Electro-enzymatic ATP regeneration coupled to biocatalytic phosphorylation reactions

• Published in: Bioelectrochemistry (Amsterdam, Netherlands) Vol. 152; p. 108432
• Main Authors: García-Molina, Gabriel, Natale, Paolo, Coito, Ana M., Cava, Daniel G., A. C. Pereira, Inés, López-Montero, Iván, Vélez, Marisela, Pita, Marcos, De Lacey, Antonio L.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-08-2023
• Subjects: ATP regeneration; ATP-synthase; Hydrogenase; Kinase; Phosphorylation; Hydrogenase; ATP regeneration; Phosphorylation; ATP-synthase; Kinase
• ISSN: 1567-5394; 1878-562X
• DOI: 10.1016/j.bioelechem.2023.108432

•Hydrogenase and ATPase reconstituted in a biomimetic membrane over an electrode.•Electro-enzymatic ATP regeneration for phosphorylation of glucose is achieved.•Electro-enzymatic ATP regeneration for synthesis of NADP+ is achieved.
Adenosine-5-triphosphate (ATP) is the main energy vector in biological systems, thus its regeneration is an important issue for the application of many enzymes of interest in biocatalysis and synthetic biology. We have developed an electroenzymatic ATP regeneration system consisting in a gold electrode modified with a floating phospholipid bilayer that allows coupling the catalytic activity of two membrane-bound enzymes: NiFeSe hydrogenase from Desulfovibrio vulgaris and F1Fo-ATP synthase from Escherichia coli. Thus, H2 is used as a fuel for producing ATP. This electro-enzymatic assembly is studied as ATP regeneration system of phosphorylation reactions catalysed by kinases, such as hexokinase and NAD+-kinase for respectively producing glucose-6-phosphate and NADP+.