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Cell-free prototyping enables implementation of optimized reverse β-oxidation pathways in heterotrophic and autotrophic bacteria.
- Source :
-
Nature communications [Nat Commun] 2022 Jun 01; Vol. 13 (1), pp. 3058. Date of Electronic Publication: 2022 Jun 01. - Publication Year :
- 2022
-
Abstract
- Carbon-negative synthesis of biochemical products has the potential to mitigate global CO <subscript>2</subscript> 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 <superscript>-1</superscript> ), as well as hexanoic acid (3.06 ± 0.03 gL <superscript>-1</superscript> ) and 1-hexanol (1.0 ± 0.1 gL <superscript>-1</superscript> ) 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 <superscript>-1</superscript> in a 1.5 L continuous fermentation. Our strategy enables optimization of r-BOX derived products for biomanufacturing and industrial biotechnology.<br /> (© 2022. The Author(s).)
Details
- Language :
- English
- ISSN :
- 2041-1723
- Volume :
- 13
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nature communications
- Publication Type :
- Academic Journal
- Accession number :
- 35650184
- Full Text :
- https://doi.org/10.1038/s41467-022-30571-6