Production and characterization of CO2 reducing and tungsten-containing formate dehydrogenase of Cupriavidus necator H16

• Published in: Biotechnology and bioprocess engineering Vol. 30; no. 2; pp. 363 - 376
• Main Authors: Ryu, Huichang, Nguyen, Ngoc Minh Chau, Park, Hyung-Yeon, Lee, Sung Kuk, Park, Sunghoon
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Nature B.V 01.04.2025; 한국생물공학회
• Subjects: Carbon dioxide; Carbon dioxide emissions; Cloning; Cupriavidus necator; Dehydrogenase; Dehydrogenases; Emissions control; Enzymes; Formate dehydrogenase; formates; Genes; guanine; oxygen; Plasmids; Proteins; Steric hindrance; Tungsten; 생물공학
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-024-00171-w

Formate dehydrogenases, such as MeFDH1 from Methylorubrum extorquens, efficiently convert CO2 to formate using the tungsto-bis-metallopterin guanine dinucleotide cofactor, aiding in CO2 emission reduction. This study characterizes a similar enzyme, tungsten-containing formate dehydrogenase from Cupriavidus necator H16, expressed in M. extorquens. CnFDW, with α and β subunits, shows high formate-forming activity (kcat = 120 s⁻1) using reduced ethyl viologen as an electron donor. Tungstate presence is crucial for activity, although inactive apo-CnFDW is synthesized without it. HR-ICP-MS analysis revealed a maximal tungsten content of 0.56 mol/mol enzyme, indicating 44% of enzymes lack the cofactor. CnFDW exhibited optimal formate production at 30 °C and pH 6.0, with high oxygen tolerance in the absence of formate. Docking studies suggest steric hindrance in cofactor binding reduces its activity and W content compared to MeFDH1, positioning CnFDW as a promising candidate for CO2-to-formate conversion.