Jianguo Yang, Ph.D.
Nitrogen Fixation and Synthetic Biology research group
Associate Professor, Peking University
tel：
E-mail：yangjg@pku.edu.cn
Engineering and Modification of the Nitrogenase System:
1. Using synthetic biology approaches to engineer the nitrogenase system and enable its functional transfer of the nitrogenase system from prokaryotes to eukaryotes, including major crops;
2. Leveraging the biopersity of diazotrophs and nitrogenase systems to screen and design a minimal nitrogen fixation gene cluster, enabling the transfer of the nitrogenase system to dominant rhizosphere-colonizing microbes in crops;
3. Rational design of high-performance rhizobial strains to enhance symbiotic nitrogen fixation efficiency in leguminous crops (soybean, alfalfa, and peanut, etc.);
4. Development of multi-gene coordinated expression systems for eukaryotes and prokaryotes with broad host compatibility.
Nitrogen is a vital nutrient for crop growth and is a primary factor limiting crop yield. Contemporary agricultural production predominantly depends on the extensive application of chemical nitrogen fertilizers to satisfy the nitrogen requirements of crops. However, the overuse of fertilizers has led to significant environmental issues. Engineering diazotrophs in crops by directly transferring the nitrogenase system into crop cells is a critical strategy for reducing the reliance on agricultural nitrogen fertilizers. Using synthetic biology approaches, we systematically engineered the nitrogenase system, achieving several advancements in simplifying its composition and function. These advancements include: (1) the construction of a minimal FeFe nitrogenase system and the elucidation of its cofactor biosynthesis mechanism (Yang et al., PNAS, 2014); (2) demonstrating that the plant organelle electron transport chain can supply reducing power for substrate reduction by nitrogenase, and proposing the scientific concept of coupling photosynthesis and nitrogen fixation via the electron transport chain (Yang et al., PNAS, 2017); (3) the creation of a polyprotein-based nitrogenase system, significantly simplifying the nitrogenase apparatus (Yang et al., PNAS, 2018; Yang et al., PNAS, 2023); (4) elucidating the molecular mechanism underlying the instability of the core nitrogenase components in eukaryotic mitochondria and identifying ultra-stable core enzyme components (Xiang et al., PNAS, 2020); and (5) developing a multi-gene coordinated expression system with broad host compatibility, providing an essential tool platform for the heterologous expression of the nitrogenase system (Liu et al., Nucleic Acids Res, 2025; Chen et al., Nat Commun, 2026). These achievements have established a robust theoretical and technical foundation for transferring the nitrogenase system into heterologous chassis.
Xie ZT, Cai SY, Chen HY, Kong SY, Tang LT, Wang YP, Yang JG. (2025) Genetic analysis of the NifM dependence of the nitrogenase iron proteins. mBio. e02642-25.
Liu YH, Cai SY, Zhang ZY, Xie ZT, Guo CY, Wang YP, Yang JG. (2025) Expanding the σ54-dependent transcription process with orthogonal designs. Nucleic Acids Res., 53: gkaf442.
Yang JG, Xiang N, Liu YH, Guo CY, Li CY, Li H, Cai SY, Dixon R, Wang YP. (2023) Organelle-dependent polyprotein designs enable stoichiometric expression of nitrogen fixation components targeted to mitochondria. PNAS, 120: e2305142120.
Xiang N, Guo CY, Liu JW, Xu H, Dixon R, Yang JG, Wang YP. (2020) Using synthetic biology to overcome barriers to stable expression of nitrogenase in eukaryotic organelles. PNAS, 117: 16537-16545.
Shuyi Cai, Ziyi Zhang, Yuejie Chen, Zhuoting Xie, Lin Duan, Haoyang Chen, Shuyuan Kong