Engineering Improved Ethylene Production: Leveraging Systems Biology and Adaptive Laboratory Evolution

Sophie Vaud, Nicole Pearcy, Marko Hanževacki, Alexander Van Hagen, Salah Abdelrazig, Laudina Safo, Muhammad Ehsaan, Magdalene Jonczyk, Thomas Millat, Sean Craig, Edward Spence, James Fothergill, Rajesh Bommareddy, Pierre-Yves Colin, Jamie Twycross, Paul Dalby, Nigel Minton, Christof Jäger, Dong-Hyun Kim, Jianping YuPin-Ching Maness, Sean Lynch, Carrie Eckert, Alex Conradie, Samantha Bryan

Research output: Contribution to journalArticlepeer-review

6 Scopus Citations

Abstract

Ethylene is a small hydrocarbon gas widely used in the chemical industry. Annual worldwide production currently exceeds 150 million tons, producing considerable amounts of CO2 contributing to climate change. The need for a sustainable alternative is therefore imperative. Ethylene is natively produced by several different microorganisms, including Pseudomonas syringae pv. phaseolicola via a process catalyzed by the ethylene-forming enzyme (EFE), subsequent heterologous expression of EFE has led to ethylene production in non-native bacterial hosts including Escherichia coli and cyanobacteria. However, solubility of EFE and substrate availability remain rate-limiting steps in biological ethylene production. We employed a combination of genome-scale metabolic modelling, continuous fermentation, and protein evolution to enable the accelerated development of a high efficiency ethylene producing E. coli strain, yielding a 49-fold increase in production, the most significant improvement reported to date. Furthermore, we have clearly demonstrated that this increased yield resulted from metabolic adaptations that were uniquely linked to EFE (wild type versus mutant). Our findings provide a novel solution to deregulate metabolic bottlenecks in key pathways, which can be readily applied to address other engineering challenges.

Original languageAmerican English
Pages (from-to)308-320
Number of pages13
JournalMetabolic Engineering
Volume67
DOIs
StatePublished - Sep 2021

Bibliographical note

Publisher Copyright:
© 2021 The Authors

NREL Publication Number

  • NREL/JA-2700-79652

Keywords

  • Adaptive evolution
  • Directed evolution
  • Fermentation
  • Metabolic engineering
  • Systems biology

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