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A Cysteine Protease Isolated from the Latex of Ficus microcarpa: Purification and Biochemical Characterization

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Abstract

A plant protease named microcarpain was purified from the latex of Ficus microcarpa by acetonic (20–40 % saturation) precipitation, Sephadex G-75 filtration, and Mono Q-Sefinose FF chromatography. The protease was purified with a yield of 9.25 % and a purification factor of 8. The molecular weight of the microcarpain was estimated to be 20 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The purified enzyme showed maximum activity at pH 8.0 and at a temperature of 70 °C. Proteolytic activity was strongly inhibited by dithio-bis-nitrobenzoic acid (DTNB), Hg2+, and Cu2+. The N-terminal amino acid sequence of the purified microcarpain “VPETVDWRSKGAV” showed high homology with a protease from Arabidopsis thaliana. Inhibition studies and N-terminal sequence classified the enzyme as a member of the cysteine peptidases family.

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References

  1. Turk, B. (2006). Nature Reviews Drug Discovery, 5, 785–798.

    Article  CAS  Google Scholar 

  2. Frost and Sullivan. (2007). Feed enzymes: the global scenario. London, UK.

  3. Schaller, A. (2004). Planta, 220, 183–197.

    Article  CAS  Google Scholar 

  4. Boller, T. (1986), in Plant proteolytic enzymes, vol. 1: roles of proteolytic enzymes in interactions of plants with other organisms (Dalling, M.J., ed), Florida, USA, pp. 67–96.

  5. Baker, E. N., & Drenth, J. (1987). Biological macromolecules and assemblies, vol. 3. In F. Jurnak & A. McPherson (Eds.), The cysteine proteinases structure and mechanism (pp. 313–368). New York: Wiley.

    Google Scholar 

  6. Caffini, N. O., Lopez, L. M. I., Natalucci, C. L., & Priolo, N. S. (1988). Acta Farmaceutica Bonaerense, 7, 195–213.

    CAS  Google Scholar 

  7. Van der Hoorn, R. A., & Jones, J. D. (2004). Current Opinion in Plant Biology, 7, 400–407.

    Article  Google Scholar 

  8. Souza, D. P., Freitas, C. D. T., Pereira, D. A., Nogueira, F. C., Silva, F. D. A., Salas, C. E., & Ramos, M. V. (2011). Planta, 234, 183–193.

    Article  CAS  Google Scholar 

  9. Shivaprasad, H. V., Rajesh, R., Nanda, B. L., Dharmappa, K. K., & Vishwanath, B. S. (2009). Journal of Ethnopharmacology, 123, 106–109.

    Article  CAS  Google Scholar 

  10. Fonseca, K. C., Morais, N. C. G., Queiroz, M. R., Silva, M. C., Gomes, M. S., Costa, J. O., Mamede, C. C. N., Torres, F. S., Penha-Silva, N., Beletti, M. E., Canabrava, H. A. N., & Oliveiraa, F. (2010). Phytochemistry, 71, 708–715.

    Article  CAS  Google Scholar 

  11. Freitas, C. D. T., Oliveira, J. S., Miranda, M. R. A., Macedo, N. M. R., Pereira Sales, M., Villas Boas, L. A., & Ramos, M. V. (2007). Plant Physiology and Biochemistry, 45, 781–789.

    Article  CAS  Google Scholar 

  12. Vierstra, R. D. (1996). Plant Molecular Biology, 32, 275.

    Article  CAS  Google Scholar 

  13. Ervatamia. (1952), in The wealth of India, vol. 3: Council of Scientific and Industrial Research, New Delhi, pp. 192–193.

  14. Gunter, R., Hans, P. S., Friedrich, D., & Peter, L. (2002). British Journal of Haematology, 119, 1042–1051.

    Article  Google Scholar 

  15. Badgujar, S. B. (2014). Journal of Ethnopharmacology, 151, 733–739.

    Article  Google Scholar 

  16. Thankamma, L. (2003). Current Science, 84, 971–972.

    Google Scholar 

  17. Domsalla, A., & Melzig, M. F. (2008). Planta Medica, 74, 699–711.

    Article  CAS  Google Scholar 

  18. Wagner, W. L., Herbst, D. R., & Sohmer, S. H. (1999). Manual of the flowering plants of Hawaii, revised edition. 2 vols, Bishop Museum Special Publication 97. Honolulu: University of Hawaii Press and Bishop Museum Press.

    Google Scholar 

  19. Bradford, M. (1976). Analytical Biochemistry, 72, 248–254.

    Article  CAS  Google Scholar 

  20. Kembhavi, A. A., Kulkarni, A., & Pant, A. (1993). Applied Biochemistry and Biotechnology, 38, 83–92.

    Article  CAS  Google Scholar 

  21. Laemmli, U. K. (1970). Nature, 227, 680–685.

    Article  CAS  Google Scholar 

  22. Garcia-Carreno, F. L., Dimes, L. E., & Haard, N. F. (1993). Analytical Biochemistry, 214, 65–69.

    Article  CAS  Google Scholar 

  23. Shivaprasad, H. V., Rajaiah, R., Frey, B. M., Frey, F. J., & Vishwanath, B. S. (2010). Thrombosis Research, 125, 100–105.

    Article  Google Scholar 

  24. Liggieri, C., Obregon, W., Trejo, S., & Priolo, N. (2009). Acta Biochimica et Biophysica Sinca, 41, 154–162.

    Article  CAS  Google Scholar 

  25. Singh, A. N., Shukla, A. K., Jagannadham, M. V., & Dubey, V. K. (2010). Process Biochemistry, 45, 399–406.

    Article  CAS  Google Scholar 

  26. Torres, M. J., Trejo, S. A., Obregon, W. D., Aviles, F. X., Lopez, L. M. I., & Natalucci, C. L. (2012). Planta, 236, 1471–1484.

    Article  CAS  Google Scholar 

  27. Hashim, M. M., Mingsheng, D., Iqbal, M. F., & Xiaohong, C. (2011). Phytochemistry, 72, 458–464.

    Article  CAS  Google Scholar 

  28. Revell, D. F., Cummings, N. J., Baker, K. C., Collins, M. E., Taylor, M. A., Sumner, I. G., Pickersgill, R. W., Connerton, I. F., & Goodenough, P. W. (1993). Gene, 127, 221–225.

    Article  CAS  Google Scholar 

  29. Vairo Cavalli, S. E., Arribere, M. C., Cortadi, A., Caffini, N. O., & Priolo, N. S. (2003). Journal of Protein Chemistry, 22, 15–22.

    Article  CAS  Google Scholar 

  30. Morcelle del Valle, S. R., Trejo, S. A., Canals, F., Aviles, F. X., & Priolo, N. S. (2004). Protein Journal, 25, 205–215.

    Article  Google Scholar 

  31. Vairo Cavalli, S. E., Cortadi, A., Arribere, M. C., Conforti, P., Caffini, N. O., & Priolo, N. S. (2001). Biological Chemistry, 382, 879–883.

    Google Scholar 

  32. Teixeira, R. D., Ribeiro, H. A. L., Gomes, M. T. R., Lopes, M. T. P., & Salas, C. E. (2008). Plant Physiology and Biochemistry, 46, 956–961.

    Article  CAS  Google Scholar 

  33. Torres, M. J., Trejo, S. A., Natalucci, C. L., & Lopez, L. M. I. (2013). Planta, 237, 1651–1659.

    Article  CAS  Google Scholar 

  34. Vasu, P., Savary, B. J., & Cameron, R. G. (2012). Food Chemistry, 133, 366–372.

    Article  CAS  Google Scholar 

  35. Demir, Y., Güngör, A. A., Duran, E. D., & Demir, N. (2008). Food Technology and Biotechnology, 46, 286–291.

    CAS  Google Scholar 

  36. Chanda, I., Basu, S. K., Dutta, S. K., & Chanda Das, S. R. (2011). Tropical Journal of Pharmaceutical Research, 10, 705–711.

    CAS  Google Scholar 

  37. Dubey, V. K., & Jagannadham, M. V. (2003). Phytochemistry, 62, 1057–1071.

    Article  Google Scholar 

  38. Fahmy, A. S., Ali, A. A., & Mohamed, S. A. (2004). Bioresource Technology, 91, 297–304.

    Article  CAS  Google Scholar 

  39. Vallés, D., Furtado, S., & Cantera, A. M. B. (2007). Enzyme and Microbial Technology, 40, 409–413.

    Article  Google Scholar 

  40. Male, R., Lorens, L. B., Smalas, A. O., & Torrissen, K. R. (1995). European Journal of Biochemistry, 232, 677–685.

    Article  CAS  Google Scholar 

  41. Bezerra, R. S., Lins, E. J. F., Alencar, R. B., Paiva, P. M. G., Chaves, M. E. C., Coelho, L. C. B. B., & JrLB, C. (2005). Process Biochemistry, 40, 1829–1834.

    Article  CAS  Google Scholar 

  42. Siala, R., Fakhfakh, N., Hamza-Mnif, I., Nasri, M., Vallaeys, T., & Sellami-Kamoun, A. (2012). Biotechnology and Bioprocess Engineering, 17, 556–564.

    Article  CAS  Google Scholar 

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Acknowledgments

This work was funded by the Ministry of Higher Education and Scientific Research, Tunisia.

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Authors and Affiliations

  1. Laboratory of Enzyme Engineering and Microbiology, National School of Engineering of Sfax, University of Sfax, 1173-3038, Sfax, Tunisia

    Ibtissem Hamza Mnif, Rayda Siala, Rim Nasri, Samiha Mhamdi, Moncef Nasri & Alya Sellami Kamoun

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  1. Ibtissem Hamza Mnif
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  2. Rayda Siala
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  3. Rim Nasri
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  4. Samiha Mhamdi
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  5. Moncef Nasri
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Correspondence to Ibtissem Hamza Mnif.

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Mnif, I.H., Siala, R., Nasri, R. et al. A Cysteine Protease Isolated from the Latex of Ficus microcarpa: Purification and Biochemical Characterization. Appl Biochem Biotechnol 175, 1732–1744 (2015). https://doi.org/10.1007/s12010-014-1376-2

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  • DOI: https://doi.org/10.1007/s12010-014-1376-2

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