Functional reconstitution of a bacterial CO2 concentrating mechanism in E. coli

Flamholz, A. I.; Dugan, E.; Blikstad, C.; Gleizer, S.; Ben-Nissan, R.; Amram, S.; Antonovsky, N.; Ravishankar, S.; Noor, E.; Bar-Even, A.; Milo, R.; Savage, D.

doi:10.1101/2020.05.27.119784

bioRxiv · 10.1101/2020.05.27.119784

Functional reconstitution of a bacterial CO2 concentrating mechanism in E. coli

Flamholz, A. I.·Dugan, E.·Blikstad, C.·Gleizer, S.·Ben-Nissan, R.·Amram, S.·Antonovsky, N.·Ravishankar, S.·Noor, E.·Bar-Even, A.·Milo, R.·Savage, D.

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Abstract

Many photosynthetic organisms employ a CO2 concentrating mechanism (CCM) to increase the rate of CO2 fixation via the Calvin cycle. CCMs catalyze ≈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 engineered E. coli strain. Expression of 20 CCM genes enabled E. coli to grow by fixing CO2 from ambient air into biomass, with growth depending on CCM components. 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.One Sentence Summary A bacterial CO2 concentrating mechanism enables E. coli to fix CO2 from ambient air.Competing Interest StatementD.F.S. is a co-founder of Scribe Therapeutics and a scientific advisory board member of Scribe Therapeutics and Mammoth Biosciences. A.B.-E. is co-founder of b.fab. These companies were not involved in this research in any way. All other authors declare no competing interests.View Full Text

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Flamholz, A. I., Dugan, E., Blikstad, C., Gleizer, S., Ben-Nissan, R., Amram, S., Antonovsky, N., Ravishankar, S., Noor, E., Bar-Even, A., Milo, R., Savage, D.. 2020-05-27. Functional reconstitution of a bacterial CO2 concentrating mechanism in E. coli. https://doi.org/10.1101/2020.05.27.119784

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