Biochemical and Kinetic Characterization of the Glucose-6-Phosphate Dehydrogenase from Helicobacter pylori Strain 29CaP

• Published in: Microorganisms (Basel) 2022-07, Vol.10 (7), p.1359
• Main Authors: Ortiz-Ramírez, Paulina, Hernández-Ochoa, Beatriz, Ortega-Cuellar, Daniel, González-Valdez, Abigail, Martínez-Rosas, Víctor, Morales-Luna, Laura, Arreguin-Espinosa, Roberto, Castillo-Rodríguez, Rosa Angélica, Canseco-Ávila, Luis Miguel, Cárdenas-Rodríguez, Noemi, Pérez de la Cruz, Verónica, Montiel-González, Alba Mónica, Gómez-Chávez, Fernando, Gómez-Manzo, Saúl
• Format: Article
• Language: English
• Subjects: Antibiotics; biochemical characterization; Biochemistry; Cancer therapies; Chromatography; Cloning; Dehydrogenase; Dehydrogenases; E coli; Enzymatic activity; Enzyme activity; Enzymes; Equilibrium; Gastric cancer; Glucose; Glucose 6 phosphate dehydrogenase; Glucose metabolism; Glucosephosphate dehydrogenase; Glycerol; Guanidine hydrochloride; Helicobacter pylori; Infections; Kinases; kinetic; Metabolism; Optimization; Pathogens; Pentose; Pentose phosphate pathway; Phosphates; Plasmids; Proteins; Risk analysis; Risk factors; Thermal stability; Three dimensional models; Trypsin
• ISSN: 2076-2607; 2076-2607
• DOI: 10.3390/microorganisms10071359

Helicobacter pylori (H. pylori) has been proposed as the foremost risk factor for the development of gastric cancer. We found that H. pylori express the enzyme glucose-6-phosphate dehydrogenase (HpG6PD), which participates in glucose metabolism via the pentose phosphate pathway. Thus, we hypothesized that if the biochemical and physicochemical characteristics of HpG6PD contrast with the host G6PD (human G6PD, HsG6PD), HpG6PD becomes a potential target for novel drugs against H. pylori. In this work, we characterized the biochemical properties of the HpG6PD from the H.pylori strain 29CaP and expressed the active recombinant protein, to analyze its steady-state kinetics, thermostability, and biophysical aspects. In addition, we analyzed the HpG6PD in silico structural properties to compare them with those of the HsG6PD. The optimal pH for enzyme activity was 7.5, with a T1/2 of 46.6 °C, at an optimum stability temperature of 37 °C. The apparent Km values calculated for G6P and NADP+ were 75.0 and 12.8 µM, respectively. G6P does not protect HpG6PD from trypsin digestion, but NADP+ does protect the enzyme from trypsin and guanidine hydrochloride (Gdn-HCl). The biochemical characterization of HpG6PD contributes to knowledge regarding H. pylori metabolism and opens up the possibility of using this enzyme as a potential target for specific and efficient treatment against this pathogen; structural alignment indicates that the three-dimensional (3D) homodimer model of the G6PD protein from H. pylori is different from the 3D G6PD of Homo sapiens.