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One-pot Enzymatic Synthesis of UDP-D-glucose from UMP and Glucose-1-phosphate Using an ATP Regeneration System

  • Lee, Hei-Chan (Institute of Biomolecule Reconstruction, Sun Moon University) ;
  • Lee, Seung-Don (Institute of Biomolecule Reconstruction, Sun Moon University) ;
  • Sohng, Jae-Kyung (Institute of Biomolecule Reconstruction, Sun Moon University) ;
  • Liou, Kwang-Kyoung (Institute of Biomolecule Reconstruction, Sun Moon University)
  • Published : 2004.07.31

Abstract

Glucose-1-phosphate uridylyltransferase from E. coli K12 was used to convert uridine-5'-triphosphate and glucose-1-phosphate to UDP-D-glucose. The conversion was efficient and completed within 5 minutes under the employed conditions. In addition, thymidine-5'-monophosphate kinase and acetate kinase were proven to be non-specific, converting udridine-5'-monophosphate to uridine-5'-triphosphate with 55% conversion after 6 h, which was much slower than the production of TTP under the same conditions (complete conversion within one hour). Since these two reactions could proceed under the same conditions, a one-pot synthesis of UDP-D-glucose with ATP regeneration was designed from easily available starting materials, and conversion up to 40% by HPLC peak integration was achieved given a reaction time of 4 h.

Keywords

References

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