Cell-free prototyping enables implementation of optimized reverse β-oxidation pathways in heterotrophic and autotrophic bacteria

Bastian Vögeli, Luca Schulz, Shivani Garg, Katia Tarasava, James M. Clomburg, Seung Hwan Lee, Aislinn Gonnot, Elamar Hakim Moully, Blaise R. Kimmel, Loan Tran, Hunter Zeleznik, Steven D. Brown, Sean D. Simpson, Milan Mrksich, Ashty S. Karim, Ramon Gonzalez (), Michael Köpke () and Michael C. Jewett ()
Additional contact information
Bastian Vögeli: Northwestern University
Luca Schulz: Northwestern University
Shivani Garg: LanzaTech Inc.
Katia Tarasava: University of South Florida
James M. Clomburg: LanzaTech Inc.
Seung Hwan Lee: University of South Florida
Aislinn Gonnot: LanzaTech Inc.
Elamar Hakim Moully: Northwestern University
Blaise R. Kimmel: Northwestern University
Loan Tran: LanzaTech Inc.
Hunter Zeleznik: LanzaTech Inc.
Steven D. Brown: LanzaTech Inc.
Sean D. Simpson: LanzaTech Inc.
Milan Mrksich: Northwestern University
Ashty S. Karim: Northwestern University
Ramon Gonzalez: University of South Florida
Michael Köpke: LanzaTech Inc.
Michael C. Jewett: Northwestern University

Nature Communications, 2022, vol. 13, issue 1, 1-10

Abstract: Abstract Carbon-negative synthesis of biochemical products has the potential to mitigate global CO2 emissions. An attractive route to do this is the reverse β-oxidation (r-BOX) pathway coupled to the Wood-Ljungdahl pathway. Here, we optimize and implement r-BOX for the synthesis of C4-C6 acids and alcohols. With a high-throughput in vitro prototyping workflow, we screen 762 unique pathway combinations using cell-free extracts tailored for r-BOX to identify enzyme sets for enhanced product selectivity. Implementation of these pathways into Escherichia coli generates designer strains for the selective production of butanoic acid (4.9 ± 0.1 gL−1), as well as hexanoic acid (3.06 ± 0.03 gL−1) and 1-hexanol (1.0 ± 0.1 gL−1) at the best performance reported to date in this bacterium. We also generate Clostridium autoethanogenum strains able to produce 1-hexanol from syngas, achieving a titer of 0.26 gL−1 in a 1.5 L continuous fermentation. Our strategy enables optimization of r-BOX derived products for biomanufacturing and industrial biotechnology.

Date: 2022
References: View references in EconPapers View complete reference list from CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-022-30571-6 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-30571-6

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-022-30571-6

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().