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Isolation of papain from ripe papaya peel using aqueous two-phase extraction

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Abstract

Papaya peel is a good source of papain enzyme (cysteine protease) but currently unutilized for any commercial purpose and disposed of as waste, thereby becoming a source of pollution. Therefore, this study investigates the behavior of papain enzyme (cysteine protease) in an aqueous two-phase system (ATPS) comprising polymer/salt. Considering the excluded volume effect, polyethylene glycol (PEG) with a molecular weight of 6000 g/mol was used to form the ATPS. Sulphate salts (ammonium sulphate and sodium sulphate) were chosen to form phases owing to their ability to promote the hydrophobic difference between the phases. From the result, the best ATPS condition for the partitioning of cysteine protease was 10% (w/w) PEG 6000 + 18% (w/w) Na2SO4 at constant pH 9 and obtained the highest enzyme activity (Ke) 1.43 which increased the purity by 4.08-fold (PF) with the yield of 26.38%. PEG/salt ATPS has been shown to work well to purify partitioned cysteine protease, which can be scaled up and extracted with a high yield and purity factor. In addition, the paper studies the (Sodium dodecyl-sulfate polyacrylamide gel electrophoresis) SDS-PAGE patterns of cysteine protease partitioned from papaya peels. The ATPS system is therefore effective in recovering and purifying papain enzymes from papaya peel.

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References

  1. W. Aehle, Enzymes in Industry: Production and Application (Wiley-VCH, New York, 2004)

    Google Scholar 

  2. R. Tomar, R. Kumar, M.V. Jagannadham, J. Agric, Food Chem. 56(4), 1479–1487 (2008). https://doi.org/10.1021/jf0726536

    Article  CAS  Google Scholar 

  3. A. Sumantha, C. Larroche, A. Pandey, Food Technol. Biotechnol. 44, 211–220 (2006)

    CAS  Google Scholar 

  4. M. Singla, A. Singh, N. Sit, J. Food Process. Eng. e14119 (2022). https://doi.org/10.1111/jfpe.4119

    Article  Google Scholar 

  5. V. Yogiraj, P.K. Goyal, C.S. Chauhan, A. Goyal, B. Vyas, Int. J. Herb. Med. 2(5), 01–08 (2014)

    Google Scholar 

  6. FAOSTAT (2021). http://www.fao.org/faostat/en/#data/QC

  7. P.D. Pathak, S.A. Mandavgane, B.D. Kulkarni, Waste Biomass Valorization 10(6), 1755–1766 (2019). https://doi.org/10.1007/s12649-017-0181-x

    Article  Google Scholar 

  8. Y.Q. Ling, H.L. Nie, S.N. Su, C. Branford-White, L.M. Zhu, Sep. Purif. Technol. 73(3), 343–348 (2010). https://doi.org/10.1016/j.seppur.2010.04.020

    Article  CAS  Google Scholar 

  9. M. Zhang, P. Hu, Q. Liang, H. Yang, Q. Liu, Y. Wang, G. Luo, J. Chromatogr. B 858(1–2), 220–226 (2007). https://doi.org/10.1016/j.jchromb.2007.08.041

    Article  CAS  Google Scholar 

  10. R. Gupta, Q. Beg, P. Lorenz, Appl. Microbiol. Biotechnol. 59(1), 15–32 (2002). https://doi.org/10.1007/s00253-002-0975-y

    Article  CAS  PubMed  Google Scholar 

  11. J.P. Chen, M.S. Lee, Enzyme Microb. Technol. 17(11), 1021–1027 (1995). https://doi.org/10.1016/0141-0229(95)00030-5

    Article  CAS  Google Scholar 

  12. E. Andersson, B. Hahn-Hägerdal, Enzyme Microb. Technol. 12(4), 242–254 (1990). https://doi.org/10.1016/0141-0229(90)90095-8

    Article  CAS  PubMed  Google Scholar 

  13. T. Tianwei, H. Qing, L. Qiang, Biotechnol. Lett. 24(17), 1417–1420 (2002). https://doi.org/10.1023/A:1019850531640

    Article  Google Scholar 

  14. R. Hatti-Kaul, Mol. Biotechnol. 19(3), 269–277 (2001). https://doi.org/10.1385/MB:19:3:269

    Article  CAS  PubMed  Google Scholar 

  15. H. Hustedt, K.H. Kroner, N. Papamichael, Process. Biochem. 23(5), 129–137 (1988)

    CAS  Google Scholar 

  16. A. Veide, A.L. Smeds, S.O. Enfors, Biotechnol. Bioeng. 25(7), 1789–1800 (1983). https://doi.org/10.1002/bit.260250709

    Article  CAS  PubMed  Google Scholar 

  17. M.R. Kula, K.H. Kroner, H. Hustedt, H. Schutte, Enzym Eng. 69–74 (1982). https://doi.org/10.1007/978-1-4615-9290-7_6

  18. U. Menge, in Aqueous two-phase systems: methods and protocols, ed. by R. Hatti-Kaul (Humana Press, Totowa, 2000) pp. 235–249

  19. P. Albertsson, Partition of cell Particles and Macromolecules, 3rd edn. (Wiley, New York, 1986)

    Google Scholar 

  20. A. Venâncio, C. Almeida, J. Teixeira, J. Chromatogr, B Biomed. Appl. 680(1–2), 131–136 (1996). https://doi.org/10.1016/0378-4347(95)00419-X

    Article  Google Scholar 

  21. H. Hustedt, K.H. Kroner, M.R. Kula, in Partitioning in Aqueous Two-Phase Systems; Theory, Methods, Uses and Application in Biotechnology, ed. by H. Walter, D. E. Brooks, D. Fisher, (Academic press, Orlando, 1985) pp. 529–587

  22. J. Vernau, M.R. Kula, Biotechnol. Appl. Biochem. 12(4), 397–404 (1990)

    CAS  Google Scholar 

  23. M.M. Bradford, Anal. Biochem. 72(1–2), 248–254 (1976). https://doi.org/10.1006/abio.1976.9999

    Article  CAS  PubMed  Google Scholar 

  24. U.K. Laemmli, Nature 227, 680–685 (1970). https://doi.org/10.1038/227680a0

    Article  CAS  PubMed  Google Scholar 

  25. A.L. Grilo, M. Raquel Aires-Barros, A.M. Azevedo, Sep. Purif. Rev. 45(1), 68–80 (2016). https://doi.org/10.1080/15422119.2014.983128

    Article  Google Scholar 

  26. E.L. Smith, J.R. Kimmel, D.M. Brown, J. Biol. Chem. 207(2), 533–549 (1954). https://doi.org/10.1016/S0021-9258(18)65670-4

    Article  CAS  PubMed  Google Scholar 

  27. J.A. Asenjo, B.A. Andrews, J. Chromatogr. A 1218(49), 8826–8835 (2011). https://doi.org/10.1016/j.chroma.2011.06.051

    Article  CAS  PubMed  Google Scholar 

  28. J.C. Marcos, L.P. Fonseca, M.T. Ramalho, J.M.S. Cabral, J. Chromatogr, B Biomed. Appl. B 711, 295–299 (1998). https://doi.org/10.1016/S0378-4347(97)00633-6

    Article  CAS  Google Scholar 

  29. M.A. Eiteman, J.L. Gainer, Biochim. Biophys. Acta Gen. Subj. 1073(3), 451–455 (1991). https://doi.org/10.1016/0304-4165(91)90214-2

    Article  CAS  Google Scholar 

  30. T. Arakawa, L.O. Narhi, Biotechnol. Appl. Biochem. 13(2), 151–172 (1991)

    CAS  PubMed  Google Scholar 

  31. T. Arakawa, S.N. Timasheff, Biochemistry 23(25), 5912–5923 (1984). https://doi.org/10.1021/bi00320a004

    Article  CAS  PubMed  Google Scholar 

  32. B.Y. Zaslavsky, Aqueous two-phase Partitioning: Physical Chemistry and Bioanalytical Applications (CRC press,, New York, 1994)

    Google Scholar 

  33. A. Salabat, M.H. Abnosi, A.R. Bahar, J. Chromatogr. B 858, 234–238 (2007). https://doi.org/10.1016/j.jchromb.2007.08.039

    Article  CAS  Google Scholar 

  34. H. Yue, Q. Yuan, W. Wang, Biochem. Eng. J. 37(3), 231–237 (2007). https://doi.org/10.1016/j.bej.2007.05.002

    Article  CAS  Google Scholar 

  35. G. Johansson, Mol. Cell. Biochem. 4(3), 169–180 (1974). https://doi.org/10.1007/BF01731478

    Article  CAS  PubMed  Google Scholar 

  36. S. Nalinanon, S. Benjakul, W. Visessanguan, H. Kishimura, Process. Biochem. 44(4), 471–476 (2009). https://doi.org/10.1016/j.procbio.2008.12.018

    Article  CAS  Google Scholar 

  37. R. Tapadar, M. Nandi, R. Chatterjee, K. Mukherjee, K. Majumder, J. BioSci. Biotechnol. 8(2), 168–173 (2019)

    Google Scholar 

  38. M. de H.C. Maciel, C.A. Ottoni, P.N. Herculano, T.S. Porto, A.L. Porto, C. Santos, L. Nelson, A.M. Keila, C. Souza-Motta, Fluid Ph. Equilibria 371, 125–130 (2014). https://doi.org/10.1016/j.fluid.2014.03.018

    Article  CAS  Google Scholar 

  39. M.A. Fakhari, F. Rahimpour, M. Taran, J. Chromatogr. B 1063, 1–10 (2017). https://doi.org/10.1016/j.jchromb.2017.08.007

    Article  CAS  Google Scholar 

  40. K. Naganagouda, V.H. Mulimani, Process. Biochem. 43(11), 1293–1299 (2008). https://doi.org/10.1016/j.procbio.2008.07.016

    Article  CAS  Google Scholar 

  41. J.N. Baskir, T.A. Hatton, U.W. Suter, Biotechnol. Bioeng. 34(4), 541–558 (1989). https://doi.org/10.1002/bit.260340414

    Article  CAS  PubMed  Google Scholar 

  42. T. Vernet, P.J. Berti, C. de Montigny, R. Musil, D.C. Tessier, R. Ménard, M.C. Magny, A.C. Storer, D.Y. Thomas, J. Biol. Chem. 270(18), 10838–10846 (1995). https://doi.org/10.1074/jbc.270.18.10838

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors would like to acknowledge the fellowship received from All India Council of Technical Education (AICTE), New Delhi, India for carrying out the work. The authors would also like to acknowledge the help received from Prof. Robin Doley and Ms. Nyumpi Bagra for using the facilities of their lab and helping in analysis of the samples.

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No external funding was received to carry out the present work.

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Authors and Affiliations

  1. Department of Food Engineering and Technology, Tezpur University, 784028, Assam, India

    Mohit Singla & Nandan Sit

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  1. Mohit Singla
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  2. Nandan Sit
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Contributions

Mohit Singla: Methodology, validation, Formal analysis, investigation, Writing-original draft; Nandan Sit: Conceptualization, validation, writing-review and editing, visualization, supervision, project administration.

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Correspondence to Nandan Sit.

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Singla, M., Sit, N. Isolation of papain from ripe papaya peel using aqueous two-phase extraction. Food Measure 17, 1685–1692 (2023). https://doi.org/10.1007/s11694-022-01741-3

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