manipulate.


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