Catalytic properties and crystal structure of UDP-galactose 4-epimerase-like l-threonine 3-dehydrogenase from Phytophthora infestans
•A UDP-galactose 4-epimerase-like l-threonine 3-dehydrogenase (GalE-like L-ThrDH) from Phytophthora infestans, a plant pathogenic fungus, was successfully isolated and characterized.•The N-acetylglycine, an inhibitor of P. infestans GalE-like L-ThrDH, is a potential lead compound to develop new high...
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Veröffentlicht in: | Enzyme and microbial technology 2020-10, Vol.140, p.109627-109627, Article 109627 |
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Sprache: | eng |
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Zusammenfassung: | •A UDP-galactose 4-epimerase-like l-threonine 3-dehydrogenase (GalE-like L-ThrDH) from Phytophthora infestans, a plant pathogenic fungus, was successfully isolated and characterized.•The N-acetylglycine, an inhibitor of P. infestans GalE-like L-ThrDH, is a potential lead compound to develop new highly specific agrochemicals against P. infestans.•The hexameric GalE-like L-ThrDH was determined using the molecular replacement method at a resolution of 2.3 Å, and the binding mode and inhibition mechanism of N-acetylglycine were elucidated.
We report, for the first time, the three-dimensional structure and biochemical properties of a UDP-galactose 4-epimerase-like l-threonine 3-dehydrogenase (GalE-like L-ThrDH) from Phytophthora infestans, a plant disease-causing fungus. We identified GalE-like L-ThrDH using Kyoto Encyclopedia of Genes and Genomes (KEGG) database as a candidate target for the development of a new fungicide. The GalE-like L-ThrDH gene was expressed in Escherichia coli, and its product was purified and characterized. N-Acetylglycine was found to act as a competitive inhibitor of the enzyme (Ki =0.18 mM). The crystal structure of the unique hexameric GalE-like L-ThrDH was determined using the molecular replacement method at a resolution of 2.3 Å, in the presence of NAD+ and citrate, an analogue of the substrate. Based on the molecular docking simulation, N-acetylglycine molecule was modeled into the active site and the binding mode and inhibition mechanism of N-acetylglycine were elucidated. |
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ISSN: | 0141-0229 1879-0909 |
DOI: | 10.1016/j.enzmictec.2020.109627 |