engleski

Enzymatic cascade reaction for the synthesis of imino sugar precursors

Multistep enzyme-catalyzed reactions are gaining significant attention due to their low environmental impact and sustainability. One interesting approach to these reactions is the use of several different cell-free biocatalysts in one-pot (Systems Biocatalysis) which imitates the biosynthetic pathways in living cells. Such cascade reaction is investigated in this work. The first reaction step is the oxidation of amino alcohol catalyzed by alcohol dehydrogenase (ADH) coupled with coenzyme regeneration catalyzed by NADH oxidase (NOX). The second reaction step is the aldol addition of dihydroxyacetone to the oxidation product catalyzed by D-fructose-6-phosphate aldolase (FSA), to synthesize an imino aldol adduct. Our previous experience with similar system showed that such cascade reaction requires careful selection of the reaction conditions to achieve favorable substrate conversion and product yield. Thus, a detailed analysis of the reaction system was done. It included measuring the influence of substrate concentration, coenzyme concentration and ADH/NOX activity ratio on the reaction outcome. Also, enzyme operational stability during the reaction was investigated. At this point the highest diol conversion that was achieved was around 60 % at diol concentration of 20 mM.

enzymatic cascade reactions, imino sugars, aldol addition

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