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Title: Engineering of formate dehydrogenase for the acceptance of a biomimetic nicotinamide-based cofactor in the reduction of CO2
Author: Teijeiro Seijas, Vanesa
Tutor: Ortega-Carrasco, Elisabeth  
Others: Merino, David  
Keywords: protein engineering
nicotinamide biomimetic cofactor
formate dehydrogenase
Issue Date: Jan-2020
Publisher: Universitat Oberta de Catalunya (UOC)
Abstract: Formate dehydrogenase (FDH) is an enzyme widely used in the industry for the regeneration of the cofactor NAD(P)H. Due to the expensive price of the natural cofactor, its regeneration is critical when working with enzymes that require it. Another use of formate dehydrogenase is the reduction of CO2 to formic acid. Biomimetic cofactors were created as an alternative to the expensive natural cofactor NADH. However, there are no studies that integrate them with the enzyme FDH. In this study, using molecular docking, the biomimetic cofactor P3NAH was selected as the biomimetic cofactor that accommodates best in the active site of the enzyme FDH from Pseudomonas (PsFDH) and can reduce CO2. With the objective of creating a mutant of PsFDH that accommodates the nicotinamide part of P3NAH in a position similar to the nicotinamide of NADH, residues were switched in the cofactor access tunnel and active site of the enzyme and the structure of the mutants was generated with homology modelling. When changing the residue Ser 380 for Gly, the nicotinamide part of P3NAH locates the closest when compared to the position of NADH in the WT enzyme. The WT enzyme locates P3NAH with an average distance between relevant atoms of P3NAH and relevant residues of 1.2 Å while the mutant 2NADa_S380G accommodates it with an average distance of 0.25 Å. Apart from acceptable docking results for P3NAH, the protein stability and the ligand transport of 2NADa_S380G is acceptable so this mutant is more suitable for reaction with P3NAH than the WT enzyme.
Language: English
Appears in Collections:Trabajos finales de carrera, trabajos de investigación, etc.

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