Abstract
Photosynthetic chemical production in cyanobacteria is a promising technology for renewable energy, CO2 mitigation, and fossil fuel replacement. Metabolic engineering has enabled a direct biosynthetic process from CO2 fixation to chemical feedstocks and biofuels, without requiring costly production, storage, and breakdown of cellulose or sugars. However, direct production technology is challenged by a need to achieve high-carbon partitioning to products in order to be competitive. This review discusses principles for the design of biosynthetic pathways in cyanobacteria and describes recent advances in relevant genetic tools. This field is at a critical juncture in assessing the strength and feasibility of carbon partitioning. To address this, we have included the stoichiometry of reducing equivalents and carbon conservation for heterologous pathways, and a method for calculating product yields against a theoretical maximum.
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References
Abe K, Sakai Y, Nakashima S, Araki M, Yoshida W, Sode K, Ikebukuro K (2013) Design of riboregulators for control of cyanobacterial (Synechocystis) protein expression. Biotechnol Lett 36:287–294. doi:10.1007/s10529-013-1352-x
Akiyama H, Okuhata H, Onizuka T, Kanai S, Hirano M, Tanaka S, Sasaki K, Miyasaka H (2011) Antibiotics-free stable polyhydroxyalkanoate (PHA) production from carbon dioxide by recombinant cyanobacteria. Bioresour Technol 102(23):11039–11042. doi:10.1016/j.biortech.2011.09.058
Angermayr SA, Paszota M, Hellingwerf KJ (2012) Engineering a cyanobacterial cell factory for production of lactic acid. Appl Environ Microbiol 78(19):7098–7106. doi:10.1128/AEM.01587-12
Atsumi S, Higashide W, Liao JC (2009) Direct photosynthetic recycling of carbon dioxide to isobutyraldehyde. Nat Biotechnol 27(12):1177–1180. doi:10.1038/nbt.1586
Begemann MB, Zess EK, Walters EM, Schmitt EF, Markley AL, Pfleger BF (2013) An organic acid based counter selection system for cyanobacteria. PLoS One 8(10):e76594. doi:10.1371/journal.pone.0076594
Bell-Pedersen D, Cassone VM, Earnest DJ, Golden SS, Hardin PE, Thomas TL, Zoran MJ (2005) Circadian rhythms from multiple oscillators: lessons from diverse organisms. Nat Rev Genet 6(7):544–556
Bentley FK, Melis A (2012) Diffusion-based process for carbon dioxide uptake and isoprene emission in gaseous/aqueous two-phase photobioreactors by photosynthetic microorganisms. Biotechnol Bioeng 109(1):100–109. doi:10.1002/bit.23298
Berla BM, Saha R, Immethun CM, Maranas CD, Moon TS, Pakrasi HB (2013) Synthetic biology of cyanobacteria: unique challenges and opportunities. Front Microbiol 4:246. doi:10.3389/fmicb.2013.00246
Chungjatupornchai W, Fa-aroonsawat S (2013) The rrnA promoter as a tool for the improved expression of heterologous genes in cyanobacteria. Microbiol Res. doi:10.1016/j.micres.2013.09.010
Deng MD, Coleman JR (1999) Ethanol synthesis by genetic engineering in cyanobacteria. Appl Environ Microbiol 65(2):523–528
Dexter J, Fu PC (2009) Metabolic engineering of cyanobacteria for ethanol production. Energy Environ Sci 2(8):857–864. doi:10.1039/B811937f
Ducat DC, Avelar-Rivas JA, Way JC, Silver PA (2012) Rerouting carbon flux to enhance photosynthetic productivity. Appl Environ Microbiol 78(8):2660–2668. doi:10.1128/AEM.07901-11
Field CB (1998) Primary production of the biosphere: integrating terrestrial and oceanic components. Science 281(5374):237–240. doi:10.1126/science.281.5374.237
Fridlyand L, Kaplan A, Reinhold L (1996) Quantitative evaluation of the role of a putative CO2-scavenging entity in the cyanobacterial CO2-concentrating mechanism. Biosystems 37(3):229–238
Gao ZX, Zhao H, Li ZM, Tan XM, Lu XF (2012) Photosynthetic production of ethanol from carbon dioxide in genetically engineered cyanobacteria. Energy Environ Sci 5(12):9857–9865. doi:10.1039/C2ee22675h
Gimpel JA, Specht EA, Georgianna DR, Mayfield SP (2013) Advances in microalgae engineering and synthetic biology applications for biofuel production. Curr Opin Chem Biol 17(3):489–495. doi:10.1016/j.cbpa.2013.03.038
Guerrero F, Carbonell V, Cossu M, Correddu D, Jones PR (2012) Ethylene synthesis and regulated expression of recombinant protein in Synechocystis sp. PCC 6803. PLoS One 7(11):e50470. doi:10.1371/journal.pone.0050470
Hu P, Borglin S, Kamennaya NA, Chen L, Park H, Mahoney L, Kijac A, Shan G, Chavarría KL, Zhang C, Quinn NWT, Wemmer D, Holman H-Y, Jansson C (2013) Metabolic phenotyping of the cyanobacterium Synechocystis 6803 engineered for production of alkanes and free fatty acids. Appl Energy 102:850–859. doi:10.1016/j.apenergy.2012.08.047
Huang HH, Lindblad P (2013) Wide-dynamic-range promoters engineered for cyanobacteria. J Biol Eng 7(1):10. doi:10.1186/1754-1611-7-10
Imamura S, Asayama M (2009) Sigma factors for cyanobacterial transcription. Gene Regul Syst Biol 3:65–87
Iwaki T, Haranoh K, Inoue N, Kojima K, Satoh R, Nishino T, Wada S, Ihara H, Tsuyama S, Kobayashi H, Wadano A (2006) Expression of foreign type I ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39) stimulates photosynthesis in cyanobacterium Synechococcus PCC7942 cells. Photosynth Res 88(3):287–297. doi:10.1007/s11120-006-9048-x
Joseph A, Aikawa S, Sasaki K, Tsuge Y, Matsuda F, Tanaka T, Kondo A (2013) Utilization of lactic acid bacterial genes in Synechocystis sp. PCC 6803 in the production of lactic acid. Biosci Biotechnol Biochem 77(5):966–970. doi:10.1271/bbb.120921
Kaiser BK, Carleton M, Hickman JW, Miller C, Lawson D, Budde M, Warrener P, Paredes A, Mullapudi S, Navarro P, Cross F, Roberts JM (2013) Fatty aldehydes in cyanobacteria are a metabolically flexible precursor for a diversity of biofuel products. PLoS One 8(3):e58307. doi:10.1371/journal.pone.0058307
Kitayama Y, Kondo T, Nakahira Y, Nishimura H, Ohmiya Y, Oyama T (2004) An in vivo dual-reporter system of cyanobacteria using two railroad-worm luciferases with different color emissions. Plant Cell Physiol 45(1):109–113. doi:10.1093/Pcp/Pch001
Klughammer B, Sultemeyer D, Badger MR, Price GD (1999) The involvement of NAD(P)H dehydrogenase subunits, NdhD3 and NdhF3, in high-affinity CO2 uptake in Synechococcus sp. PCC7002 gives evidence for multiple NDH-1 complexes with specific roles in cyanobacteria. Mol Microbiol 32(6):1305–1315
Kopecna J, Komenda J, Bucinska L, Sobotka R (2012) Long-term acclimation of the cyanobacterium Synechocystis sp. PCC 6803 to high light is accompanied by an enhanced production of chlorophyll that is preferentially channeled to trimeric photosystem I. Plant Physiol 160(4):2239–2250. doi:10.1104/pp.112.207274
Kusakabe T, Tatsuke T, Tsuruno K, Hirokawa Y, Atsumi S, Liao JC, Hanai T (2013) Engineering a synthetic pathway in cyanobacteria for isopropanol production directly from carbon dioxide and light. Metab Eng 20C:101–108. doi:10.1016/j.ymben.2013.09.007
Lan EI, Liao JC (2011) Metabolic engineering of cyanobacteria for 1-butanol production from carbon dioxide. Metab Eng 13(4):353–363. doi:10.1016/j.ymben.2011.04.004
Lan EI, Liao JC (2012) ATP drives direct photosynthetic production of 1-butanol in cyanobacteria. Proc Natl Acad Sci USA 109(16):6018–6023. doi:10.1073/pnas.1200074109
Lan EI, Ro SY, Liao JC (2013) Oxygen-tolerant coenzyme A-acylating aldehyde dehydrogenase facilitates efficient photosynthetic n-butanol biosynthesis in cyanobacteria. Energy Environ Sci 6(9):2672. doi:10.1039/c3ee41405a
Lee JW, Na D, Park JM, Lee J, Choi S, Lee SY (2012) Systems metabolic engineering of microorganisms for natural and non-natural chemicals. Nat Chem Biol 8(6):536–546. doi:10.1038/nchembio.970
Li H, Liao JC (2013) Engineering a cyanobacterium as the catalyst for the photosynthetic conversion of CO2 to 1,2-propanediol. Microb Cell Fact 12:4. doi:10.1186/1475-2859-12-4
Lindberg P, Park S, Melis A (2010) Engineering a platform for photosynthetic isoprene production in cyanobacteria, using Synechocystis as the model organism. Metab Eng 12(1):70–79. doi:10.1016/j.ymben.2009.10.001
Liu X, Curtiss R III (2012) Thermorecovery of cyanobacterial fatty acids at elevated temperatures. J Biotechnol 161(4):445–449. doi:10.1016/j.jbiotec.2012.08.013
Liu X, Fallon S, Sheng J, Curtiss R III (2011a) CO2-limitation-inducible Green Recovery of fatty acids from cyanobacterial biomass. Proc Natl Acad Sci USA 108(17):6905–6908. doi:10.1073/pnas.1103016108
Liu X, Sheng J, Curtiss R 3rd (2011b) Fatty acid production in genetically modified cyanobacteria. Proc Natl Acad Sci USA 108(17):6899–6904. doi:10.1073/pnas.1103014108
Liu J, Chen L, Wang J, Qiao J, Zhang W (2012) Proteomic analysis reveals resistance mechanism against biofuel hexane in Synechocystis sp. PCC 6803. Biotechnol Biofuels 5(1):68. doi:10.1186/1754-6834-5-68
Marcus Y, Altman-Gueta H, Wolff Y, Gurevitz M (2011) Rubisco mutagenesis provides new insight into limitations on photosynthesis and growth in Synechocystis PCC6803. J Exp Bot 62(12):4173–4182. doi:10.1093/jxb/err116
Martin W, Rujan T, Richly E, Hansen A, Cornelsen S, Lins T, Leister D, Stoebe B, Hasegawa M, Penny D (2002) Evolutionary analysis of Arabidopsis, cyanobacterial, and chloroplast genomes reveals plastid phylogeny and thousands of cyanobacterial genes in the nucleus. Proc Natl Acad Sci USA 99(19):12246–12251. doi:10.1073/pnas.182432999
Melis A (2013) Carbon partitioning in photosynthesis. Curr Opin Chem Biol 17(3):453–456. doi:10.1016/j.cbpa.2013.03.010
Mendez-Perez D, Begemann MB, Pfleger BF (2011) Modular synthase-encoding gene involved in alpha-olefin biosynthesis in Synechococcus sp. strain PCC 7002. Appl Environ Microbiol 77(12):4264–4267. doi:10.1128/AEM.00467-11
Miyasaka H, Okuhata H, Tanaka S, Onizuka T, Akiyama H (2013) Polyhydroxyalkanoate (PHA) production from carbon dioxide by recombinant cyanobacteria. In: Marian Petre (ed) Environmental biotechnology—new approaches and prospective applications. InTech. doi:10.5772/54705
Nair U, Thomas C, Golden SS (2001) Functional elements of the strong psbAI promoter of Synechococcus elongatus PCC 7942. J Bacteriol 183(5):1740–1747. doi:10.1128/JB.183.5.1740-1747.2001
Nakahira Y, Ogawa A, Asano H, Oyama T, Tozawa Y (2013) Theophylline-dependent riboswitch as a novel genetic tool for strict regulation of protein expression in cyanobacterium Synechococcus elongatus PCC 7942. Plant Cell Physiol 54(10):1724–1735. doi:10.1093/pcp/pct115
Niederholtmeyer H, Wolfstadter BT, Savage DF, Silver PA, Way JC (2010) Engineering cyanobacteria to synthesize and export hydrophilic products. Appl Environ Microbiol 76(11):3462–3466. doi:10.1128/AEM.00202-10
Nozzi NE, Oliver JWK, Atsumi S (2013) Cyanobacteria as a platform for biofuel production. Front Bioeng Biotechnol 1:7. doi:10.3389/fbioe.2013.00007
Oliver JW, Machado IM, Yoneda H, Atsumi S (2013) Cyanobacterial conversion of carbon dioxide to 2,3-butanediol. Proc Natl Acad Sci USA 110(4):1249–1254. doi:10.1073/pnas.1213024110
Oliver JW, Machado IM, Yoneda H, Atsumi S (2014) Combinatorial optimization of cyanobacterial 2,3-butanediol production. Metab Eng 22C:76–82. doi:10.1016/j.ymben.2014.01.001
Ono E, Cuello JL (2007) Carbon dioxide mitigation using thermophilic cyanobacteria. Biosyst Eng 96(1):129–134. doi:10.1016/j.biosystemseng.2006.09.010
Paddock ML, Boyd JS, Adin DM, Golden SS (2013) Active output state of the Synechococcus Kai circadian oscillator. Proc Natl Acad Sci USA 110(40):E3849–E3857. doi:10.1073/pnas.1315170110
Pandelia ME, Li N, Norgaard H, Warui DM, Rajakovich LJ, Chang WC, Booker SJ, Krebs C, Bollinger JM Jr (2013) Substrate-triggered addition of dioxygen to the diferrous cofactor of aldehyde-deformylating oxygenase to form a diferric-peroxide intermediate. J Am Chem Soc 135(42):15801–15812. doi:10.1021/ja405047b
Peralta-Yahya PP, Zhang F, del Cardayre SB, Keasling JD (2012) Microbial engineering for the production of advanced biofuels. Nature 488(7411):320–328. doi:10.1038/nature11478
Qiao J, Wang J, Chen L, Tian X, Huang S, Ren X, Zhang W (2012) Quantitative iTRAQ LC-MS/MS proteomics reveals metabolic responses to biofuel ethanol in cyanobacterial Synechocystis sp. PCC 6803. J Proteome Res 11(11):5286–5300. doi:10.1021/pr300504w
Rabinovitch-Deere CA, Oliver JW, Rodriguez GM, Atsumi S (2013) Synthetic biology and metabolic engineering approaches to produce biofuels. Chem Rev 113(7):4611–4632. doi:10.1021/cr300361t
Reinsvold RE, Jinkerson RE, Radakovits R, Posewitz MC, Basu C (2011) The production of the sesquiterpene beta-caryophyllene in a transgenic strain of the cyanobacterium Synechocystis. J Plant Physiol 168(8):848–852. doi:10.1016/j.jplph.2010.11.006
Rosgaard L, de Porcellinis AJ, Jacobsen JH, Frigaard NU, Sakuragi Y (2012) Bioengineering of carbon fixation, biofuels, and biochemicals in cyanobacteria and plants. J Biotechnol 162(1):134–147. doi:10.1016/j.jbiotec.2012.05.006
Ruffing AM (2013) RNA-Seq analysis and targeted mutagenesis for improved free fatty acid production in an engineered cyanobacterium. Biotechnol Biofuels 6(1):113. doi:10.1186/1754-6834-6-113
Ruffing AM, Jones HD (2012) Physiological effects of free fatty acid production in genetically engineered Synechococcus elongatus PCC 7942. Biotechnol Bioeng 109(9):2190–2199. doi:10.1002/bit.24509
Sakai M, Ogawa T, Matsuoka M, Fukuda H (1997) Photosynthetic conversion of carbon dioxide to ethylene by the recombinant cyanobacterium, Synechococcus sp. PCC 7942, which harbors a gene for the ethylene-forming enzyme of Pseudomonas syringae. J Ferment Bioeng 84(5):434–443. doi:10.1016/S0922-338x(97)82004-1
Savakis PE, Angermayr SA, Hellingwerf KJ (2013) Synthesis of 2,3-butanediol by Synechocystis sp. PCC6803 via heterologous expression of a catabolic pathway from lactic acid- and enterobacteria. Metab Eng 20:121–130. doi:10.1016/j.ymben.2013.09.008
Schirmer A, Rude MA, Li X, Popova E, del Cardayre SB (2010) Microbial biosynthesis of alkanes. Science 329(5991):559–562. doi:10.1126/science.1187936
Shen CR, Liao JC (2012) Photosynthetic production of 2-methyl-1-butanol from CO2 in cyanobacterium Synechococcus elongatus PCC7942 and characterization of the native acetohydroxyacid synthase. Energy Environ Sci 5(11):9574–9583. doi:10.1039/C2ee23148d
Shikanai T, Munekage Y, Kimura K (2002) Regulation of proton-to-electron stoichiometry in photosynthetic electron transport: physiological function in photoprotection. J Plant Res 115(1):0003–0010. doi:10.1007/s102650200001
Takahama K, Matsuoka M, Nagahama K, Ogawa T (2003) Construction and analysis of a recombinant cyanobacterium expressing a chromosomally inserted gene for an ethylene-forming enzyme at the psbAI locus. J Biosci Bioeng 95(3):302–305
Takahashi H, Clowez S, Wollman FA, Vallon O, Rappaport F (2013) Cyclic electron flow is redox-controlled but independent of state transition. Nat Commun 4:1954. doi:10.1038/ncomms2954
Tan X, Yao L, Gao Q, Wang W, Qi F, Lu X (2011) Photosynthesis driven conversion of carbon dioxide to fatty alcohols and hydrocarbons in cyanobacteria. Metab Eng 13(2):169–176. doi:10.1016/j.ymben.2011.01.001
Ungerer J, Tao L, Davis M, Ghirardi M, Maness PC, Yu JP (2012) Sustained photosynthetic conversion of CO2 to ethylene in recombinant cyanobacterium Synechocystis 6803. Energy Environ Sci 5(10):8998–9006. doi:10.1039/C2ee22555g
Varman AM, Xiao Y, Pakrasi HB, Tang YJ (2013) Metabolic engineering of Synechocystis sp. strain PCC 6803 for isobutanol production. Appl Environ Microbiol 79(3):908–914. doi:10.1128/AEM.02827-12
Vermaas W (2013) Solar-powered production of biofuels and other petroleum substitutes by cyanobacteria: stoichiometries of reducing equivalents and chemical energy, and energy conversion efficiency. In: Photosynthesis research for food, fuel and the future. Advanced Topics in Science and Technology in China. Springer, Berlin, p 353–357. doi:10.1007/978-3-642-32034-7_74
Vinyard DJ, Gimpel J, Ananyev GM, Cornejo MA, Golden SS, Mayfield SP, Dismukes GC (2013) Natural variants of photosystem II subunit D1 tune photochemical fitness to solar intensity. J Biol Chem 288(8):5451–5462. doi:10.1074/jbc.M112.394668
Vioque A (2007) Transformation of cyanobacteria. In: León R, Galván A, Fernández E (eds) Transgenic microalgae as green cell factories, vol. 616. Advances in Experimental Medicine and Biology. Springer, New York, p 12–22. doi:10.1007/978-0-387-75532-8_2
Vishnivetskaya T (2009) Viable Cyanobacteria and green algae from the permafrost darkness. In: Margesin R (ed) Permafrost soils, vol. 16. Soil biology. Springer, Berlin, p 73–84. doi:10.1007/978-3-540-69371-0_6
Wang J, Chen L, Huang S, Liu J, Ren X, Tian X, Qiao J, Zhang W (2012) RNA-seq based identification and mutant validation of gene targets related to ethanol resistance in cyanobacterial Synechocystis sp. PCC 6803. Biotechnol Biofuels 5(1):89. doi:10.1186/1754-6834-5-89
Wang B, Pugh S, Nielsen DR, Zhang W, Meldrum DR (2013a) Engineering cyanobacteria for photosynthetic production of 3-hydroxybutyrate directly from CO2. Metab Eng 16:68–77. doi:10.1016/j.ymben.2013.01.001
Wang W, Liu X, Lu X (2013b) Engineering cyanobacteria to improve photosynthetic production of alka(e)nes. Biotechnol Biofuels 6(1):69. doi:10.1186/1754-6834-6-69
Xu Y, Guerra LT, Li Z, Ludwig M, Dismukes GC, Bryant DA (2013) Altered carbohydrate metabolism in glycogen synthase mutants of Synechococcus sp. strain PCC 7002: cell factories for soluble sugars. Metab Eng 16:56–67. doi:10.1016/j.ymben.2012.12.002
Zhou J, Zhang H, Zhang Y, Li Y, Ma Y (2012) Designing and creating a modularized synthetic pathway in cyanobacterium Synechocystis enables production of acetone from carbon dioxide. Metab Eng 14(4):394–400. doi:10.1016/j.ymben.2012.03.005
Zhu H, Ren X, Wang J, Song Z, Shi M, Qiao J, Tian X, Liu J, Chen L, Zhang W (2013) Integrated OMICS guided engineering of biofuel butanol-tolerance in photosynthetic Synechocystis sp. PCC 6803. Biotechnol Biofuels 6(1):106. doi:10.1186/1754-6834-6-106
Acknowledgments
This study was supported by a National Science Foundation grant (1066182). We thank Lisa A. Anderson, Nicole E. Nozzi, and Christine A. Rabinovitch-Deere (UC Davis) for critical reading of the manuscript.
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Oliver, J.W.K., Atsumi, S. Metabolic design for cyanobacterial chemical synthesis. Photosynth Res 120, 249–261 (2014). https://doi.org/10.1007/s11120-014-9997-4
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DOI: https://doi.org/10.1007/s11120-014-9997-4