Functional reconstitution of a bacterial CO2 concentrating mechanism in Escherichia coli

Elife. 2020 Oct 21:9:e59882. doi: 10.7554/eLife.59882.

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 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.

Keywords: E. coli; biochemistry; carboxysome; chemical biology; co2 concentrating mechanism; co2 fixation; infectious disease; microbiology; photosynthesis; synthetic biology.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Carbon Dioxide / metabolism*
  • Escherichia coli / metabolism*
  • Gene Expression Regulation, Bacterial / physiology*
  • Genome, Bacterial
  • Genomics
  • Halothiobacillus / genetics
  • Mutation
  • Phosphotransferases (Alcohol Group Acceptor) / genetics
  • Phosphotransferases (Alcohol Group Acceptor) / metabolism
  • Ribulose-Bisphosphate Carboxylase / genetics
  • Ribulose-Bisphosphate Carboxylase / metabolism

Substances

  • Bacterial Proteins
  • Carbon Dioxide
  • Phosphotransferases (Alcohol Group Acceptor)
  • phosphoribulokinase
  • Ribulose-Bisphosphate Carboxylase