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Updated: May 25, 2026

Using Synthetic Biology to Engineer Living Cells That Interface with Programmable Materials
Published on: March 9, 2017
In-microbe formation of nucleotide sugars in engineered Escherichia coli
Ting Yang1, Yael Bar-Peled, James Amor Smith
1Complex Carbohydrate Research Center (CCRC), University of Georgia, Athens, GA 30602, USA.
Abstract:
Numerous different nucleotide sugars are used as sugar donors for the biosynthesis of glycans by bacteria, humans, fungi, and plants. However, many of these nucleotide sugars are not available either in their native form or with the sugar portion labeled with a stable or radioactive isotope. Here we demonstrate the use of Escherichia coli metabolically engineered to contain genes that encode proteins that convert monosaccharides into their respective monosaccharide-1-phosphates and subsequently into the corresponding nucleotide sugars. In this system, which we designated "in-microbe", reactions occur within 2 to 4 h and can be used to generate nucleotide sugars in amounts ranging from 5 to 12.5 μg/ml cell culture. We show that the E. coli can be engineered to produce the seldom observed nucleotide sugars UDP-2-acetamido-2-deoxy-glucuronic acid (UDP-GlcNAcA) and UDP-2-acetamido-2-deoxy-xylose (UDP-XylNAc). Using similar strategies, we also engineered E. coli to synthesize UDP-galacturonic acid (UDP-GalA) and UDP-galactose (UDP-Gal). ¹³C- and ¹⁵N-labeled NDP-sugars are formed using [¹³C] glucose as the carbon source and with [¹⁵N]NH₄Cl as the nitrogen source.
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