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Determinants of the nucleotide specificity in the carbohydrate epimerase family 1

Stevie Van Overtveldt, Matthieu Da Costa, Ophelia Gevaert, Henk-Jan Joosten, Koen Beerens (UGent) and Tom Desmet (UGent)
(2020) BIOTECHNOLOGY JOURNAL. 15(11).
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Abstract
In recent years, carbohydrate epimerases have attracted increasing attention as promising biocatalysts for the production of specialty sugars and derivatives. The vast majority of these enzymes are active on nucleotide-activated sugars, rather than on their free counterparts. Although such epimerases are known to have a clear preference for a particular nucleotide (UDP, GDP, CDP, or ADP), very little is known about the determinants of the respective specificities. In this work, sequence motifs are identified that correlate with the different nucleotide specificities in one of the main epimerase superfamilies, carbohydrate epimerase 1 (CEP1). To confirm their relevance, GDP- and CDP-specific residues are introduced into the UDP-glucose 4-epimerase fromThermus thermophilus, resulting in a 3-fold and 13-fold reduction inK(M)for GDP-Glc and CDP-Glc, respectively. Moreover, several variants are severely crippled in UDP-Glc activity, which further underlines the crucial role of the identified positions. Hence, the analysis should prove to be valuable for the further exploration and application of epimerases involved in carbohydrate synthesis.
Keywords
Molecular Medicine, Applied Microbiology and Biotechnology, General Medicine, carbohydrate epimerase, CDP-paratose 2-epimerase, nucleotide sugar, rational mutagenesis, UDP-glucose 4-epimerase, SUCROSE SYNTHASE, VITAMIN-C, BIOSYNTHESIS, ENZYME, GLYCOSYLTRANSFERASE, TYVELOSE, IDENTIFICATION, PURIFICATION, MECHANISM, EVOLUTION

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MLA
Van Overtveldt, Stevie, et al. “Determinants of the Nucleotide Specificity in the Carbohydrate Epimerase Family 1.” BIOTECHNOLOGY JOURNAL, vol. 15, no. 11, 2020, doi:10.1002/biot.202000132.
APA
Van Overtveldt, S., Da Costa, M., Gevaert, O., Joosten, H.-J., Beerens, K., & Desmet, T. (2020). Determinants of the nucleotide specificity in the carbohydrate epimerase family 1. BIOTECHNOLOGY JOURNAL, 15(11). https://doi.org/10.1002/biot.202000132
Chicago author-date
Van Overtveldt, Stevie, Matthieu Da Costa, Ophelia Gevaert, Henk-Jan Joosten, Koen Beerens, and Tom Desmet. 2020. “Determinants of the Nucleotide Specificity in the Carbohydrate Epimerase Family 1.” BIOTECHNOLOGY JOURNAL 15 (11). https://doi.org/10.1002/biot.202000132.
Chicago author-date (all authors)
Van Overtveldt, Stevie, Matthieu Da Costa, Ophelia Gevaert, Henk-Jan Joosten, Koen Beerens, and Tom Desmet. 2020. “Determinants of the Nucleotide Specificity in the Carbohydrate Epimerase Family 1.” BIOTECHNOLOGY JOURNAL 15 (11). doi:10.1002/biot.202000132.
Vancouver
1.
Van Overtveldt S, Da Costa M, Gevaert O, Joosten H-J, Beerens K, Desmet T. Determinants of the nucleotide specificity in the carbohydrate epimerase family 1. BIOTECHNOLOGY JOURNAL. 2020;15(11).
IEEE
[1]
S. Van Overtveldt, M. Da Costa, O. Gevaert, H.-J. Joosten, K. Beerens, and T. Desmet, “Determinants of the nucleotide specificity in the carbohydrate epimerase family 1,” BIOTECHNOLOGY JOURNAL, vol. 15, no. 11, 2020.
@article{8682797,
  abstract     = {{In recent years, carbohydrate epimerases have attracted increasing attention as promising biocatalysts for the production of specialty sugars and derivatives. The vast majority of these enzymes are active on nucleotide-activated sugars, rather than on their free counterparts. Although such epimerases are known to have a clear preference for a particular nucleotide (UDP, GDP, CDP, or ADP), very little is known about the determinants of the respective specificities. In this work, sequence motifs are identified that correlate with the different nucleotide specificities in one of the main epimerase superfamilies, carbohydrate epimerase 1 (CEP1). To confirm their relevance, GDP- and CDP-specific residues are introduced into the UDP-glucose 4-epimerase fromThermus thermophilus, resulting in a 3-fold and 13-fold reduction inK(M)for GDP-Glc and CDP-Glc, respectively. Moreover, several variants are severely crippled in UDP-Glc activity, which further underlines the crucial role of the identified positions. Hence, the analysis should prove to be valuable for the further exploration and application of epimerases involved in carbohydrate synthesis.}},
  articleno    = {{2000132}},
  author       = {{Van Overtveldt, Stevie and Da Costa, Matthieu and Gevaert, Ophelia and Joosten, Henk-Jan and Beerens, Koen and Desmet, Tom}},
  issn         = {{1860-6768}},
  journal      = {{BIOTECHNOLOGY JOURNAL}},
  keywords     = {{Molecular Medicine,Applied Microbiology and Biotechnology,General Medicine,carbohydrate epimerase,CDP-paratose 2-epimerase,nucleotide sugar,rational mutagenesis,UDP-glucose 4-epimerase,SUCROSE SYNTHASE,VITAMIN-C,BIOSYNTHESIS,ENZYME,GLYCOSYLTRANSFERASE,TYVELOSE,IDENTIFICATION,PURIFICATION,MECHANISM,EVOLUTION}},
  language     = {{eng}},
  number       = {{11}},
  pages        = {{8}},
  title        = {{Determinants of the nucleotide specificity in the carbohydrate epimerase family 1}},
  url          = {{http://doi.org/10.1002/biot.202000132}},
  volume       = {{15}},
  year         = {{2020}},
}
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