Expressing 2-keto acid pathway enzymes significantly increases photosynthetic isobutanol production
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Abstract Background Cyanobacteria, photosynthetic microorganisms, are promising green cell factories for chemical production, including biofuels. Isobutanol, a four-carbon alcohol, is considered as a superior candidate as a biofuel for its high energy density with suitable chemical and physical characteristics. The unicellular cyanobacteriumSynechocystisPCC 6803 has been successfully engineered for photosynthetic isobutanol production from CO2and solar energy in a direct process. Results Heterologous expression of α-ketoisovalerate decarboxylase (KivdS286T) is sufficient for isobutanol synthesis via the 2-keto acid pathway inSynechocystis. With additional expression of acetolactate synthase (AlsS), acetohydroxy-acid isomeroreductase (IlvC), dihydroxy-acid dehydratase (IlvD), and alcohol dehydrogenase (Slr1192OP), theSynechocystisstrain HX42, with a functional 2-keto acid pathway, showed enhanced isobutanol production reaching 98 mg L−1in short-term screening experiments. Through modulatingkivdS286Tcopy numbers as well as the composition of the 5′-region, a finalSynechocystisstrain HX47 with three copies ofkivdS286Tshowed a significantly improved isobutanol production of 144 mg L−1, an 177% increase compared to the previously reported best producing strain under identical conditions. Conclusions This work demonstrates the feasibility to express heterologous genes with a combination of self-replicating plasmid-based system and genome-based system inSynechocystiscells. Obtained isobutanol-producingSynechocystisstrains form the base for further investigation of continuous, long-term-photosynthetic isobutanol production from solar energy and carbon dioxide. Graphic abstract
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Ethanol and isobutanol are mainly derived from pyruvate-based pathways.
“ethanol and isobutanol are mainly derived from pyruvate-based pathways [27]”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [27]
The Introduction states that the isobutanol-producing 2-keto acid pathway starts with pyruvate, and the Results and discussion identifies pyruvate as its precursor. Our copy does not establish the corresponding claim for ethanol or that these are the main pathways for both products. (LLM, conf=0.88)
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- Authors
- Xie, H., & Lindblad, P.
- Affiliations
- Uppsala University (SE)
- Corresponding author
- Lindblad, Peter
- Venue
- Microbial Cell Factories
- Published
- 2022-02-01
- Volume
- 21
- Issue
- 1
- Pages
- 17-17
- ISSN
- 1475-2859 (ISSN-L)
- PMC
- PMC8805274
- OpenAlex
- W4210723962
- Cited by
- 17
- References
- 39
- Citations by year
- 2026: 4 · 2025: 6 · 2024: 2 · 2023: 4 · 2022: 1
- FWCI
- 1.06
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Microbial Cell Factories (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Uppsala University Publications (Uppsala University) (submittedVersion, other-oa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Energimyndigheten (P46607-1)
- NordForsk (82845)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant biochemistry and biosynthesis
- Keywords
- Synechocystis sp. PCC 6803, cyanobacteria, acetolactate synthase, alcohol dehydrogenase, heterologous expression, plasmid copy number, biofuels, CO2 fixation, solar energy
- MeSH terms
- Butanols, Bacterial Proteins, Carbon Dioxide, Keto Acids, Photosynthesis, Synechocystis, Biosynthetic Pathways, Metabolic Engineering
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