High yielding one-pot enzyme-catalyzed synthesis of UDP-glucose in gram scales

Uridine diphosphoglucose is an important cofactor of glucosylating enzymes. A simple and high yielding one-pot enzymatic synthesis of UDPG on a gram s cale from glucose via hexokinase, phosphoglucomutase and UDPG pyrophosphory lase (UGPase) is described. Repetitive addition of substrate was used to av oid inhibition of UGPase. The approach allows recovery of active enzymes an d their re-use. The synthesis of UDP-[4-C-13]-glucose on a 0.5 g scale resu lted in a final yield of 70% and a purity of > 95% after chromatographic pu rification. (C) 2001 Elsevier Science Ltd. All rights reserved.