Advanced Development of Sugar Nucleotide Regeneration Cascade

Abstract

Sugar nucleotides are essential building blocks for the glycobiology of all organisms being intrinsically linked to cellular metabolism. However, the market price of sugar nucleotides is currently very high due to the inexistence of cost-effective manufacturing processes. In this thesis, an in vitro enzyme process for the synthesis of sugar nucleotides UDP-Man, UDP- Glc and UDP-GlcNAc using low cost substrates was developed and studied in order to decrease the manufacturing costs of sugar nucleotides. The effects of pH and buffer composition on the cascades were investigated leading to an increase in the productivity of the target product. Another importante discovery was the enzymatic production of the sugar nucleotide, UDP-Man, representing the first step towards the more economical production of this sugar. Moreover, it was found that, by adjusting the concentration of the enzymes in the cascades, the productivity of the UDP-Glc and the UDP-GlcNAc cascade could be increased by 52% and 56%, respectively. Finally, it was observed that when high concentration of product if formed, in situ removal of sugar nucleotide might be necessary. Given the results obtained, this thesis presents new data and ideas towards the establishment of a cost-effective in vitro enzymatic production process of UDP-Man, UDP-Glc and UDP-GlcNAc with very promising practical applications.