Abstract
Synthetic autotrophy is a promising avenue to sustainable bioproduction from CO2. Here, we use iterative laboratory evolution to generate several distinct autotrophic strains. Utilising this genetic diversity, we identify that just three mutations are sufficient for Escherichia coli to grow autotrophically, when introduced alongside non-native energy (formate dehydrogenase) and carbon-fixing (RuBisCO, phosphoribulokinase, carbonic anhydrase) modules. The mutated genes are involved in glycolysis (pgi), central-carbon regulation (crp), and RNA transcription (rpoB). The pgi mutation reduces the enzyme's activity, thereby stabilising the carbon-fixing cycle by capping a major branching flux. For the other two mutations, we observe down-regulation of several metabolic pathways and increased expression of native genes associated with the carbon-fixing module (rpiB) and the energy module (fdoGH), as well as an increased ratio of NADH/NAD+ - the cycle's electron-donor. This study demonstrates the malleability of metabolism and its capacity to switch trophic modes using only a small number of genetic changes and could facilitate transforming other heterotrophic organisms into autotrophs.
Original language | English |
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Article number | RP88793 |
Number of pages | 22 |
Journal | eLife |
Volume | 12 |
DOIs | |
Publication status | Published - 21 Feb 2024 |
Funding
We thank Alon Barshap, Alon Savidor, Aliza Fedorenko, Avi Flamholz, Avihu Yona, Daria Fedorova, Emanuel Avrahami, Ifat, Goldstein, Ilana Rogachev, Ido Cohen, Lior Greenspoon, Lior Shachar, Margarita Gortikov, Merav Hagag, Niv Antonovsky, Ofir Shechter, Ron Sender, Samuel Lovat, Tasneem Bareia, Tali Wiesel, Yafit Sugas, Yinon Bar-on, Yotam David, Yuval Kushmaro and Yuval Rosenberg for their support of this project. We acknowledge the De Botton Protein Profiling institute of the Nancy and Stephen Grand Israel National Center for Personalized Medicine, Weizmann Institute of Science for their services and contribution to this project. This research was generously supported by the Mary and Tom Beck Canadian Center for Alternative Energy Research, the Schwartz-Reisman Collaborative Science Program, the Ullmann Family Foundation and the Yotam Project. Prof. Ron Milo is the Head of the Mary and Tom Beck Canadian Center for Alternative Energy Research and the incumbent of the Charles and Louise Gartner Professorial Chair. RBN is a Weizmann SAERI fellow. EM is a fellow of the Ariane de Rothschild Women Doctoral Program. HL and VP acknowledge funding from the Cluster of Excellence EXC 2124 from the Deutsche Forschungsgemeinschaft. Funding: Mary and Tom Beck Canadian Center for Alternative Energy Research (RM), Schwartz-Reisman Collaborative Science Program (RM), Ullmann Family Foundation and the Yotam Project (RM), Sustainability and Energy Weizmann Doctoral Fellowship (RBN), Ariane de Rothschild Women Doctoral Program (EM), Cluster of Excellence EXC 2124 from the Deutsche Forschungsgemeinschaft (HL, VP) Publisher Copyright: © 2023, Ben Nissan, Milshtein et al.
All Science Journal Classification (ASJC) codes
- General Neuroscience
- General Biochemistry,Genetics and Molecular Biology
- General Immunology and Microbiology