@article{827825d38cc944febb152f1eded64e52,
title = "Functional reconstitution of a bacterial CO 2 concentrating mechanism in Escherichia coli.",
abstract = "Many photosynthetic organisms employ a CO2 concentrating mechanism (CCM) to increase the rate of CO2 fixation via the Calvin cycle. CCMs catalyze approximate to 50% of global photosynthesis, yet it remains unclear which genes and proteins are required to produce this complex adaptation. We describe the construction of a functional CCM in a non-native host, achieved by expressing genes from an autotrophic bacterium in an Escherichia coli strain engineered to depend on rubisco carboxylation for growth. Expression of 20 CCM genes enabled E. coli to grow by fixing CO2 from ambient air into biomass, with growth in ambient air depending on the components of the CCM. Bacterial CCMs are therefore genetically compact and readily transplanted, rationalizing their presence in diverse bacteria. Reconstitution enabled genetic experiments refining our understanding of the CCM, thereby laying the groundwork for deeper study and engineering of the cell biology supporting CO2 assimilation in diverse organisms.",
author = "Flamholz, {Avi I.} and Eli Dugan and Cecilia Blikstad and Shmuel Gleizer and Roee Ben-Nissan and Shira Amram and Niv Antonovsky and Sumedha Ravishankar and Elad Noor and Arren Bar-Even and Ron Milo and Savage, {David F.}",
note = "This work was supported by a National Science Foundation Graduate Research Fellowship (to A.I.F.), the US Department of Energy (grant DE-SC00016240) and Royal Dutch Shell (Energy Biosciences Institute project CW163755) for CCM characterization (to D.F.S.), the National Science Foundation (grant MCB-1818377) for CCMB1 construction (to D.F.S.), and by the European Research Council (Project NOVCARBFIX 646827) to R.M. R.M. is the Charles and Louise Gartner Professional Chair. We dedicate this paper to the memory of Arren Bar-Even, who was a great friend and teacher and whose wit and intellect inspired us throughout this work. Thanks to Matt Davis for P1 transduction materials and advice, Hernan Garcia and Han Lim for pZ plasmids, Maggie Stoeva, Anna Engelbrektson, Anchal Mehra, Sophia Ewens and Tyler Barnum for help with anaerobic cultivation, Reena Zalpuri and Danielle Jorgens at the University of California Berkeley Electron Microscope Laboratory for advice and assistance with electron microscopy, and Rob Egbert and Adam Arkin for KEIO strains. We are grateful to Griffin Chure, Eric Estrin, Woody Fischer, Evan Groober, Darcy McRose, Dipti Nayak, Sabeeha Merchant, Luke Oltrogge, Naiya Phillips, and Ari Satanowski for detailed comments on the manuscript, and to Dan Arlow, Yinon Bar-On, Dan Davidi, Jack Desmarais, Hernan Garcia, Oliver Mueller-Cajar, Rob Nichols, Kris Niyogi, Dan Portnoy, Morgan Price, Noam Prywes, Jeremy Roop, Rachel Shipps, Patrick Shih, and Dan Tawfik, for support, advice and helpful discussions throughout. Avi I Flamholz, Conceptualization, Resources, Data curation, Software, Formal analysis, Funding acquisition, Validation, Investigation, Visualization, Methodology, Writing - original draft, Project administration, Writing - review and editing; Eli Dugan, Roee Ben-Nissan, Validation, Investigation, Methodology, Writing - review and editing; Cecilia Blikstad, Data curation, Formal analysis, Investigation, Methodology, Writing review and editing; Shmuel Gleizer, Data curation, Formal analysis, Validation, Investigation, Methodology, Writing - review and editing; Shira Amram, Resources, Investigation, Methodology; Niv Antonovsky, Conceptualization, Resources, Investigation, Methodology, Writing - review and editing; Sumedha Ravishankar, Investigation, Methodology; Elad Noor, Conceptualization, Software, Formal analysis, Investigation, Methodology, Writing - review and editing; Arren Bar-Even, Conceptualization, Resources, Supervision, Methodology, Writing - review and editing; Ron Milo, David F Savage, Conceptualization, Resources, Supervision, Funding acquisition, Methodology, Project administration, Writing - review and editin. U.S. Department of Energy (DE-SC00016240), David F Savage, European Research Council (NOVCARBFIX 646827), Ron Milo, National Science Foundation (MCB-1818377), David F Savage. Shell (EBI CW163755), David F Savage, The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.",
year = "2020",
month = oct,
day = "21",
doi = "https://doi.org/10.7554/eLife.59882",
language = "الإنجليزيّة",
volume = "9",
pages = "1--57",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",
}