Skip to main content
Log in

Thermal Characterization and Cytotoxicity of Complexes Formed by Papain and Cyclodextrin

  • Original Paper
  • Published:
Journal of Biological Physics Aims and scope Submit manuscript

Abstract

Papain is a proteolytic enzyme with restricted applications due to its limited stability. Cyclodextrins are widely used in pharmaceutical formulations once they are able to form complexes with other molecules, improving their stability and bioavailability. The purpose of the present paper was to analyze complexes formed by papain/hydroxypropyl-β-cyclodextrin and papain/β-cyclodextrin by thermal analysis and cytotoxicity tests to verify their possible interactions and toxicological behavior. Complex solutions were prepared at different molar ratios and collected as a function of time to be lyophilized and analyzed. Results showed changes in endothermic events and cytotoxicity profiles. A complex formation for both complexes is observed; nevertheless, β-cyclodextrin was able to change the enzyme characteristics more efficiently.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Sim, Y.C., Lee, S.G., Lee, D.C., Kang, B.Y., Park, K.M., Lee, J.Y., Kim, M.S., Chang, I.S., Rhee, J.S.: Stabilization of papain and lysozyme for application to cosmetic products. Biotechnol. Lett. 22, 137–140 (2000). doi:10.1023/A:1005670323912

    Article  Google Scholar 

  2. Kamphuis, I.G., Kalk, K.H., Swarte, M.B.A., Drent, H.J.: Structure of papain refined at 1.65 angstrom resolution. J. Mol. Biol. 179, 233–256 (1984). doi:10.1016/0022–2836(84)90467-4

    Article  Google Scholar 

  3. O’Neil, M.J.: The Merck Index, 13th edn. Merck Research Laboratories, Whitehouse Station, NJ (2001)

    Google Scholar 

  4. Sathish, H.A., Kumar, P.R., Prakash, V.: Mechanism of solvent induced thermal stabilization of papain. Int. J. Biol. Macromol. 41, 383–390 (2007). doi:10.1016/j.ijbiomac.2007.05.009

    Article  Google Scholar 

  5. Zhuang, P., Butterfield, D.A.: Denaturation studies of active-site labeled papain using electron paramagnetic resonance and fluorescence spectroscopy. Biophys. J. 60, 623–628 (1991)

    Article  Google Scholar 

  6. Rialdi, G., Battistel, E.: Unfolding mechanism and stability of immobilized papain. J. Therm. Anal. 47, 17–25 (1996). doi:10.1007/BF01982682

    Article  Google Scholar 

  7. Legendre, J.Y., Rault, I., Petit, A., Luijten, W., Demuynck, I., Horvath, S., Ginot, Y.M., Cuine, A.: Effects of β-cyclodextrins on the skin: implications for the transdermal delivery of piribedil and a novel cognition enhancing-drug, S-9977. Eur. J. Pharm. Sci. 3, 311–322 (1995). doi:10.1016/0928-0987(95)00020-0

    Article  Google Scholar 

  8. Singh, M., Sharma, R., Banerjee, U.C.: Biotechnological applications of cyclodextrins. Biotechnol. Adv. 20, 341–359 (2002). doi:10.1016/S0734-9750(02)00020-4

    Article  Google Scholar 

  9. Sumner, I.G., Harris, G.W., Taylor, M.A.J., Pickersgill, R.W., Owen, A.J., Goodenough, W.: Factors effecting the thermostability of cysteine proteinases from Carica papaya. Eur. J. Biochem. 214, 129–134 (1993). doi:10.1111/j.1432-1033.1993.tb17904.x

    Article  Google Scholar 

  10. Rialdi, G., Battistel, E.: Thermodynamics of proteins in unusual environments. Biophys. Chem. 126, 65–79 (2006). doi:10.1016/j.bpc.2006.05.029

    Article  Google Scholar 

  11. Pollizi, K.M., Bommarius, A.S., Broering, J.M., Chaparro-Riggers, J.F.: Stability of biocatalysts. Curr. Opin. Chem. Biol. 11, 220–225 (2007). doi:10.1016/j.cbpa.2007.01.685

    Article  Google Scholar 

  12. Shnyrov, V.L., Sanchez-Ruiz, J.M., Boiko, B.N., Zhadan, G.G., Permyakov, E.A.: Applications of scanning microcalorimetry in biophysics and biochemistry. Thermochim. Acta 302, 165–180 (1997). doi:10.1016/S0040-6031(97)00238-4

    Article  Google Scholar 

  13. National Institutes of Health: Guidance document on using in vitro data to estimate in vivo starting doses for acute toxicity. No. 01-4500. http://www.epa.gov/chemrtk/pubs/general/nih2001b.pdf, August (2001)

  14. Freshney, R.I.: Culture of Animal Cells: A Manual of Basic Technique, 5th edn. Wiley-Liss, New York (2005)

    Google Scholar 

  15. Hernández-Arana, A., Soriano-Garcia, M.: Detection and characterization by circular dichroism of a stable intermediate state formed in the thermal unfolding of papain. Biochim. Biophys. Acta 954, 170–175 (1988)

    Google Scholar 

  16. Fernández, M., Fragoso, A., Cao, R., Villalonga, R.: Stabilization of α-chymotrypsin by chemical modification with monoamine cyclodextrin. Process Biochem. 40, 2091–2094 (2005). doi:10.1016/j.procbio.2004.07.023

    Article  Google Scholar 

  17. Lee, J.C., Timasheff, S.N.: The stabilization of proteins by sucrose. J. Biol. Chem. 256, 7193–7201 (1981)

    Google Scholar 

  18. Arakawa, T., Timasheff, S.N.: Stabilization of protein structure by sugars. Biochemistry 21, 6536–6544 (1982). doi:10.1021/bi00268a033

    Article  Google Scholar 

  19. Martin Del Valle, E.M.: Cyclodextrins and their uses: a review. Process Biochem. 39, 1033–1046 (2004). doi:10.1016/S0032–9592(03)00258-9

    Article  Google Scholar 

  20. Schneiderman, E., Stalcup, A.M.: Cyclodextrins: a versatile tool in separation science. J. Chromatogr. B 745, 83–102 (2000). doi:10.1016/S0378-4347(00)00057-8

    Article  Google Scholar 

  21. Szetjli, J.: Introduction and general overview of cyclodextrin chemistry. Chem. Rev. 98, 1743–1753 (1998). doi:10.1021/cr970022c

    Article  Google Scholar 

  22. Promega: Technical Bulletin no. 169: Cell titer 96® aqueous non-radiactive cell proliferation assay. Updated 12/2006. http://www.promega.com/enotes/applicattions/sap0017_print.htm. Cited 5 Dec 2005 (2005)

  23. Moraes, C.M., Araujo, D.R., Issa, M.G., Ferraz, H.G., Yokaichiya, F., Franco, M.K.K.D., Mazzaro, I., Lopes, P.S., Gonçalves, M.M., de Paula, E., Fraceto, L.F.: Inclusion complex of S(-) bupivacaine and 2-hydroxypropyl-β-cyclodextrin: study of morphology and citotoxicity. Rev. Ciênc. Farm. Basica Apl. 27(3), 207–212 (2006)

    Google Scholar 

  24. Lopes, P.S.: Avaliação da eficácia e segurança da papaína como promotor de absorção cutâneo utilizando técnicas biofísicas e cultura celular de queratinócitos humanos. Ph.D. thesis, University of São Paulo (2003)

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Patricia S. Lopes.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Varca, G.H.C., Andréo-Filho, N., Fraceto, L.F. et al. Thermal Characterization and Cytotoxicity of Complexes Formed by Papain and Cyclodextrin. J Biol Phys 33, 463–475 (2007). https://doi.org/10.1007/s10867-008-9098-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10867-008-9098-8

Keywords

Navigation