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plant as bioreactor

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  • 1. Plant as Bioreactor
    Submitted by : Arun & Udhaya
    1
  • 2. Introduction
    • A device in which a substrate of low value is utilized by living cells to generate products of higher value.
    • 3. From earlier days - Microbes &animal cell culture used to produce biomolecules.
    • 4. Advancement in Plant genetic engineering : Possible to use as Bioreactors.
    • 5. Plants are important food resources from the earliest times.
    • 6. Plants as bioreactors: Biomolecules, industrial, chemical & pharmaceutical etc.
    • 7. Number of products are successfully produced in plants using this ‘molecular farming’
    2
  • 8. Comparison with other production system
    • Fast biomass build up
    • 9. Post translational modifications
    • 10. Easy storage and distribution
    • 11. Low upstream production cost
    • 12. Transgenic animals as production systems: Public and ethical concerns
    Types of Plant bioreactors
    • Seed-based plant bioreactors
    • 13. Plant Suspension Cultures
    • 14. Hairy Root System Bioreactor
    • 15. Chloroplast bioreactor
    3
  • 16. Advantages : Plant as Bioreactor
    • Low cost source.
    • 17. Simple & Cost effective.
    • 18. Plant pathogens do not infect humans or animals.
    • 19. Easy scale up & rapid harvesting.
    • 20. Chimeric plant viruses can be used in production of vaccines.
    • 21. Produce large biomass.
    • 22. Easy storage for long time.
    4
  • 23. Key process…
    5
  • 24. Production platforms
    • Cereals:Maize, Rice, Barley and Wheat are used.
    • 25. High seed protein content (7 to 10%).
    • 26. High biomass yield.
    • 27. Ease of transformation.
    • 28. Ease of scale-up.
    • 29. Major disadvantageof Maize is: Cross pollination
    • 30. But ricehas advantage of self-pollination.
    • 31. Rice as bioreactors : Oral delivery system for vaccine antigens, immunotherapy and therapeutic proteins are recent advances.
    6
    Streatfield SJ et al., 2003
  • 32. 7
    Production platform Cont…
    Ventria Bioscience (US Based Company) : Two rice-derived proteins
    • Human lactoferrin and lysozyme
    • 33. Have received regulatory approvals, and lactoferrin has already been marketed in bulk for bioprocessing.
  • Production platforms (Cont.…)
    Legumes:
    • Soybean and pea: Seeds have high protein content (20–40%).
    • 34. Both self-pollinating plants & low risk of contamination.
    • 35. Soybean : Express a humanized antibody against Herpes simplex virus. Bovine casein, and a Human growth hormone. (Russell et al., 2005)
    Oil seeds:
    • Safflower and rapeseed : Rich in seed oil & inexpensive downstream.
    • 36. Safflower : High protein yield, Low acreage, and Self-pollinating.
    • 37. Oilseed-derived protein: Commercial production of Hirudin, an anti-coagulant for treatment of thrombosis in rapeseed by SemBioSys(Bootheet al., 1997)
    8
  • 38. Where they are produced?
    Shown are various Intracellular organelles or Extracellular spaces (ES) that can be used to store the recombinant proteins expressed in a plant bioreactor.
    Targeting strategies in plant bioreactors.
    G- Golgi; PSV- Protein storage vacuole;
    OB - Oil body; C- Chloroplast;
    ES- Extracellular space;
    PVC - Prevacuolar compartment.
    9
  • 39. Products obtained…
    Plants genetically engineered to make products that are not of plant origin
    Products:
    • Vaccines antigens
    • 40. Therapeutics products
    • 41. Nutritional components
    • 42. Industrial products
    • 43. Bio plastics
    10
    Tree depicting biotechnological advances using plants as bioreactors
    Source: www.plantbioreactor.co.in/images/00_112.jpg
  • 44. Products : Plant as bioreactor
    Vaccine antigens:
    • It is an antigenic preparation : Immunity against a disease.
    • 45. Recent Development : “Edible Vaccines” now more popular as “Plant Vaccines”
    • 46. From Seeds, freeze-dried fruits and leaves: powder form vaccines are produced.
    • 47. Antigenic determinantsfor Pathogens causing diseases have been produced from plants.(Diarrhea, anthrax, rabies, cancer, HIV, tuberculosis etc)
    • 48. Antigens like Insulin, rotavirus enterotoxin, anthrax lethal factor, HIV antigen, foot and mouth disease virus antigen, heat stable toxin have been produced in plants as a fusion partner of CTB or LTB.
    11
    http://www.dowagro.com/uk/media/General/20061017.htm, Khandelwalet al., 2003; Sharma et al., 2004, Streatfield and Howard, 2003, Tiwariet al., 2009 and Youm et al., 2008.
  • 49. Products : Plant as bioreactor(Cont.…)
    Therapeutic products:
    • Diagnostic proteins (antibodies and enzymes), replacement proteins, immune system stimulator/suppressants, biopolymers and adhesive proteins are produced in plants.
    • 50. Production of immunoglobulin fragments and their assembly in plants –reported in tobacco.
    • 51. Recently known as “plantibodies” – immunochromatography or medical therapy.
    12
    Schillberget al., 1999, Ma et al., 2003, Goldstein and Thomas, 2004
  • 52. Products : Plant as bioreactor
    Therapeutic products:
    • Many form of recombinants antibodies are produced in plants.
    • 53. (Full size recombinant antibody, chimeric antibody, secretory antibody)
    • 54. Sub cellular destination - high level expression.
    • 55. Further research is concentrated on Humanizing the Plant N-glycans.
    13
    Schillberget al., 1999, Ma et al., 2003, Goldstein and Thomas, 2004
  • 56. Products : Plant as bioreactor(Cont.…)
    Nutritional components:
    • It can provide most of the nutrients required in the human diet.
    • 57. Plant have been engineered to increase accumulation of:
    β-carotene(Naqvi et al., 2009), Lycopene(Fraser et al., 2002), Flavonoid(Butelli et al., 2008), Nutraceuticals (Kang et al., 2009), Fatty acid (Hoffmann et al., 2008), Vitamins (Nunes et al., 2009), Minerals(Lee & An, 2009) & Carbohydrates (Regina et al., 2006).
    Biodegradable plastics:
    • Polyhydroxyalkanoates: biodegradable polymers which occur naturally in plants.
    Plant was engineered to produce PHAs or PHBs in the various plant cell compartments.
    • When PHB expression targeted to cytoplasm, accumulation level was low.
    • 58. Expression was increased by targeting plastids, (40% of dry weight was obtained).
    14
  • 59. Products : Plant as bioreactor(Cont.…)
    Industrial products:
    • First produced protein : Human growth hormones in tobacco.
    • 60. Most expensive Drug – hGC (Human glucocerebrosidase) in tobacco (Kaiser 2008).
    • 61. hST (Human somatotropin) was produced in tobacco (Staubet al. 2000).
    • 62. Which treats : Turner syndrome, Chronical renal failure & Dwarfism in Children.
    • 63. rHLF (Recombinant human lactoferrin) : Produced from dehusked rice grain(Nandi et al., 2002). Which is identical to native HLF.
    • 64. Synthetic fiber: Produced from Potato and tobacco. (Schelleret al., 2001)
    15
  • 65. Conclusion…
    • More and more uses of plant bioreactors are coming up these days.
    • 66. Plant bioreactors : Investigated for making enzymes (Suitable for Food).
    • 67. Another use of plants is to make genetically engineered plants that can produce seeds which can function as a delivery mechanism for various industrial enzymes.
    • 68. As you can see these processes go far beyond the application of biotechnology in traditional agriculture, and so today, transgenic plants can produce on a mass scale proteins for agricultural, veterinary and pharmaceutical use.
    16
  • 69. Conclusion (Cont.…)
    17
    Table source: Yansongmiaoet al., 2008
  • 70. Problems need to be addressed
    • Storage issues related to transgenic fruits or leaves.
    • 71. Most inserted genes are expressed at very low level in plants.
    • 72. Enhancing the stability of products obtained.
    • 73. Standardization of dosage in case of edible vaccine.
    • 74. Examining issues related to commercialization.
    • 75. Issues relating to the ethical, social, biosafety and environmental impact.
    • 76. Some plants produce allergenic compounds.
    18
  • 77. References
    Boothe JG, Parmenter DL, Saponja JA. (1997) Molecular farming in plants: oilseeds as vehicles for the production of pharmaceutical proteins. Drug Dev Res, 42:172–81.
    Butelli E, Titta L, Giorgio M, Mock HP, Matros A, Peterek S, (2008). Enrichment of tomato fruit with health-promoting anthocyanins by expression of select transcription factors. Nat Biotechnol, 26:1301–8.
    Dow AgroSciences. Dow AgroSciences achieves world's first registration for plant-made vaccines. 2006 Press release (http://www.dowagro.com/animalhealth/resources/news/20060131b.htm).
    Fraser PD, Romer S, Shipton CA, Mills PB, Kiano JW, Misawa N (2009). Evaluation of transgenic tomato plants expressing an additional phytoene synthase in a fruit specific manner. ProcNatlAcadSci USA , 99: 1092–7.
    Goldstein DA, Thomas JA. (2004), Biopharmaceuticals derived from genetically modified plants. QJM, 97: 705–16.
    Hiatt A, Cafferkey R, Bowdish K. (1989), Production of antibodies in transgenic plants. Nature, 342: 76–8.
    Hoffmann M, Wagner M, Abbadi A, Fulda M, Feussner I. (2008), Metabolic engineering of omega3-very long chain polyunsaturated fatty acid production by an exclusively acyl-CoA-dependent pathway. J BiolChem, 283: 22352–62.
    Hood EE, Witcher DR, Maddock S, Meyer T, Baszczynski C, Bailey M, (1997), Commercial production of avidin from transgenic maize: characterization of transformant, production, processing, extraction and purification. Mol Breed, 3: 291–306.
    19
  • 78. References(Cont.…)
    Kang K, Lee K, Sohna S, Parka S, Lee S, Kima S, (2009), Ectopic expression of serotonin hydroxycinnamoyltransferase and differential production of phenylpropanoid amides in transgenic tomato tissues. SciHortic, 120: 504–10.
    Khandelwal A, Sita GL, Shaila MS. (2003), Expression of hemagglutinin protein of rinderpest virus in transgenic tobacco and immunogenicity of plant-derived protein in a mouse model. Virology, 308: 207–15.
    Lee S, An G. (2009), Over-expression of OsIRT1 leads to increased iron and zinc accumulations in rice. Plant Cell Environ, 32: 408–16.
    Ma JK, Drake PM, Christou P. (2003), the production of recombinant pharmaceutical proteins in plants. Nat Rev Genet, 4: 794–805.
    Naqvi S, Zhu C, Farre G, Ramessar K, Bassie L, Breitenbach J, (2009), Transgenic multivitamin corn through biofortification of endosperm with three vitamins representing three distinct metabolic pathways. ProcNatlAcadSci USA, 106: 7762–7.
    Nunes AC, Kalkmann DC, Aragão FJ.(2009), Folatebiofortification of lettuce by expression of a codon optimized chicken GTP cyclohydrolase I gene. Transgenic Res, doi:10.1007/s11248-009-9256-1.
    Pujol M, Ramírez NI, Ayala M, Gavilondo JV, Valdés R, Rodríguez M, (2005), An integral approach towards a practical application for a plant-made monoclonal antibody in vaccine purification. Vaccine, 23: 1833–7.
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  • 79. References (Cont.…)
    Regina A, Bird A, Topping D, Bowden S, Freeman J, Barsby T, (2006). High-amylose wheat generated by RNA interference improves indices of large-bowel health in rats. Proc. Natl. Acad. Sci. USA, 103: 3546–51.
    Russell DA, Spatola LA, Dian T, Paradkar VM, Dufield DR, Carrol JA, (2005). Host limits to accurate human growth hormone production in multiple plant systems. BiotechnolBioeng, 89: 775–82.
    Schillberg S, Zimmermann S, Voss A, Fischer R. (1999), Apoplastic and cytosolic expression of full-size antibodies and antibody fragments in Nicotianatabacum. Transgenic Res. 8:255–63.
    Sharma AK, Jani D, Tyagi AK. (2004), Transgenic plants as bioreactors. Ind J Biotechnol. 3: 274–90.
    Streatfield SJ, Howard JA. (2003), Plant-based vaccines. Int J Parasitol, 33: 479–93.
    Tiwari S, Verma PC, Singh PK, Tuli R. (2009), Plants as bioreactors for the production of vaccine antigens. Biotech adv, 449–67.
    Woodard SL, Mayor JM, Bailey MR, Barker DK, Love RT, Lane JR, (2003), Maize (Zea mays)- derived bovine trypsin: characterization of the first large-scale, commercial protein product from transgenic plants. iotechnolApplBiochem, 38: 123–30.
    Youm JW, Jeon JH, Kim H, Kim YH, Ko K, Joung H. (2008). Transgenic tomatoes expressing human beta-amyloid for use as a vaccine against Alzheimer's disease. BiotechnolLett, 30: 1839–45.
    21