Advertisement

Planta

, Volume 249, Issue 1, pp 145–154 | Cite as

Engineering cyanobacteria for production of terpenoids

  • Po-Cheng Lin
  • Himadri B. PakrasiEmail author
Review
First Online:
  • 239 Downloads
Part of the following topical collections:
  1. Terpenes and Isoprenoids

Abstract

Main conclusion

This review summarizes recent advances in cyanobacterial terpenoid production. The challenges and opportunities of improving terpenoid production by cyanobacteria are discussed.

Terpenoids are a diverse group of natural products with a variety of commercial applications. With recent advances in synthetic biology and metabolic engineering, microbial terpenoid synthesis is being viewed as a feasible approach for industrial production. Among different microbial hosts, cyanobacteria have the potential of sustainable production of terpenoids using light and CO2. Terpene synthases and the precursor pathways have been expressed in cyanobacteria for enhanced production of various terpene hydrocarbons, including isoprene, limonene, β-phellandrene, and farnesene. However, the productivities need to be further improved for commercial production. Many barriers remain to be overcome in order to efficiently convert CO2 to terpenoids. In this review, we will summarize recent efforts on photosynthetic production of terpenoids and discuss the challenges and opportunities of engineering cyanobacteria for terpenoid bioproduction.

Keywords

Terpenoid Cyanobacteria Metabolic engineering MEP pathway MVA pathway 

Abbreviations

DMAPP

Dimethylallyl pyrophosphate

DXS

1-Deoxy-d-xylulose 5-phosphate synthase

GAP

Glyceraldehyde 3-phosphate

IDI

Isopentenyl diphosphate isomerase

IPP

Isopentenyl pyrophosphate

IspS

Isoprene synthase

MEP

2-C-Methyl-d-erythritol-4-phosphate

MVA

Mevalonate

This is a preview of subscription content, log in to check access.

Notes

Acknowledgements

Funding to support this work was provided by the Office of Science (BER), U. S. Department of Energy, to HBP. PCL was supported by a fellowship from the McDonnell International Scholars Academy at Washington University.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Abernathy MH, Yu J, Ma F, Liberton M, Ungerer J, Hollinshead WD, Gopalakrishnan S, He L, Maranas CD, Pakrasi HB, Allen DK, Tang YJ (2017) Deciphering cyanobacterial phenotypes for fast photoautotrophic growth via isotopically nonstationary metabolic flux analysis. Biotechnol Biofuels 10:273.  https://doi.org/10.1186/s13068-017-0958-y Google Scholar
  2. Ajikumar PK, Tyo K, Carlsen S, Mucha O, Phon TH, Stephanopoulos G (2008) Terpenoids: opportunities for biosynthesis of natural product drugs using engineered microorganisms. Mol Pharm 5(2):167–190Google Scholar
  3. Ajikumar PK, Xiao WH, Tyo KE, Wang Y, Simeon F, Leonard E, Mucha O, Phon TH, Pfeifer B, Stephanopoulos G (2010) Isoprenoid pathway optimization for Taxol precursor overproduction in Escherichia coli. Science 330(6000):70–74.  https://doi.org/10.1126/science.1191652 Google Scholar
  4. Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold CJ, Lee TS (2013) Metabolic engineering of Escherichia coli for limonene and perillyl alcohol production. Metab Eng 19:33–41.  https://doi.org/10.1016/j.ymben.2013.05.004 Google Scholar
  5. Banerjee A, Sharkey T (2014) Methylerythritol 4-phosphate (MEP) pathway metabolic regulation. Nat Prod Rep 31(8):1043–1055Google Scholar
  6. Banerjee A, Wu Y, Banerjee R, Li Y, Yan H, Sharkey TD (2013) Feedback inhibition of deoxy-d-xylulose-5-phosphate synthase regulates the methylerythritol 4-phosphate pathway. J Biol Chem 288(23):16926–16936.  https://doi.org/10.1074/jbc.M113.464636 Google Scholar
  7. 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):100–109Google Scholar
  8. Bentley FK, Zurbriggen A, Melis A (2014) Heterologous expression of the mevalonic acid pathway in cyanobacteria enhances endogenous carbon partitioning to isoprene. Mol Plant 7(1):71–86.  https://doi.org/10.1093/mp/sst134 Google Scholar
  9. Betterle N, Melis A (2018) Heterologous leader sequences in fusion constructs enhance expression of geranyl diphosphate synthase and yield of β-phellandrene production in cyanobacteria (Synechocystis). ACS Synth Biol 7(3):912–921Google Scholar
  10. Breitmaier E (2006) Terpenes: importance, general structure, and biosynthesis. In: Terpenes, chap 1. Wiley, WeinheimGoogle Scholar
  11. Brenner MP, Bildsten L, Dyson F, Fortson N, Garwin R, Grober R, Hemley R, Hwa T, Joyce G, Katz J (2006) Engineering microorganisms for energy production. DTIC DocumentGoogle Scholar
  12. Carrieri D, Paddock T, Maness P-C, Seibert M, Yu J (2012) Photo-catalytic conversion of carbon dioxide to organic acids by a recombinant cyanobacterium incapable of glycogen storage. Energy Environ Sci 5(11):9457–9461Google Scholar
  13. Chaves JE, Rueda-Romero P, Kirst H, Melis A (2017) Engineering isoprene synthase expression and activity in cyanobacteria. ACS Synth Biol 6(12):2281–2292.  https://doi.org/10.1021/acssynbio.7b00214 Google Scholar
  14. Choi SY, Lee HJ, Choi J, Kim J, Sim SJ, Um Y, Kim Y, Lee TS, Keasling JD, Woo HM (2016) Photosynthetic conversion of CO 2 to farnesyl diphosphate-derived phytochemicals (amorpha-4, 11-diene and squalene) by engineered cyanobacteria. Biotechnol Biofuels 9(1):202Google Scholar
  15. Chuck CJ, Donnelly J (2014) The compatibility of potential bioderived fuels with Jet A-1 aviation kerosene. Appl Energy 118:83–91Google Scholar
  16. Ciddi V, Srinivasan V, Shuler M (1995) Elicitation of Taxus sp. cell cultures for production of taxol. Biotechnol Lett 17(12):1343–1346Google Scholar
  17. Colby SM, Alonso WR, Katahira EJ, McGarvey DJ, Croteau R (1993) 4S-limonene synthase from the oil glands of spearmint (Mentha spicata). cDNA isolation, characterization, and bacterial expression of the catalytically active monoterpene cyclase. J Biol Chem 268(31):23016–23024Google Scholar
  18. Davies FK, Work VH, Beliaev AS, Posewitz MC (2014) Engineering limonene and bisabolene production in wild type and a glycogen-deficient mutant of Synechococcus sp. PCC 7002. Front Bioeng Biotechnol 2:21.  https://doi.org/10.3389/fbioe.2014.00021 Google Scholar
  19. Dempo Y, Ohta E, Nakayama Y, Bamba T, Fukusaki E (2014) Molar-based targeted metabolic profiling of cyanobacterial strains with potential for biological production. Metabolites 4(2):499–516.  https://doi.org/10.3390/metabo4020499 Google Scholar
  20. Dexter J, Fu P (2009) Metabolic engineering of cyanobacteria for ethanol production. Energy Environ Sci 2(8):857–864Google Scholar
  21. Dismukes GC, Carrieri D, Bennette N, Ananyev GM, Posewitz MC (2008) Aquatic phototrophs: efficient alternatives to land-based crops for biofuels. Curr Opin Biotechnol 19(3):235–240.  https://doi.org/10.1016/j.copbio.2008.05.007 Google Scholar
  22. Englund E, Andersen-Ranberg J, Miao R, Hamberger B, Lindberg P (2015) Metabolic engineering of Synechocystis sp. PCC 6803 for production of the plant diterpenoid manoyl oxide. ACS Synth Biol 4(12):1270–1278.  https://doi.org/10.1021/acssynbio.5b00070 Google Scholar
  23. Englund E, Shabestary K, Hudson EP, Lindberg P (2018) Systematic overexpression study to find target enzymes enhancing production of terpenes in Synechocystis PCC 6803, using isoprene as a model compound. Metab Eng 49:164–177.  https://doi.org/10.1016/j.ymben.2018.07.004 Google Scholar
  24. Ershov YV, Gantt RR, Cunningham FX Jr, Gantt E (2002) Isoprenoid biosynthesis in Synechocystis sp. strain PCC6803 is stimulated by compounds of the pentose phosphate cycle but not by pyruvate or deoxyxylulose-5-phosphate. J Bacteriol 184(18):5045–5051Google Scholar
  25. Formighieri C, Melis A (2015) A phycocyanin phellandrene synthase fusion enhances recombinant protein expression and β-phellandrene (monoterpene) hydrocarbons production in Synechocystis (cyanobacteria). Metab Eng 32:116–124Google Scholar
  26. Formighieri C, Melis A (2016) Sustainable heterologous production of terpene hydrocarbons in cyanobacteria. Photosynth Res 130(1–3):123–135.  https://doi.org/10.1007/s11120-016-0233-2 Google Scholar
  27. Gao X, Gao F, Liu D, Zhang H, Nie X, Yang C (2016) Engineering the methylerythritol phosphate pathway in cyanobacteria for photosynthetic isoprene production from CO2. Energy Environ Sci 9(4):1400–1411Google Scholar
  28. Gordon GC, Korosh TC, Cameron JC, Markley AL, Begemann MB, Pfleger BF (2016) CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng 38:170–179Google Scholar
  29. Halfmann C, Gu L, Gibbons W, Zhou R (2014a) Genetically engineering cyanobacteria to convert CO2, water, and light into the long-chain hydrocarbon farnesene. Appl Microbiol Biotechnol 98(23):9869–9877Google Scholar
  30. Halfmann C, Gu L, Zhou R (2014b) Engineering cyanobacteria for the production of a cyclic hydrocarbon fuel from CO2 and H2O. Green Chem 16(6):3175–3185Google Scholar
  31. Hickman JW, Kotovic KM, Miller C, Warrener P, Kaiser B, Jurista T, Budde M, Cross F, Roberts JM, Carleton M (2013) Glycogen synthesis is a required component of the nitrogen stress response in Synechococcus elongatus PCC 7942. Algal Res 2(2):98–106Google Scholar
  32. Huang HH, Lindblad P (2013) Wide-dynamic-range promoters engineered for cyanobacteria. J Biol Eng 7(1):10.  https://doi.org/10.1186/1754-1611-7-10 Google Scholar
  33. Kanno M, Carroll AL, Atsumi S (2017) Global metabolic rewiring for improved CO2 fixation and chemical production in cyanobacteria. Nat Commun 8:14724.  https://doi.org/10.1038/ncomms14724 Google Scholar
  34. Kim SW, Keasling JD (2001) Metabolic engineering of the nonmevalonate isopentenyl diphosphate synthesis pathway in Escherichia coli enhances lycopene production. Biotechnol Bioeng 72(4):408–415Google Scholar
  35. Kirby J, Nishimoto M, Chow RW, Baidoo EE, Wang G, Martin J, Schackwitz W, Chan R, Fortman JL, Keasling JD (2015) Enhancing terpene yield from sugars via novel routes to 1-deoxy-d-xylulose 5-phosphate. Appl Environ Microbiol 81(1):130–138.  https://doi.org/10.1128/AEM.02920-14 Google Scholar
  36. Kiyota H, Okuda Y, Ito M, Hirai MY, Ikeuchi M (2014) Engineering of cyanobacteria for the photosynthetic production of limonene from CO2. J Biotechnol 185:1–7.  https://doi.org/10.1016/j.jbiotec.2014.05.025 Google Scholar
  37. Knoot CJ, Ungerer J, Wangikar PP, Pakrasi HB (2018) Cyanobacteria: promising biocatalysts for sustainable chemical production. J Biol Chem 293(14):5044–5052Google Scholar
  38. Li X, Shen CR, Liao JC (2014) Isobutanol production as an alternative metabolic sink to rescue the growth deficiency of the glycogen mutant of Synechococcus elongatus PCC 7942. Photosynth Res 120(3):301–310.  https://doi.org/10.1007/s11120-014-9987-6 Google Scholar
  39. Li H, Shen CR, Huang C-H, Sung L-Y, Wu M-Y, Hu Y-C (2016) CRISPR-Cas9 for the genome engineering of cyanobacteria and succinate production. Metab Eng 38:293–302Google Scholar
  40. Li S, Sun T, Xu C, Chen L, Zhang W (2018) Development and optimization of genetic toolboxes for a fast-growing cyanobacterium Synechococcus elongatus UTEX 2973. Metab Eng 48:163–174Google Scholar
  41. Lin P-C, Saha R, Zhang F, Pakrasi HB (2017) Metabolic engineering of the pentose phosphate pathway for enhanced limonene production in the cyanobacterium Synechocystis sp. PCC 6803. Sci Rep 7(1):17503.  https://doi.org/10.1038/s41598-017-17831-y Google Scholar
  42. Liu D, Pakrasi HB (2018) Exploring native genetic elements as plug-in tools for synthetic biology in the cyanobacterium Synechocystis sp PCC 6803. Microb Cell Fact 17(1):48Google Scholar
  43. Liu X, Sheng J, Curtiss R III (2011) Fatty acid production in genetically modified cyanobacteria. Proc Natl Acad Sci 108:6899–6904Google Scholar
  44. Lucker J, El Tamer MK, Schwab W, Verstappen FW, van der Plas LH, Bouwmeester HJ, Verhoeven HA (2002) Monoterpene biosynthesis in lemon (Citrus limon). cDNA isolation and functional analysis of four monoterpene synthases. Eur J Biochem FEBS 269(13):3160–3171Google Scholar
  45. Markley AL, Begemann MB, Clarke RE, Gordon GC, Pfleger BF (2015) Synthetic biology toolbox for controlling gene expression in the cyanobacterium Synechococcus sp. strain PCC 7002. ACS Synth Biol 4(5):595–603.  https://doi.org/10.1021/sb500260k Google Scholar
  46. Martin VJ, Pitera DJ, Withers ST, Newman JD, Keasling JD (2003) Engineering a mevalonate pathway in Escherichia coli for production of terpenoids. Nat Biotechnol 21(7):796–802.  https://doi.org/10.1038/nbt833 Google Scholar
  47. Melis A (2013) Carbon partitioning in photosynthesis. Curr Opin Chem Biol 17(3):453–456.  https://doi.org/10.1016/j.cbpa.2013.03.010 Google Scholar
  48. Niu FX, Lu Q, Bu YF, Liu JZ (2017) Metabolic engineering for the microbial production of isoprenoids: carotenoids and isoprenoid-based biofuels. Synth Syst Biotechnol 2(3):167–175.  https://doi.org/10.1016/j.synbio.2017.08.001 Google Scholar
  49. 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.  https://doi.org/10.1073/pnas.1213024110 Google Scholar
  50. Oliver JW, Machado IM, Yoneda H, Atsumi S (2014) Combinatorial optimization of cyanobacterial 2,3-butanediol production. Metab Eng 22:76–82.  https://doi.org/10.1016/j.ymben.2014.01.001 Google Scholar
  51. Paddon CJ, Keasling JD (2014) Semi-synthetic artemisinin: a model for the use of synthetic biology in pharmaceutical development. Nat Rev Microbiol 12(5):355Google Scholar
  52. Sengupta A, Pakrasi HB, Wangikar PP (2018) Recent advances in synthetic biology of cyanobacteria. Appl Microbiol Biotechnol.  https://doi.org/10.1007/s00253-018-9046-x Google Scholar
  53. Sharkey TD, Wiberley AE, Donohue AR (2008) Isoprene emission from plants: why and how. Ann Bot 101(1):5–18.  https://doi.org/10.1093/aob/mcm240 Google Scholar
  54. Siwko ME, Marrink SJ, de Vries AH, Kozubek A, Uiterkamp AJS, Mark AE (2007) Does isoprene protect plant membranes from thermal shock? A molecular dynamics study. Biochim Biophys Acta (BBA) Biomembr 1768(2):198–206Google Scholar
  55. Song K, Tan X, Liang Y, Lu X (2016) The potential of Synechococcus elongatus UTEX 2973 for sugar feedstock production. Appl Microbiol Biotechnol 100(18):7865–7875.  https://doi.org/10.1007/s00253-016-7510-z Google Scholar
  56. Toth TN, Chukhutsina V, Domonkos I, Knoppova J, Komenda J, Kis M, Lenart Z, Garab G, Kovacs L, Gombos Z, van Amerongen H (2015) Carotenoids are essential for the assembly of cyanobacterial photosynthetic complexes. Biochem Biophys Acta 1847(10):1153–1165.  https://doi.org/10.1016/j.bbabio.2015.05.020 Google Scholar
  57. Tracy NI, Chen D, Crunkleton DW, Price GL (2009) Hydrogenated monoterpenes as diesel fuel additives. Fuel 88(11):2238–2240Google Scholar
  58. Ungerer J, Pakrasi HB (2016) Cpf1 is a versatile tool for CRISPR genome editing across diverse species of cyanobacteria. Sci Rep 6:39681Google Scholar
  59. Ungerer J, Tao L, Davis M, Ghirardi M, Maness P-C, Yu J (2012) Sustained photosynthetic conversion of CO2 to ethylene in recombinant cyanobacterium Synechocystis 6803. Energy Environ Sci 5(10):8998–9006Google Scholar
  60. Ungerer J, Lin PC, Chen HY, Pakrasi HB (2018) Adjustments to photosystem stoichiometry and electron transfer proteins are key to the remarkably fast growth of the cyanobacterium Synechococcus elongatus UTEX 2973. mBio 9(1):e02327-17.  https://doi.org/10.1128/mbio.02327-17 Google Scholar
  61. van der Woude AD, Angermayr SA, Puthan Veetil V, Osnato A, Hellingwerf KJ (2014) Carbon sink removal: increased photosynthetic production of lactic acid by Synechocystis sp. PCC6803 in a glycogen storage mutant. J Biotechnol 184:100–102.  https://doi.org/10.1016/j.jbiotec.2014.04.029 Google Scholar
  62. Wang X, Liu W, Xin C, Zheng Y, Cheng Y, Sun S, Li R, Zhu XG, Dai SY, Rentzepis PM, Yuan JS (2016) Enhanced limonene production in cyanobacteria reveals photosynthesis limitations. Proc Natl Acad Sci USA 113(50):14225–14230.  https://doi.org/10.1073/pnas.1613340113 Google Scholar
  63. Wang B, Eckert C, Maness PC, Yu J (2018) A genetic toolbox for modulating the expression of heterologous genes in the cyanobacterium Synechocystis sp. PCC 6803. ACS Synth Biol 7(1):276–286.  https://doi.org/10.1021/acssynbio.7b00297 Google Scholar
  64. Wendt KE, Ungerer J, Cobb RE, Zhao H, Pakrasi HB (2016) CRISPR/Cas9 mediated targeted mutagenesis of the fast growing cyanobacterium Synechococcus elongatus UTEX 2973. Microb Cell Fact 15(1):115Google Scholar
  65. Yoo SH, Keppel C, Spalding M, Jane JL (2007) Effects of growth condition on the structure of glycogen produced in cyanobacterium Synechocystis sp. PCC6803. Int J Biol Macromol 40(5):498–504.  https://doi.org/10.1016/j.ijbiomac.2006.11.009 Google Scholar
  66. Young JD, Shastri AA, Stephanopoulos G, Morgan JA (2011) Mapping photoautotrophic metabolism with isotopically nonstationary 13C flux analysis. Metab Eng 13(6):656–665.  https://doi.org/10.1016/j.ymben.2011.08.002 Google Scholar
  67. Yu J, Liberton M, Cliften PF, Head RD, Jacobs JM, Smith RD, Koppenaal DW, Brand JJ, Pakrasi HB (2015) Synechococcus elongatus UTEX 2973, a fast growing cyanobacterial chassis for biosynthesis using light and CO(2). Sci Rep 5:8132.  https://doi.org/10.1038/srep08132 Google Scholar
  68. Zhou C, Li Z, Wiberley-Bradford AE, Weise SE, Sharkey TD (2013) Isopentenyl diphosphate and dimethylallyl diphosphate/isopentenyl diphosphate ratio measured with recombinant isopentenyl diphosphate isomerase and isoprene synthase. Anal Biochem 440(2):130–136.  https://doi.org/10.1016/j.ab.2013.05.028 Google Scholar
  69. Zhou J, Wang C, Yang L, Choi E-S, Kim S-W (2015) Geranyl diphosphate synthase: an important regulation point in balancing a recombinant monoterpene pathway in Escherichia coli. Enzyme Microb Technol 68:50–55Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Energy, Environmental and Chemical EngineeringWashington UniversitySt. LouisUSA
  2. 2.Department of BiologyWashington UniversitySt. LouisUSA

Personalised recommendations

Cite article

Buy options