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Engineering improved ethylene production: Leveraging systems Biology and adaptive laboratory evolution

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

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Authors

Sophie Vaud

NICOLE PEARCY Nicole.Pearcy@nottingham.ac.uk
Senior Research Fellow in Systems

Marko Hanževački

Alexander M.W. Van Hagen

Laudina Safo

Muhammad Ehsaan

Magdalene Jonczyk

Thomas Millat

SEAN CRAIG SEAN.CRAIG1@NOTTINGHAM.AC.UK
Research Associate

James Fothergill

Rajesh Reddy Bommareddy

Pierre-Yves Colin

Paul Dalby

Christof M. Jäger

Jianping Yu

Pin-Ching Maness

Sean Lynch

Carrie Eckert

Alex Conradie



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 E. 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 the EFE enzyme (WT vs mutant). Our findings provide a novel solution to deregulate metabolic bottlenecks in key pathways, which can be readily applied to address other engineering challenges.

Citation

Vaud, S., Pearcy, N., Hanževački, M., Van Hagen, A. M., Abdelrazig, S., Safo, L., …Bryan, S. J. (2021). Engineering improved ethylene production: Leveraging systems Biology and adaptive laboratory evolution. Metabolic Engineering, 67, 308-320. https://doi.org/10.1016/j.ymben.2021.07.001

Journal Article Type Article
Acceptance Date Jul 2, 2021
Online Publication Date Jul 7, 2021
Publication Date 2021-09
Deposit Date Jul 21, 2021
Publicly Available Date Mar 29, 2024
Journal Metabolic Engineering
Print ISSN 1096-7176
Electronic ISSN 1096-7184
Publisher Elsevier BV
Peer Reviewed Peer Reviewed
Volume 67
Pages 308-320
DOI https://doi.org/10.1016/j.ymben.2021.07.001
Keywords Biotechnology; Applied Microbiology and Biotechnology; Bioengineering
Public URL https://nottingham-repository.worktribe.com/output/5764901
Publisher URL https://www.sciencedirect.com/science/article/pii/S1096717621001117