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Aqueous Two-Phase Systems for Cleanup and Recovery of Enzymes from Plants and Plant-Derived Extracts

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Protein Downstream Processing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2178))

Abstract

The increasing interest of the biopharmaceutical industry to exploit plants as a commercially viable production system is demanding the development of new strategies to maximize product recovery. Aqueous two-phase systems (ATPSs) are a primary recovery technique that has shown great potential for the efficient extraction and purification of biological products, from organelles to proteins and low-molecular-weight compounds. The evaluation of different system parameters upon the partitioning behavior can provide the conditions that favor the concentration of contaminants and the desired target protein in opposite phases. The protocols described here provide the basic strategy to explore the use of ATPSs for the isolation and partial purification of native and recombinant proteins from plants and plant-derived extracts.

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References

  1. Melnik S, Stoger E (2013) Green factories for biopharmaceuticals. Curr Med Chem 20:1038–1046

    CAS  Google Scholar 

  2. Vázquez-Villegas P, Acuña-González E, Mejía-Manzano LA, Rito-Palomares M, Aguilar O (2016) Production and optimization of a chlorophyll-free leaf protein concentrate from alfalfa (Medicago sativa) through aqueous two-phase system. Rev Mex Ing Quim 14(2):383–392

    Google Scholar 

  3. Albertsson P-A (1999) Partition of proteins in liquid polymer–polymer two-phase systems. Nature 182:709–711

    Article  Google Scholar 

  4. Benavides J, Rito-Palomares M (2008) Practical experiences from the development of aqueous two-phase processes for the recovery of high value biological products. J Chem Technol Biotechnol 83:133–142

    Article  CAS  Google Scholar 

  5. Rito-Palomares M (2004) Practical application of aqueous two-phase partition to process development for the recovery of biological products. J Chromatogr B 807:3–11

    Article  CAS  Google Scholar 

  6. McQueen L, Lai D (2019) Ionic liquid aqueous two-phase systems from a pharmaceutical perspective. Front Chem 15(7):135

    Article  CAS  Google Scholar 

  7. Hernandez-Vargas G, González-González M, Rito-Palomares M, Benavides J, González-Valdez J (2019) Thermo-separating polymer-based aqueous two-phase systems for the recovery of PEGylated lysozyme species. J Chromatogr B 1105:120–128

    Article  CAS  Google Scholar 

  8. Zhou L, Tao Zhang BJ, Li S (2019) Ultrasound-assisted aqueous two-phase extraction of resveratrol from the enzymatic hydrolysates of Polygonum cuspidatum. Food Biosci 31:100442

    Article  CAS  Google Scholar 

  9. Aguilar O, Rito-Palomares M (2010) Aqueous two-phase systems strategies for the recovery and characterization of biological products from plants. J Sci Food Agric 90:1385–1392

    Article  CAS  Google Scholar 

  10. Chow YH, Ow JL (2018) Selective recovery of amylase from pitaya (Hylocereus polyrhizus) peel using alcohol/salt aqueous two-phase system. J Eng Sci Technol 8:123–136

    Google Scholar 

  11. Gu Z, Glatz CE (2007) Aqueous two-phase extraction for protein recovery from corn extracts. J Chromatogr B 845:38–50

    Article  CAS  Google Scholar 

  12. Zhang C, Glatz CE (1999) Process engineering strategy for recombinant protein recovery from canola by cation exchange chromatography. Biotechnol Prog 15:12–18

    Article  CAS  Google Scholar 

  13. Zhang C, Love RT, Jilka JM, Glatz CE (2001) Genetic engineering strategies for purification of recombinant proteins from canola by anion exchange chromatography: an example of β-glucuronidase. Biotechnol Prog 17:161–167

    Article  CAS  Google Scholar 

  14. Platis D, Labrou NE (2006) Development of an aqueous two-phase partitioning system for fractionating therapeutic proteins from tobacco extract. J Chromatogr A 1128:114–124

    Article  CAS  Google Scholar 

  15. Aguilar O, Albiter V, Serrano-Carreon L, Rito-Palomares M (2006) Direct comparison between ion-exchange chromatography and aqueous two-phase processes for the partial purification of penicillin acylase produced by E. coli. J Chromatogr B 835:77–83

    Article  CAS  Google Scholar 

  16. Thanh VH, Shibasaki K (1976) Major proteins of soybean seeds. A straightforward fractionation and characterization. J Agric Food Chem 24:1117–1121

    Article  CAS  Google Scholar 

  17. Aguilar O, Rito-Palomares M (2008) Processing of soybean (Glycine max) extracts in aqueous two-phase systems as a first step for the potential recovery of recombinant proteins. J Chem Technol Biotechnol 83:286–293

    Article  CAS  Google Scholar 

  18. Sandhya C, Sumantha A, Szakacs G, Pandey A (2005) Comparative evaluation of neutral protease production by Aspergillus oryzae in submerged and solid-state fermentation. Process Biochem 40(8):2689–2694

    Article  CAS  Google Scholar 

  19. Aguilar O, Glatz CE, Rito-Palomares M (2009) Characterization of green-tissue protein extract from alfalfa (Medicago sativa) exploiting a 3-D technique. J Sep Sci 32:3223–3231

    Article  CAS  Google Scholar 

  20. Ibarra-Herrera CC, Aguilar O, Rito-Palomares M (2011) Application of an aqueous two-phase systems strategy for the potential recovery of a recombinant protein from alfalfa (Medicago sativa). Sep Purif Technol 77:94–98

    Article  CAS  Google Scholar 

  21. Zaslavsky A (1995) Aqueous two-phase partitioning. Physical chemistry and bioanalytical applications. Marcel Dekker Inc., New York, NY

    Google Scholar 

  22. Ooi CW, Tey BT, Hii SL, Kamal SMM, Lan JCW, Ariff A, Ling TC (2009) Purification of lipase derived from Burkholderia pseudomallei with alcohol/salt-based aqueous two-phase systems. Process Biochem 44(10):1083–1087

    Article  CAS  Google Scholar 

  23. Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  24. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  25. AOAC International (2016) Official methods of analysis of AOAC international, 19th edn. AOAC International, Rockville, MD

    Google Scholar 

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Acknowledgments

The authors wish to acknowledge the financial support from Tecnológico de Monterrey Bioprocesses Research Chair (Grant 0020209I13).

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Authors and Affiliations

  1. Tecnologico de Monterrey, Escuela de Ingenieria y Ciencias, Monterrey, Mexico

    Oscar Aguilar, Erick Heredia-Olea & Esther Perez-Carrillo

  2. Tecnologico de Monterrey, Escuela de Medicina y Ciencias de la Salud, Monterrey, Mexico

    Marco Rito-Palomares

Authors
  1. Oscar Aguilar
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  2. Erick Heredia-Olea
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  3. Esther Perez-Carrillo
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  4. Marco Rito-Palomares
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Corresponding author

Correspondence to Marco Rito-Palomares .

Editor information

Editors and Affiliations

  1. Laboratory of Enzyme Technology, Department of Biotechnology, School of Applied Biology and Biotechnology, Agricultural University of Athens, Athens, Greece

    Nikolaos E. Labrou

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Aguilar, O., Heredia-Olea, E., Perez-Carrillo, E., Rito-Palomares, M. (2021). Aqueous Two-Phase Systems for Cleanup and Recovery of Enzymes from Plants and Plant-Derived Extracts. In: Labrou, N.E. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 2178. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0775-6_7

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  • DOI: https://doi.org/10.1007/978-1-0716-0775-6_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0774-9

  • Online ISBN: 978-1-0716-0775-6

  • eBook Packages: Springer Protocols

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