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Article

Misincorporation of Galactose by Chondroitin Synthase of Escherichia coli K4: From Traces to Synthesis of Chondbiuronan, a Novel Chondroitin-Like Polysaccharide

1
HTL Biotechnology, 35133 Javene, France
2
CNRS, CERMAV, University Grenoble Alpes, 38000 Grenoble, France
*
Author to whom correspondence should be addressed.
Biomolecules 2020, 10(12), 1667; https://doi.org/10.3390/biom10121667
Received: 25 November 2020 / Revised: 8 December 2020 / Accepted: 10 December 2020 / Published: 12 December 2020
Chondroitin synthase KfoC is a bifunctional enzyme which polymerizes the capsular chondroitin backbone of Escherichia coli K4, composed of repeated β3N-acetylgalactosamine (GalNAc)-β4-glucuronic acid (GlcA) units. Sugar donors UDP-GalNAc and UDP-GlcA are the natural precursors of bacterial chondroitin synthesis. We have expressed KfoC in a recombinant strain of Escherichia coli deprived of 4-epimerase activity, thus incapable of supplying UDP-GalNAc in the bacterial cytoplasm. The strain was also co-expressing mammal galactose β-glucuronyltransferase, providing glucuronyl-lactose from exogenously added lactose, serving as a primer of polymerization. We show by the mean of NMR analyses that in those conditions, KfoC incorporates galactose, forming a chondroitin-like polymer composed of the repeated β3-galactose (Gal)-β4-glucuronic acid units. We also show that when UDP-GlcNAc 4-epimerase KfoA, encoded by the K4-operon, was co-expressed and produced UDP-GalNAc, a small proportion of galactose was still incorporated into the growing chain of chondroitin. View Full-Text
Keywords: chondroitin; K4 chondroitin synthase; NDP-sugar misincorporation chondroitin; K4 chondroitin synthase; NDP-sugar misincorporation
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MDPI and ACS Style

Leroux, M.; Michaud, J.; Bayma, E.; Armand, S.; Drouillard, S.; Priem, B. Misincorporation of Galactose by Chondroitin Synthase of Escherichia coli K4: From Traces to Synthesis of Chondbiuronan, a Novel Chondroitin-Like Polysaccharide. Biomolecules 2020, 10, 1667. https://doi.org/10.3390/biom10121667

AMA Style

Leroux M, Michaud J, Bayma E, Armand S, Drouillard S, Priem B. Misincorporation of Galactose by Chondroitin Synthase of Escherichia coli K4: From Traces to Synthesis of Chondbiuronan, a Novel Chondroitin-Like Polysaccharide. Biomolecules. 2020; 10(12):1667. https://doi.org/10.3390/biom10121667

Chicago/Turabian Style

Leroux, Mélanie, Julie Michaud, Eric Bayma, Sylvie Armand, Sophie Drouillard, and Bernard Priem. 2020. "Misincorporation of Galactose by Chondroitin Synthase of Escherichia coli K4: From Traces to Synthesis of Chondbiuronan, a Novel Chondroitin-Like Polysaccharide" Biomolecules 10, no. 12: 1667. https://doi.org/10.3390/biom10121667

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