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One‐pot enzymatic production of dTDP‐4‐Keto‐6‐Deoxy‐ d ‐glucose from dTMP and glucose‐1‐phosphate

One‐pot enzymatic production of dTDP‐4‐Keto‐6‐Deoxy‐ d ‐glucose from dTMP and glucose‐1‐phosphate An enzymatic production method for dTDP‐4‐keto‐6‐deoxy‐d‐glucose, a key intermediate of various deoxysugars in antibiotics, was developed starting from dTMP, acetyl phosphate, and glucose‐1‐phosphate. Four enzymes, i.e., TMP kinase, acetate kinase, dTDP‐glucose synthase, and dTDP‐d‐glucose 4,6‐dehydratase' were overexpressed using T7 promoter system in the E. coli BL21 strain, and the dTDP‐4‐keto‐6‐deoxy‐d‐glucose was synthesized by using the enzyme extracts in one‐pot batch system. When 20 mM dTMP of initial concentration was used, Mg2+ ion, acetyl phosphate, and glucose‐1‐phosphate concentrations were optimized. About 95% conversion yield of dTDP‐4‐keto‐6‐deoxy‐d‐glucose was obtained based on initial dTMP concentration at 20 mM dTMP, 1 mM ATP, 60 mM acetyl phosphate, 80 mM glucose‐1‐phosphate, and 20 mM MgCl2. The rate‐limiting step in this multiple enzyme reaction system was the dTDP‐glucose synthase reaction. Using the reaction scheme, about 1 gram of purified dTDP‐4‐keto‐6‐deoxy‐d‐glucose was obtained in an overall yield of 81% after two‐step purification, i.e., anion exchange chromatography and gel filtration. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 84: 452–458, 2003. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biotechnology and Bioengineering Wiley

One‐pot enzymatic production of dTDP‐4‐Keto‐6‐Deoxy‐ d ‐glucose from dTMP and glucose‐1‐phosphate

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References (11)

Publisher
Wiley
Copyright
Copyright © 2003 Wiley Periodicals, Inc., A Wiley Company
ISSN
0006-3592
eISSN
1097-0290
DOI
10.1002/bit.10789
pmid
14574703
Publisher site
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Abstract

An enzymatic production method for dTDP‐4‐keto‐6‐deoxy‐d‐glucose, a key intermediate of various deoxysugars in antibiotics, was developed starting from dTMP, acetyl phosphate, and glucose‐1‐phosphate. Four enzymes, i.e., TMP kinase, acetate kinase, dTDP‐glucose synthase, and dTDP‐d‐glucose 4,6‐dehydratase' were overexpressed using T7 promoter system in the E. coli BL21 strain, and the dTDP‐4‐keto‐6‐deoxy‐d‐glucose was synthesized by using the enzyme extracts in one‐pot batch system. When 20 mM dTMP of initial concentration was used, Mg2+ ion, acetyl phosphate, and glucose‐1‐phosphate concentrations were optimized. About 95% conversion yield of dTDP‐4‐keto‐6‐deoxy‐d‐glucose was obtained based on initial dTMP concentration at 20 mM dTMP, 1 mM ATP, 60 mM acetyl phosphate, 80 mM glucose‐1‐phosphate, and 20 mM MgCl2. The rate‐limiting step in this multiple enzyme reaction system was the dTDP‐glucose synthase reaction. Using the reaction scheme, about 1 gram of purified dTDP‐4‐keto‐6‐deoxy‐d‐glucose was obtained in an overall yield of 81% after two‐step purification, i.e., anion exchange chromatography and gel filtration. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 84: 452–458, 2003.

Journal

Biotechnology and Bioengineering – Wiley

Published: Nov 20, 2003

Keywords: dTDP‐4‐keto‐6‐deoxy‐ d ‐glucose; TMP kinase; acetate kinase; dTDP‐glucose synthase; dTDP‐ d ‐glucose 4,6‐dehydratase

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