Document Type

Article

Publication Date

3-2010

Publication Title

Journal of Molecular Catalysis B: Enzymatic

Abstract

Production of deoxy-thymidine-diphosphate (TDP)-sugars as substrates of glycosyltransferases, has been one of main hurdles for combinatorial antibiotic biosynthesis, which combines sugar moiety with aglycon of various antibiotics. Here, we report the one-pot enzymatic synthesis of TDP-2-deoxy-glucose employing high efficient TMP kinase (TMK; E.C. 2.7.2.12), acetate kinase (ACK; E.C. 2.7.1.21), and TDP-glucose synthase (TGS; E.C. 2.7.7.24) with phosphomannomutase (PMM; E.C. 5.4.2.8). In this study, replacing phosphoglucomutase (PGM; E.C. 5.4.2) by PMM from Escherichia coli gave four times higher specific activity on 2-deoxy-6-phosphate glucose, suggesting that the activity on 2-deoxy-glucose-6-phosphate was mainly affected by PMM activity, not PGM activity. Using an in vitro system starting from TMP and 2-deoxy-glucose-6-phosphate glucose, TDP-2-deoxy-glucose (63% yield) was successfully synthesized. Considering low productivity of NDP-sugars from cheap starting materials, this paper showed how production of NDP-sugars could be enhanced by controlling mutase activity.

Keywords

TDP-2-deoxy-glucose, Phosphomannomutase, Enzymatic synthesis, TMP, One-pot reaction

Volume

62

Issue

3-4

First Page

282

Last Page

287

DOI

https://doi.org/10.1016/j.molcatb.2009.11.008

ISSN

1381-1177

Comments

Originally published in Journal of Molecular Catalysis B: Enzymatic, Volume 62, Issues 3–4, March 2010, Pages 282-287, Elsevier.

https://doi.org/10.1016/j.molcatb.2009.11.008

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