Introduction of NADH-dependent nitrate assimilation in Synechococcus sp. PCC 7002 improves photosynthetic production of 2-methyl-1-butanol and isobutanol
Open accessAbstract
Cyanobacteria hold promise for renewable chemical production due to their photosynthetic nature, but engineered strains frequently display poor production characteristics. These difficulties likely arise in part due to the distinctive photoautotrophic metabolism of cyanobacteria. In this work, we apply a genome-scale metabolic model of the cyanobacteria Synechococus sp. PCC 7002 to identify strain designs accounting for this unique metabolism that are predicted to improve the production of various biofuel alcohols (e.g. 2-methyl-1-butanol, isobutanol, and 1-butanol) synthesized via an engineered biosynthesis pathway. Using the model, we identify that the introduction of a large, non-native NADH-demand into PCC 7002's metabolic network is predicted to enhance production of these alcohols by promoting NADH-generating reactions upstream of the production pathways. To test this, we construct strains of PCC 7002 that utilize a heterologous, NADH-dependent nitrite reductase in place of the native, ferredoxin-dependent enzyme to create an NADH-demand in the cells when grown on nitrate-containing media. We find that photosynthetic production of both isobutanol and 2-methyl-1-butanol is significantly improved in the engineered strain background relative to that in a wild-type background. We additionally identify that the use of high-nutrient media leads to a substantial prolongment of the production curve in our alcohol production strains. The metabolic engineering strategy identified and tested in this work presents a novel approach to engineer cyanobacterial production strains that takes advantage of a unique aspect of their metabolism and serves as a basis on which to further develop strains with improved production of these alcohols and related products.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Under optimal laboratory conditions, Synechococcus sp. PCC 7002 can attain higher biomass productivity than several commonly used cyanobacterial hosts.
“Under optimal laboratory conditions, this strain can attain higher biomass productivity than several commonly used cyanobacterial hosts, thereby enhancing its potential for industrial biotechnology applications [64,66].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [66]
The Introduction (page unavailable) describes PCC 7002 as fast-growing and robust, and says it can grow at temperatures and salinities beyond those tolerated by most other model cyanobacteria. It does not establish that PCC 7002 has higher biomass productivity than several commonly used hosts under optimal laboratory conditions. (LLM, conf=0.82)
Experimental — accuracy not yet measuredSynechococcus sp. PCC 7002 has a comparatively rapid growth rate compared with other cyanobacteria used in metabolic engineering.
“A significant advantage of Synechococcus sp. PCC 7002 is its comparatively rapid growth rate compared with other cyanobacteria used in metabolic engineering.”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [66]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels
Atsumi, S., Hanai, T., & Liao, J. C. (2008) · Nature · doi:10.1038/nature06450 · Cited by 1,945 · see citing works
Burgard, A. P., Pharkya, P., & Maranas, C. D. (2003) · Biotechnology and Bioengineering · doi:10.1002/bit.10803 · Cited by 1,262 · see citing works
Direct photosynthetic recycling of carbon dioxide to isobutyraldehyde
Atsumi, S., Higashide, W., & Liao, J. C. (2009) · Nature Biotechnology · doi:10.1038/nbt.1586 · Cited by 870 · see citing works
Using Genome-scale Models to Predict Biological Capabilities
O’Brien, E. J., Monk, J. M., & Palsson, B. Ø. (2015) · Cell · doi:10.1016/j.cell.2015.05.019 · Cited by 709 · see citing works
Moreno‐Vivián, C., Cabello, P., Martínez‐Luque, M., Blasco, R., & del Castillo, F. J. (1999) · Journal of Bacteriology · doi:10.1128/jb.181.21.6573-6584.1999 · Cited by 593 · see citing works
Applications of cyanobacteria in biotechnology
Abed, R. M. M., Dobretsov, S., & Sudesh, K. (2009) · Journal of Applied Microbiology · doi:10.1111/j.1365-2672.2008.03918.x · Cited by 528 · see citing works
Metabolic engineering of cyanobacteria for 1-butanol production from carbon dioxide
Lan, E. I., & Liao, J. C. (2011) · Metabolic Engineering · doi:10.1016/j.ymben.2011.04.004 · Cited by 382 · see citing works
NDH-1 and NDH-2 Plastoquinone Reductases in Oxygenic Photosynthesis
Peltier, G., Aro, E., & Shikanai, T. (2016) · Annual Review of Plant Biology · doi:10.1146/annurev-arplant-043014-114752 · Cited by 304 · see citing works
Detailing the optimality of photosynthesis in cyanobacteria through systems biology analysis
Nogales, J., Gudmundsson, S., Knight, E. M., Palsson, B. O., & Thiele, I. (2012) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1117907109 · Cited by 301 · see citing works
Ammonium assimilation in cyanobacteria
Muro-Pastor, M. I., Reyes, J. C., & Florencio, F. J. (2005) · Photosynthesis Research · doi:10.1007/s11120-004-2082-7 · Cited by 287 · see citing works
Davies, F. K., Work, V. H., Beliaev, A. S., & Posewitz, M. C. (2014) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2014.00021 · Cited by 273 · see citing works
Metabolic engineering of cyanobacteria for the synthesis of commodity products
Angermayr, S. A., Gorchs Rovira, A., & Hellingwerf, K. J. (2015) · Trends in Biotechnology · doi:10.1016/j.tibtech.2015.03.009 · Cited by 273 · see citing works
Energy biotechnology with cyanobacteria
Angermayr, S. A., Hellingwerf, K. J., Lindblad, P. C., & de Mattos, M. J. T. (2009) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2009.05.011 · Cited by 255 · see citing works
Atsumi, S., & Liao, J. C. (2008) · Applied and Environmental Microbiology · doi:10.1128/aem.02046-08 · Cited by 249 · see citing works
Knoop, H., Gründel, M., Zilliges, Y., Lehmann, R., Hoffmann, S., Lockau, W., & Steuer, R. (2013) · PLoS Computational Biology · doi:10.1371/journal.pcbi.1003081 · Cited by 248 · see citing works
Cournac, L., Guedeney, G., Peltier, G., & Vignais, P. M. (2004) · Journal of Bacteriology · doi:10.1128/jb.186.6.1737-1746.2003 · Cited by 246 · see citing works
Photosynthetic nitrate assimilation in cyanobacteria
Flores, E., Frías, J. E., Rubio, L. M., & Antonia Herrero, B. (2005) · Photosynthesis Research · doi:10.1007/s11120-004-5830-9 · Cited by 241 · see citing works
Tamoi, M., Miyazaki, T., Fukamizo, T., & Shigeoka, S. (2005) · The Plant Journal · doi:10.1111/j.1365-313x.2005.02391.x · Cited by 236 · see citing works
Synthetic Biology in Cyanobacteria
Heidorn, T., Camsund, D., Huang, H.-H., Lindberg, P., Oliveira, P., Stensjö, K., & Lindblad, P. C. (2011) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/b978-0-12-385075-1.00024-x · Cited by 235 · see citing works
Cooley, J. W., & Vermaas, W. F. J. (2001) · Journal of Bacteriology · doi:10.1128/jb.183.14.4251-4258.2001 · Cited by 214 · see citing works
Markley, A. L., Begemann, M. B., Clarke, R., Gordon, G. C., & Pfleger, B. F. (2014) · ACS Synthetic Biology · doi:10.1021/sb500260k · Cited by 212 · see citing works
Self-citationWłodarczyk, A., Selão, T. T., Norling, B., & Nixon, P. J. (2020) · Communications Biology · doi:10.1038/s42003-020-0910-8 · Cited by 192 · see citing works
Metabolic Engineering of Synechocystis sp. Strain PCC 6803 for Isobutanol Production
Varman, A. M., Xiao, Y., Pakrasi, H. B., & Tang, Y. (2012) · Applied and Environmental Microbiology · doi:10.1128/aem.02827-12 · Cited by 178 · see citing works
Engineering of photosynthetic mannitol biosynthesis from CO2 in a cyanobacterium
Jacobsen, J. H., & Frigaard, N. (2013) · Metabolic Engineering · doi:10.1016/j.ymben.2013.11.004 · Cited by 153 · see citing works
Kusakabe, T., Tatsuke, T., Tsuruno, K., Hirokawa, Y., Atsumi, S., Liao, J. C., & Hanai, T. (2013) · Metabolic Engineering · doi:10.1016/j.ymben.2013.09.007 · Cited by 149 · see citing works
Regulation of nitrate assimilation in cyanobacteria
Ohashi, Y., Shi, W., Takatani, N., Aichi, M., Maeda, S.-I., Watanabe, S., Yoshikawa, H., & Omata, T. (2011) · Journal of Experimental Botany · doi:10.1093/jxb/erq427 · Cited by 149 · see citing works
Bacterial nitrate assimilation: gene distribution and regulation
Luque‐Almagro, V. M., Gates, A. J., Moreno‐Vivián, C., et al. (2011) · Biochemical Society Transactions · doi:10.1042/bst20110688 · Cited by 142 · see citing works
Ludwig, M., & Bryant, D. A. (2011) · Frontiers in Microbiology · doi:10.3389/fmicb.2011.00041 · Cited by 136 · see citing works
NAKANO, M. M., Hoffmann, T., Zhu, Y., & Jahn, D. (1998) · Journal of Bacteriology · doi:10.1128/jb.180.20.5344-5350.1998 · Cited by 136 · see citing works
Metabolic design for cyanobacterial chemical synthesis
Oliver, J. W. K., & Atsumi, S. (2014) · Photosynthesis Research · doi:10.1007/s11120-014-9997-4 · Cited by 134 · see citing works
Broddrick, J. T., Rubin, B. E., Welkie, D. G., Du, N., Mih, N., Diamond, S., Lee, J. J., Golden, S. S., & Palsson, B. Ø. (2016) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1613446113 · Cited by 126 · see citing works
Gutthann, F., Egert, M., Marques, A., & Appel, J. (2006) · Biochimica et Biophysica Acta (BBA) - Bioenergetics · doi:10.1016/j.bbabio.2006.12.003 · Cited by 124 · see citing works
Shen, C. R., & Liao, J. C. (2012) · Energy & Environmental Science · doi:10.1039/c2ee23148d · Cited by 116 · see citing works
Computational methods in metabolic engineering for strain design
Long, M. R., Ong, W. K., & Reed, J. L. (2015) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2014.12.019 · Cited by 110 · see citing works
Self-citationEngineered cyanobacteria: Teaching an old bug new tricks
Ruffing, A. M. (2011) · Bioengineered Bugs · doi:10.4161/bbug.2.3.15285 · Cited by 107 · see citing works
Genetic Instability in Cyanobacteria – An Elephant in the Room?
Jones, P. R. (2014) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2014.00012 · Cited by 105 · see citing works
Production of Squalene in Synechocystis sp. PCC 6803
Englund, E., Pattanaik, B., Ubhayasekera, S. J. K. A., Stensjö, K., Bergquist, J., & Lindberg, P. (2014) · PLoS ONE · doi:10.1371/journal.pone.0090270 · Cited by 103 · see citing works
Thomas, J., Ughy, B., Lagoutte, B., et al. (2006) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0607718103 · Cited by 97 · see citing works
Miao, R., Liu, X., Englund, E., Lindberg, P., & Lindblad, P. C. (2017) · Metabolic Engineering Communications · doi:10.1016/j.meteno.2017.07.003 · Cited by 87 · see citing works
Erdrich, P., Knoop, H., Steuer, R., & Klamt, S. (2014) · Microbial Cell Factories · doi:10.1186/s12934-014-0128-x · Cited by 85 · see citing works
Regulation of ammonium assimilation in cyanobacteria
Muro‐Pastor, M. I., & Florencio, F. J. (2003) · Plant Physiology and Biochemistry · doi:10.1016/s0981-9428(03)00066-4 · Cited by 84 · see citing works
Computational evaluation of Synechococcus sp. PCC 7002 metabolism for chemical production
Vu, T. T., Hill, E. A., Kucek, L. A., Konopka, A. E., Beliaev, A. S., & Reed, J. L. (2013) · Biotechnology Journal · doi:10.1002/biot.201200315 · Cited by 67 · see citing works
Self-citationClark, R. L., McGinley, L. L., Purdy, H. M., Korosh, T. C., Reed, J. L., Root, T. W., & Pfleger, B. F. (2018) · Metabolic Engineering · doi:10.1016/j.ymben.2018.03.017 · Cited by 66 · see citing works
Self-citationMiao, R., Xie, H., Ho, F. M., & Lindblad, P. C. (2018) · Metabolic Engineering · doi:10.1016/j.ymben.2018.02.014 · Cited by 61 · see citing works
Photosynthetic production of glycerol by a recombinant cyanobacterium
Savakis, P., Tan, X., Du, W., dos Santos, F. B., Lü, X., & Hellingwerf, K. J. (2014) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2014.12.015 · Cited by 58 · see citing works
The future of self-selecting and stable fermentations
Rugbjerg, P., & Olsson, L. (2020) · Journal of Industrial Microbiology & Biotechnology · doi:10.1007/s10295-020-02325-0 · Cited by 43 · see citing works
Engineering Photosynthetic Bioprocesses for Sustainable Chemical Production: A Review
Stephens, S. M., Mahadevan, R., & Allen, D. G. (2021) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2020.610723 · Cited by 39 · see citing works
Synechococcus sp. Strain PCC 7002 nifJ Mutant Lacking Pyruvate:Ferredoxin Oxidoreductase
McNeely, K., Xu, Y., Ananyev, G., Bennette, N. B., Bryant, D. A., & Dismukes, G. C. (2011) · Applied and Environmental Microbiology · doi:10.1128/aem.02792-10 · Cited by 38 · see citing works
Biofuel toxicity and mechanisms of biofuel tolerance in three model cyanobacteria
Ruffing, A. M., & Trahan, C. A. (2014) · Algal Research · doi:10.1016/j.algal.2014.07.006 · Cited by 34 · see citing works
Toward systems metabolic engineering in cyanobacteria
Nogales, J., Guðmundsson, S., & Thiele, I. (2012) · Bioengineered · doi:10.4161/bioe.22792 · Cited by 33 · see citing works
In silico strategies to couple production of bioethanol with growth in cyanobacteria
Testa, R. L., Delpino, C., Estrada, V., & Díaz, M. S. (2019) · Biotechnology and Bioengineering · doi:10.1002/bit.26998 · Cited by 25 · see citing works
Okuhara, H., Matsumura, T., Fujita, Y., et al. (1999) · PLANT PHYSIOLOGY · doi:10.1104/pp.120.1.33 · Cited by 24 · see citing works
Enhancing Photosynthetic Production of Glycogen-Rich Biomass for Use as a Fermentation Feedstock
Comer, A. D., Abraham, J. P., Steiner, A. J., Korosh, T. C., Markley, A. L., & Pfleger, B. F. (2020) · Frontiers in Energy Research · doi:10.3389/fenrg.2020.00093 · Cited by 22 · see citing works
Self-citationSakamoto, T., Inoue‐Sakamoto, K., Persson, S., et al. (2008) · Phycological Research · doi:10.1111/j.1440-1835.2008.00504.x · Cited by 6 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Cyanobacteria as whole-cell factories: Current status and future prospectives
Toepel, J., Karande, R., Klähn, S., & Bühler, B. (2023) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2023.102892 · Cited by 52 · see citing works
Theodosiou, E., Tüllinghoff, A., Toepel, J., & Bühler, B. (2022) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2022.855715 · Cited by 17 · see citing works
Assessing and reducing phenotypic instability in cyanobacteria
Guillaume, M. C., & Branco dos Santos, F. (2023) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2023.102899 · Cited by 11 · see citing works
Carbon capture, storage, and utilization in cyanobacteria
Majhi, B. K. (2026) · Carbon Resources Conversion · doi:10.1016/j.crcon.2025.100388 · Cited by 2 · see citing works
Walshe, J., & Saha, S. K. (2026) · Current Issues in Molecular Biology · doi:10.3390/cimb48060638 · Cited by 1 · see citing works
“Under optimal laboratory conditions, this strain can attain higher biomass productivity than several commonly used cyanobacterial hosts, thereby enhancing its potential for industrial biotechnology applications [64,66].”
- Authors
- Purdy, H. M., Pfleger, B. F., & Reed, J. L.
- Affiliations
- University of Wisconsin System (US)
- University of Wisconsin–Madison (US)
- Corresponding author
- Pfleger, Brian F.
- Venue
- Metabolic Engineering
- Publisher
- Elsevier BV
- Published
- 2022-01
- Volume
- 69
- Pages
- 87-97
- ISSN
- 1096-7176 (ISSN-L)
- PMC
- PMC9026717
- OpenAlex
- W3211861582
- Cited by
- 18
- References
- 54
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 2 · 2023: 7 · 2022: 6
- FWCI
- 1.18
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Metabolic Engineering (acceptedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
- Funders
- US Department of Energy (DE-SC0008103)
- National Institutes of Health (T32 HG002760)
- Biological and Environmental Research
- Office of Science
- College of Engineering, University of Wisconsin-Madison (T32 HG002760)
- 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
- Synechococcus sp. PCC 7002, cyanobacteria, 2-methyl-1-butanol, isobutanol, n-butanol, genome-scale metabolic models
- MeSH terms
- Butanols, NAD, Nitrates, 1-Butanol, Synechococcus
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
