Nitrogenase cofactor biosynthesis using proteins produced in mitochondria of <i>Saccharomyces cerevisiae</i>
Katarzyna Dobrzyńska, Ana Pérez‐González, Carlos Echávarri‐Erasun, Diana Coroian, Alvaro Salinero‐Lanzarote, Marcel Veldhuizen, Dennis R. Dean, Stefan Burén, Luis M. Rubio
Abstract
Biological nitrogen fixation, the conversion of inert N2 to metabolically usable NH3, is a process exclusive to diazotrophic microorganisms and relies on the activity of nitrogenases. The assembly of the nitrogenase [7Fe-9S-C-Mo- R -homocitrate]-cofactor (FeMo-co) in a eukaryotic cell is a pivotal milestone that will pave the way to engineer cereals with nitrogen fixing capabilities and therefore independent of nitrogen fertilizers. In this study, we identified NifEN protein complexes that were functional in the model eukaryotic organism Saccharomyces cerevisiae . NifEN is an essential component of the FeMo-co biosynthesis pathway. Furthermore, the FeMo-co biosynthetic pathway was recapitulated in vitro using only proteins expressed in S. cerevisiae . FeMo-co biosynthesis was achieved by combining nitrogenase FeMo-co assembly components from different species, a promising strategy to engineer nitrogen fixation in eukaryotic organisms.