One-pot enzymatic synthesis of UDP-D-glucose from UMP and glucose-1-phosphate using an ATP regeneration system.
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.
Lee
Hei Chan
HC
Institute of Biomolecule Reconstruction, Sun Moon University, Chungnam 336-708, Korea.
Lee
Seung-Don
SD
Sohng
Jae Kyung
JK
Liou
Kwangkyoung
K
D000255:Adenosine Triphosphate / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005958:Glucosephosphates / Q000378:metabolism*; D015394:Molecular Structure; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000235:genetics / Q000378:metabolism*; D014532:Uridine Diphosphate Glucose / Q000737:chemistry / Q000378:metabolism*; D014542:Uridine Monophosphate / Q000378:metabolism*
D029968:Escherichia coli Proteins; D005958:Glucosephosphates; D000255:Adenosine Triphosphate; C031590:glucose-1-phosphate; D014542:Uridine Monophosphate; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase; D014532:Uridine Diphosphate Glucose