Exploiting genetic diversity and gene synthesis to identify superior nitrogenase NifH protein variants to engineer N₂-fixation in plants.

ABSTRACT: Engineering nitrogen fixation in eukaryotes requires high expression of functional nitrogenase structural proteins, a goal that has not yet been achieved. Here we build a knowledge-based library containing 32 nitrogenase nifH sequences from prokaryotes of diverse ecological niches and metabolic features and combine with rapid screening in tobacco to identify superior NifH variants for plant mitochondria expression. Three NifH variants outperform in tobacco mitochondria and are further tested in yeast. Hydrogenobacter thermophilus (Aquificae) NifH is isolated in large quantities from yeast mitochondria and fulfills NifH protein requirements for efficient N₂ fixation, including electron transfer for substrate reduction, P-cluster maturation, and FeMo-co biosynthesis. H. thermophil

SUBMITTER: Jiang X 

PROVIDER: S-EPMC7782807 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature