2-Ketogluconate Kinase from Cupriavidus necator H16: Purification, Characterization, and Exploration of Its Substrate Specificity

Israel Sanchez-Moreno (1) , Natalia Trachtmann (2) , Sibel Ilhan (1, 2) , Virgil Hélaine (1) , Marielle Lemaire (1) , Christine Guérard-Hélaine (1) , Georg Sprenger (2)

1 ICCF - Institut de Chimie de Clermont-Ferrand
2 Universität Stuttgart [Stuttgart]

We have cloned, overexpressed, purified, and characterized a 2-ketogluconate kinase (2-dehydrogluconokinase, EC 2.7.1.13) from Cupriavidus necator (Ralstonia eutropha) H16. Exploration of its substrate specificity revealed that three ketoacids (2-keto-3-deoxy-d-gluconate, 2-keto-d-gulonate, and 2-keto-3-deoxy-d-gulonate) with structures close to the natural substrate (2-keto-d-gluconate) were successfully phosphorylated at an efficiency lower than or comparable to 2-ketogluconate, as depicted by the measured kinetic constant values. Eleven aldo and keto monosaccharides of different chain lengths and stereochemistries were also assayed but not found to be substrates. 2-ketogluconate-6-phosphate was synthesized at a preparative scale and was fully characterized for the first time.

Mots clés: 2-ketogluconate; 2-ketogluconate kinase; 2-ketogulonate; 2-keto-3-deoxygluconate; Cupriavidus necator; biocatalysis; monosaccharides phosphate

Domaines: Chimie organique; Catalyse; Biotechnologies

Israel Sanchez-Moreno, Natalia Trachtmann, Sibel Ilhan, Virgil Hélaine, Marielle Lemaire, et al.. 2-Ketogluconate Kinase from Cupriavidus necator H16: Purification, Characterization, and Exploration of Its Substrate Specificity. Molecules, 2019, 24 (13), pp.2393. ⟨10.3390/molecules24132393⟩. ⟨hal-02414422⟩