Journal
CARBOHYDRATE RESEARCH
Volume 345, Issue 11, Pages 1622-1626Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carres.2010.04.025
Keywords
UDP-Glc; Biocatalytic synthesis; One-pot; Magnetic nanoparticles; Immobilized biocatalyst
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Uridine diphosphate glucose (UDP-Glc) serves as a glucosyl donor in many enzymatic glycosylation processes. This paper describes a multiple enzyme, one-pot, biocatalytic system for the synthesis of UDP-Glc from low cost raw materials: maltodextrin and uridine triphosphate. Three enzymes needed for the synthesis of UDP-Glc (maltodextrin phosphorylase, glucose-1-phosphate thymidylytransferase, and pyrophosphatase) were expressed in Escherichia coli and then immobilized individually on amino-functionalized magnetic nanoparticles. The conditions for biocatalysis were optimized and the immobilized multiple-enzyme biocatalyst could be easily recovered and reused up to five times in repeated syntheses of UDP-Glc. After a simple purification, approximately 630 mg of crystallized UDP-Glc was obtained from 1l of reaction mixture, for a moderate yield of around 50% (UTP conversion) at very low cost. (C) 2010 Elsevier Ltd. All rights reserved.
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