engleski

Multistep enzymatic synthesis of nucleotide-activated sugars

Four-step synthesis of UDP-Glc was performed and synthesized UDP-Glc was used as substrate in production of UDP-GlcUA. In the synthesis of UDP-Glc, isotope-labeling did not have any influence on the reaction and similar yields were obtained with unlabeled and labeled UDP-Glc. Although UDP-Glc was produced in the 3rd reaction, four-step reaction was performed. Since pyrophosphate affects yield of produced UDP-Glc, 4threaction was performed. Removal of a pyrophosphate dragged equilibrium toward UDP-Glc synthesis. Synthesis of UDP-Glc was very successful with theoretical yield between 86-99%. Conversion of UDP-Glc into UDP-GlcUA by human UDP-glucose dehydrogenase was performed with yield of approximately 100%. Produced UDP-GlcUA was purified as ammonium salt and exchanged into sodium salt. Both salts were used for reaction catalyzed by UDP-apiose/xylose synthase (AXS). In AXS catalyzed reaction with ammonium salt less UDP-Api was formed than in the reaction with sodium salt. AXS accepted all substrates, unlabeled and labeled UDP-GlcUA.

synthesis of UDP-glucose; production of UDP-glucuronic acid; UDP-glucuronic acid purification; UDP-Api/Xyl synthase reaction

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