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Organelle-dependent polyprotein designs enable stoichiometric expression of nitrogen fixation components targeted to mitochondria

/ 2023-08-17
AuthorJianguo Yang , Nan Xiang , Yiheng Liu, Chenyue Guo, Chenyu Li, Hui Li, Shuyi Cai, ,and Yi-Ping Wang 
Paper journal:PNAS
Engineering the capacity for biological nitrogen fixation in cereals would have tremendous environmental and socioeconomic benefits, since this process provides a more sustainable alternative to the application of chemical fertilizers. The physiological environment and compartmentalized structure of mitochondria provide a unique location for metabolic pathway reconstitution, including nitrogen fixation. In the current study, we redesigned a polyprotein system linked by engineered peptides cleaved by the intrinsic mitochondrial-processing peptidase that enables stable expression of protein components required for nitrogenase biosynthesis in yeast mitochondria. In addition to engineering nitrogen fixation, we demonstrate that this polyprotein approach expands the potential for utilizing mitochondria as a biotechnology platform through functional reconstitution of the violacein and isobutanol pathways in this organelle.