An engineered microbial platform for direct biofuel production from brown macroalgae
Abstract
Prospecting macroalgae (seaweeds) as feedstocks for bioconversion into biofuels and commodity chemical compounds is limited primarily by the availability of tractable microorganisms that can metabolize alginate polysaccharides. Here, we present the discovery of a 36-kilo-base pair DNA fragment from Vibrio splendidus encoding enzymes for alginate transport and metabolism. The genomic integration of this ensemble, together with an engineered system for extracellular alginate depolymerization, generated a microbial platform that can simultaneously degrade, uptake, and metabolize alginate. When further engineered for ethanol synthesis, this platform enables bioethanol production directly from macroalgae via a consolidated process, achieving a titer of 4.7% volume/volume and a yield of 0.281 weight ethanol/weight dry macroalgae (equivalent to ~80% of the maximum theoretical yield from the sugar composition in macroalgae).
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DEHEP is non-enzymatically rearranged into DEHU, which is reduced to KDG by DEHU reductase.
“Subsequently, DEHEP is non-enzymatically rearranged into 4-deoxy-l-erythro-5-hexoseulose uronic acid (DEHU), which is reduced to 2-keto-3-deoxy-gluconate (KDG) by DEHU reductase (EC is not available) (Figure 2F) (Wargacki et al., 2012).”
From The Enzymatic Conversion of Major Algal and Cyanobacterial Carbohydrates to Bioethanol (2016) · cites this paper as Wargacki et al., 2012
A plasmid-based E. coli strain capable of simultaneous degradation and fermentation of alginate to ethanol was engineered using alginate metabolic pathway genes from Vibrio splendidus; it fermented 80% of the maximum theoretical yield and produced 0.281 weight ethanol/weight dry macroalgae.
“Similarly, a plasmid-based E. coli strain, which is capable of simultaneous degradation and fermentation of alginate to ethanol, was engineered using alginate metabolic pathway genes from Vibrio splendidus.This strain, which can metabolize alginate, mannitol, and laminarin, fermented 80% of the maximum theoretical yield and produces 0.281 weight ethanol/weight dry macroalgae (Wargacki et al., 2012”
From The Enzymatic Conversion of Major Algal and Cyanobacterial Carbohydrates to Bioethanol (2016) · cites this paper as Wargacki et al., 2012
Cites
One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products.
Datsenko, K., & Wanner, B. (2000) · Proceedings of the National Academy of Sciences of the United States of America · doi:10.1073/pnas.120163297 · Cited by 15,902 · see citing works
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
Engineering a mevalonate pathway in Escherichia coli for production of terpenoids
Martin, V. J. J., Pitera, D. J., Withers, S. T., Newman, J. D., & Keasling, J. D. (2003) · Nature Biotechnology · doi:10.1038/nbt833 · Cited by 1,720 · see citing works
Isoprenoid Pathway Optimization for Taxol Precursor Overproduction in Escherichia coli
Ajikumar, P. K., Xiao, W., Tyo, K. E. J., Wang, Y., Simeon, F., Leonard, E., Mucha, O., Too, H., Pfeifer, B. A., & Stephanopoulos, G. N. (2010) · Science · doi:10.1126/science.1191652 · Cited by 1,681 · see citing works
Self-citationConsolidated bioprocessing of cellulosic biomass: an update
Lynd, L. R., van Zyl, W. H., McBride, J. E., & Laser, M. S. (2005) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2005.08.009 · Cited by 1,451 · see citing works
Microbial production of fatty-acid-derived fuels and chemicals from plant biomass
Steen, E. J., Kang, Y., Bokinsky, G., Hu, Z., Schirmer, A., McClure, A., del Cardayré, S. B., & Keasling, J. D. (2010) · Nature · doi:10.1038/nature08721 · Cited by 1,338 · see citing works
Feedstocks for Lignocellulosic Biofuels
Somerville, C., Youngs, H., Taylor, C., Davis, S. C., & Long, S. P. (2010) · Science · doi:10.1126/science.1189268 · Cited by 1,307 · see citing works
Metabolic engineering of Escherichia coli for direct production of 1,4-butanediol
Yim, H. H., Haselbeck, R. J., Niu, W., Pujol-Baxley, C., Burgard, A. P., Boldt, J., Khandurina, J., Trawick, J. D., Osterhout, R. E., Stephen, R., et al. (2011) · Nature Chemical Biology · doi:10.1038/nchembio.580 · Cited by 1,119 · see citing works
Microbial Biosynthesis of Alkanes
Schirmer, A., Rude, M. A., Li, X., Popova, E., & del Cardayre, S. B. (2010) · Science · doi:10.1126/science.1187936 · Cited by 1,040 · see citing works
Biosynthesis of Complex Polyketides in a Metabolically Engineered Strain of E. coli
Pfeifer, B. A., Admiraal, S. J., Gramajo, H. C., Cane, D. E., & Khosla, C. (2001) · Science · doi:10.1126/science.1058092 · Cited by 751 · see citing works
Challenges in Engineering Microbes for Biofuels Production
Stephanopoulos, G. N. (2007) · Science · doi:10.1126/science.1139612 · Cited by 731 · see citing works
Wong, T. Y., Preston, L. A., & Schiller, N. L. (2000) · Annual Review of Microbiology · doi:10.1146/annurev.micro.54.1.289 · Cited by 669 · see citing works
Engineered reversal of the β-oxidation cycle for the synthesis of fuels and chemicals
Dellomonaco, C., Clomburg, J. M., Miller, E. N., & González, R. (2011) · Nature · doi:10.1038/nature10333 · Cited by 597 · see citing works
Ohta, K., Beall, D. S., Mejia, J., Shanmugam, K. T., & Ingram, L. O. (1991) · Applied and Environmental Microbiology · doi:10.1128/aem.57.4.893-900.1991 · Cited by 488 · see citing works
Alper, H. S., Miyaoku, K., & Stephanopoulos, G. N. (2005) · Nature Biotechnology · doi:10.1038/nbt1083 · Cited by 433 · see citing works
Engineering for biofuels: exploiting innate microbial capacity or importing biosynthetic potential?
Alper, H. S., & Stephanopoulos, G. N. (2009) · Nature Reviews Microbiology · doi:10.1038/nrmicro2186 · Cited by 384 · see citing works
Adams, J. M. M., Gallagher, J. A., & Donnison, I. S. (2008) · Journal of Applied Phycology · doi:10.1007/s10811-008-9384-7 · Cited by 363 · see citing works
Lau, M. W., & Dale, B. E. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0812364106 · Cited by 341 · see citing works
Enzyme mechanism as a kinetic control element for designing synthetic biofuel pathways
Bond-Watts, B., Bellerose, R. J., & Chang, M. C. Y. (2011) · Nature Chemical Biology · doi:10.1038/nchembio.537 · Cited by 333 · see citing works
Ethanol production from seaweed extract
Horn, S. J., Aasen, I. M., & �stgaard, K. (2000) · Journal of Industrial Microbiology & Biotechnology · doi:10.1038/sj.jim.7000065 · Cited by 310 · see citing works
Macroalgae as a Biomass Feedstock: A Preliminary Analysis
Roesijadi, G., Jones, S. B., Snowden-Swan, L. J., & Zhu, Y. (2010) · doi:10.2172/1006310 · Cited by 269 · see citing works
Strain Improvement of Recombinant Escherichia coli for Efficient Production of Plant Flavonoids
Leonard, E., Yan, Y., Fowler, Z. L., Li, Z., Lim, C.-G., Lim, K. H., & Koffas, M. A. G. (2008) · Molecular Pharmaceutics · doi:10.1021/mp7001472 · Cited by 268 · see citing works
Self-citationBioethanol production from marine biomass alginate by metabolically engineered bacteria
Takeda, H., Yoneyama, F., Kawai, S., Hashimoto, W., & Murata, K. (2011) · Energy & Environmental Science · doi:10.1039/c1ee01236c · Cited by 186 · see citing works
The secretome of the plant pathogenic bacterium Erwinia chrysanthemi
Kazemipour, N., Condemine, G., & Hugouvieux‐Cotte‐Pattat, N. (2004) · PROTEOMICS · doi:10.1002/pmic.200300814 · Cited by 184 · see citing works
Production of ethanol from mannitol by Zymobacter palmae
Horn, S. J., Aasen, I. M., & �stgaard, K. (2000) · Journal of Industrial Microbiology & Biotechnology · doi:10.1038/sj.jim.2900771 · Cited by 179 · see citing works
Klemm, P., Hjerrild, L., Gjermansen, M., & Schembri, M. A. (2003) · Molecular Microbiology · doi:10.1046/j.1365-2958.2003.03833.x · Cited by 179 · see citing works
Henderson, I. R., & Owen, P. (1999) · Journal of Bacteriology · doi:10.1128/jb.181.7.2132-2141.1999 · Cited by 165 · see citing works
Regulation and Function of Ag43 (Flu)
van der Woude, M. W., & Henderson, I. R. (2008) · Annual Review of Microbiology · doi:10.1146/annurev.micro.62.081307.162938 · Cited by 154 · see citing works
He, S. Y., Lindeberg, M., Chatterjee, A., et al. (1991) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.88.3.1079 · Cited by 136 · see citing works
Hashimoto, W., Miyake, O., Momma, K., et al. (2000) · Journal of Bacteriology · doi:10.1128/jb.182.16.4572-4577.2000 · Cited by 130 · see citing works
Yomano, L. P., York, S. W., Shanmugam, K. T., & Ingram, L. O. (2009) · Biotechnology Letters · doi:10.1007/s10529-009-0011-8 · Cited by 123 · see citing works
Zhang, Z., Yu, G., Guan, H., et al. (2004) · Carbohydrate Research · doi:10.1016/j.carres.2004.03.010 · Cited by 111 · see citing works
Purification and Characterization of a Bifunctional Alginate Lyase from Pseudoalteromonas sp. SM0524
Li, J., Dong, S., Song, J., et al. (2011) · Marine Drugs · doi:10.3390/md9010109 · Cited by 106 · see citing works
Construction and Characterization of a 1,3-Propanediol Operon
Skraly, F. A., Lytle, B. L., & Cameron, D. C. (1998) · Applied and Environmental Microbiology · doi:10.1128/aem.64.1.98-105.1998 · Cited by 90 · see citing works
Alginic Acid Metabolism in Bacteria
Preiss, J., & Ashwell, G. (1962) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)93921-9 · Cited by 85 · see citing works
Takase, R., Ochiai, A., Mikami, B., Hashimoto, W., & Murata, K. (2010) · Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics · doi:10.1016/j.bbapap.2010.05.010 · Cited by 78 · see citing works
Antigen 43-Mediated Autotransporter Display, a Versatile Bacterial Cell Surface Presentation System
Kjærgaard, K., Hasman, H., Schembri, M. A., et al. (2002) · Journal of Bacteriology · doi:10.1128/jb.184.15.4197-4204.2002 · Cited by 76 · see citing works
The Oligogalacturonate-specific Porin KdgM of Erwinia chrysanthemi Belongs to a New Porin Family
Blot, N., Berrier, C., Hugouvieux‐Cotte‐Pattat, N., Ghazi, A., & Condemine, G. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m109193200 · Cited by 61 · see citing works
Ochiai, A., Hashimoto, W., & Murata, K. (2006) · Research in Microbiology · doi:10.1016/j.resmic.2006.02.006 · Cited by 54 · see citing works
Hugouvieux‐Cotte‐Pattat, N., & Reverchon, S. (2001) · Molecular Microbiology · doi:10.1046/j.1365-2958.2001.02565.x · Cited by 47 · see citing works
Hashimoto, W., Kawai, S., & Murata, K. (2010) · Bioengineered Bugs · doi:10.4161/bbug.1.2.10322 · Cited by 37 · see citing works
Improved precursor-directed biosynthesis in E. coli via directed evolution
Lee, H., Harvey, C. J. B., Cane, D. E., et al. (2010) · The Journal of Antibiotics · doi:10.1038/ja.2010.129 · Cited by 26 · see citing works
Fujiyama, K., Maki, H., Kinoshita, S., et al. (1995) · FEMS Microbiology Letters · doi:10.1111/j.1574-6968.1995.tb07384.x · Cited by 17 · see citing works
Ochiai, A., Yamasaki, M., Mikami, B., et al. (2006) · Acta Crystallographica Section F Structural Biology and Crystallization Communications · doi:10.1107/s1744309106014333 · Cited by 8 · see citing works
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Chen, C.-Y., Zhao, X.-Q., Yen, H.-W., Ho, S.-H., Cheng, C.-L., Lee, D.-J., Bai, F.-W., & Chang, J.-S. (2013) · Biochemical Engineering Journal · doi:10.1016/j.bej.2013.03.006 · Cited by 724 · see citing works
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Ho, S.-H., Huang, S.-W., Chen, C.-Y., Hasunuma, T., Kondo, A., & Chang, J.-S. (2013) · Bioresource Technology · doi:10.1016/j.biortech.2012.10.015 · Cited by 683 · see citing works
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Jung, K. A., Lim, S.-R., Kim, Y., & Park, J. M. (2013) · Bioresource Technology · doi:10.1016/j.biortech.2012.10.025 · Cited by 487 · see citing works
Kumar, M., Sun, Y., Rathour, R., Pandey, A., Thakur, I. S., & Tsang, D. C. W. (2020) · Science of The Total Environment · doi:10.1016/j.scitotenv.2020.137116 · Cited by 472 · see citing works
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Enquist-Newman, M., Faust, A. M. E., Bravo, D. D., Santos, C. N. S., Raisner, R. M., Hanel, A., Sarvabhowman, P., Le, C., Regitsky, D. D., Cooper, S. R., et al. (2014) · Nature · doi:10.1038/nature12771 · Cited by 351 · see citing works
Self-citationCampbell, I., Macleod, A., Sahlmann, C., Neves, L., Funderud, J., Øverland, M., Hughes, A. D., & Stanley, M. (2019) · Frontiers in Marine Science · doi:10.3389/fmars.2019.00107 · Cited by 328 · see citing works
Macroalgae-derived biofuel: A review of methods of energy extraction from seaweed biomass
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Kumar, S., Gupta, R., Kumar, G., Sahoo, D., & Kuhad, R. C. (2013) · Bioresource Technology · doi:10.1016/j.biortech.2012.10.120 · Cited by 319 · see citing works
Thomas, F., Barbeyron, T., Tonon, T., Génicot, S., Czjzek, M., & Michel, G. (2012) · Environmental Microbiology · doi:10.1111/j.1462-2920.2012.02751.x · Cited by 205 · see citing works
Application of synthetic biology in cyanobacteria and algae
Wang, B., Wang, J., Zhang, W., & Meldrum, D. R. (2012) · Frontiers in Microbiology · doi:10.3389/fmicb.2012.00344 · Cited by 211 · see citing works
Milledge, J. J., Nielsen, B. V., & Bailey, D. (2016) · Reviews in Environmental Science and Bio/Technology · doi:10.1007/s11157-015-9381-7 · Cited by 193 · see citing works
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- Authors
- Wargacki, A. J., Leonard, E., Win, M. N., Regitsky, D. D., Santos, C. N. S., Kim, P. B., Cooper, S. R., Raisner, R. M., Herman, A., Sivitz, A. B., Lakshmanaswamy, A., Kashiyama, Y., Baker, D., & Yoshikuni, Y.
- Affiliations
- Bio Architecture Lab, 604 Bancroft Way, Suite A, Berkeley, CA 94710, USA.
- University of Washington (US)
- Corresponding author
- Wargacki, Adam J.
- Leonard, Effendi
- Win, Maung Nyan
- Yoshikuni, Yasuo
- Venue
- Science
- Published
- 2012-01-20
- Volume
- 335
- Issue
- 6066
- Pages
- 308-313
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W1979652083
- Cited by
- 734
- References
- 44
- Citations by year
- 2026: 27 · 2025: 22 · 2024: 38 · 2023: 34 · 2022: 48 · 2021: 47 · 2020: 49 · 2019: 60 · 2018: 57 · 2017: 49 · 2016: 67 · 2015: 73 · 2014: 64 · 2013: 53 · 2012: 46
- FWCI
- 30.32
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Science (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Advanced Research Projects Agency - Energy (DE-AR0000006)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Physical Sciences › Engineering › Biomedical Engineering › Biofuel production and bioconversion
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Keywords
- brown macroalgae, seaweed biofuels, Vibrio splendidus, alginate depolymerization, ethanol production, microbial engineering, bioethanol yield
- MeSH terms
- Ethanol, Phaeophyceae, Alginates, Bacterial Proteins, Biological Transport, Carrier Proteins, Escherichia coli, Fermentation, Genes, Bacterial, Glucose, Hexuronic Acids, Mannitol, Polysaccharide-Lyases, Seaweed, Vibrio, Open Reading Frames, Biomass, Lactic Acid, Glucuronic Acid, Metabolic Networks and Pathways, Biofuels, Metabolic Engineering
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