Skip to main content
SpringerLink
Log in

Proteolysis of mesophilic and thermophilic α-amylases

A comparative study

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

A comparative study was performed on limited and extensive proteolysis of mesophilic (from Bacillus amyloliquefaciens [BAA]) and thermophilic (from Bacillus licheniformis [BLA]) α-amylases using trypsin. As expected, the thermophilic enzyme showed greater resistance to digestion by the protease. While the catalytic potential of BLA was enhanced on proteolysis, that of BAA was diminished owing to this process. Combined with greater catalytic activity, a lower thermal stability was observed for BLA on proteolytic treatment. For both enzymes, the extent of proteolytic cleavage was reduced in the presence of various stabilizing agents. The digestion patterns are explained in terms of available information in the literature on the structure of these proteins, especially in relation to segmental mobility.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Vieille, C., Burdette, D. S., and Zeikus, J. G. (1996), Biotechnol. Annu. Rev. 2, 1–79.

    CAS  Google Scholar 

  2. Ludlow, J. M. and Clark, D. S. (1991), CRC Crit. Rev. Biotechnol. 10, 321–345.

    Article  CAS  Google Scholar 

  3. Clark, D. and Kelly, R. (1990), Chemtech 20, 654–662.

    CAS  Google Scholar 

  4. Niehaus, F., Bertoldo, C., Kähler, M., and Antranikian, G. (1999), Appl. Microbiol. Biotechnol. 51, 711–729.

    Article  CAS  Google Scholar 

  5. Daniel, R. M., Cowan, D. A., Morgan, H. W., and Curran, M. P. (1982), Biochem. J. 207, 641–644.

    CAS  Google Scholar 

  6. Veronese, F. M., Boccu, E., Schiaron, O., Grandi, C., and Fontana, A. (1984), J. Appl. Biochem. 6, 39–47.

    CAS  Google Scholar 

  7. Fontana, A., Fassina, G., Vite, C., Dalzoppo, D., Zamai, M., and Zambonin, M. (1986), Biochemistry 25, 1847–1851.

    Article  CAS  Google Scholar 

  8. Neurath, H. (1980), in Protein Folding, Jaenicke, R., ed., Elsevier/North-Holland, Amsterdam, pp. 501–524.

    Google Scholar 

  9. Polverino de Laureto, P., Scaramella, E., De Filippis, V., Bruix, M., Rico, M., and Fontana, A. (1997), Protein Sci. 6, 860–872.

    Article  CAS  Google Scholar 

  10. Richards, F. M. and Withayathil, P. J. (1959), J. Biol. Chem. 234, 1459–1464.

    CAS  Google Scholar 

  11. Bennett, W. S. and Huber, R. (1984), CRC Crit. Rev. Biochem. 15, 291–384.

    CAS  Google Scholar 

  12. Frauenfelder, H., Petsko, G. A., and Tsernoglou, D. (1979), Nature (Lond.) 280, 588–593.

    Article  Google Scholar 

  13. Artymiuk, P. J., Blake, C. C. F., Grace, D. E. P., Oatley, S. J., Philips, D. C., and Sternberg, M. J. E. (1979), Nature (Lond.) 280, 563–568.

    Article  CAS  Google Scholar 

  14. Sternberg, M. J. E., Grace, D. E. P., and Philips, D. C. (1979), J. Mol. Biol. 130, 231–253.

    Article  CAS  Google Scholar 

  15. Machius, M., Wiegand, G., and Huber, R. (1995), J. Mol. Biol. 246, 545–559.

    Article  CAS  Google Scholar 

  16. Vihinen, M. and Mäntsälä, P. (1989), Crit. Rev. Biochem. Mol. Biol. 24, 329–418.

    Article  CAS  Google Scholar 

  17. Morren, M. A., Janssens, V., Dooms-Goossens, A., Van Hoeyveld, E., Cornelis, A., De Wolf-Peeters, C., and Heremans, A. (1993), J. Am. Acad. Dermatol. 29, 723–728.

    Article  CAS  Google Scholar 

  18. Khajeh, K. and Nemat-Gorgani, M. (2001), Appl. Biochem. Biotechnol. 90, 47–55.

    Article  CAS  Google Scholar 

  19. Millero, F. J. (1972), in Water and Aqueous Solution, Horne, R. A., ed., John Wiley & Sons, New York, pp. 519–595.

    Google Scholar 

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

    Article  CAS  Google Scholar 

  21. Haegele, E. O., Schaich, E., Rauscher, E., Lehmann, P., Bürk, H., and Wahlefeld, A. W. (1982), Clin. Chem. 28, 2201–2205.

    CAS  Google Scholar 

  22. Fritz, H. (1984), in Methods of Enzymatic Analysis, 3rd ed., vol. 5, Bergmeyer, H. U., ed., Verlag Chemie, Weinheim, pp. 1802–1806.

    Google Scholar 

  23. Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951), J. Biol. Chem. 193, 265–275.

    CAS  Google Scholar 

  24. Dixon, H. B. F. and Perham, R. N. (1968), Biochem. J. 109, 312–314.

    CAS  Google Scholar 

  25. Scandurra, R., Consalvi, V., Chiaraluce, R., Poiliti, L., and Engel, P. C. (1998), Biochimi 80, 933–941.

    Article  CAS  Google Scholar 

  26. Stetter, K. O. (1999), FEBS Lett. 452, 22–25.

    Article  CAS  Google Scholar 

  27. Haney, P. J., Badger, J. H., Buldak, G. L., Reich, C. I., Woese, C. R., and Olsen, G. J. (1999), Proc. Natl. Acad. Sci. USA 96, 3578–3583.

    Article  CAS  Google Scholar 

  28. JaneČek, Š and BaláŽ, Š (1992), FEBS Lett. 304, 1–3.

    Article  Google Scholar 

  29. JaneČek, Š (1993), Int. J. Biol. Macromol. 15, 317, 318.

    Article  Google Scholar 

  30. Tomazic, S. J. and Klibanov, A. (1988), J. Biol. Chem. 263, 3086–3091.

    CAS  Google Scholar 

  31. Tomazic, S. J. and Klibanov, A. (1988), J. Biol. Chem. 263, 3092–3096.

    CAS  Google Scholar 

  32. Violet, M. and Meunier, J. C. (1989), Biochem. J. 263, 665–670.

    CAS  Google Scholar 

  33. Brosnan, M. P., Kelly, C. T., and Fogarty, W. M. (1992), Eur. J. Biochem. 203, 225–231.

    Article  CAS  Google Scholar 

  34. Suzuki, Y., Ito, N., Yuuki, T., Yamagata, H., and Udaka, S. (1989), J. Biol. Chem. 264, 18,933–18,938.

    CAS  Google Scholar 

  35. Sundaram, T. K., Wright, I. P., and Wilkinson, A. E. (1980), Biochemistry 19, 2017–2022.

    Article  CAS  Google Scholar 

  36. Fujita, S. C., Oshima, T., and Imahori, K. (1976), Eur. J. Biochem. 64, 57–68.

    Article  CAS  Google Scholar 

  37. Závodszky, P., Kardos, J., Svingor, A., and Petsko, G. A. (1998), Proc. Natl. Acad. Sci. USA 95, 7406–7411.

    Article  Google Scholar 

  38. Klibanov, A. M. (1997), Trends Biotechnol. 15, 97–101.

    Article  CAS  Google Scholar 

  39. Griebenow, K., Laureano, Y. D., Santos, A. M., Clemente, I. M., Rodriguez, L., Vidal, M. W., and Barletta, G. (1999), J. Am. Chem. Soc. 121, 8157–8163.

    Article  CAS  Google Scholar 

  40. Yuuki, T., Nomura, T., Tezuka, H., Tsuboi, A., Yamagata, H., Tsukagoshi, N., and Udaka, S. (1985), J. Biochem. 98, 1147–1156.

    CAS  Google Scholar 

  41. Craik, C. S., Largman, C., Fletcher, T., Barr, P., Fletterick, R., and Rutter, W. J. (1985), Science 228, 291–297.

    Article  CAS  Google Scholar 

  42. Perona, J. J. and Craik, C. S. (1995), Protein Sci. 4, 337–360.

    Article  CAS  Google Scholar 

  43. Branden, C. and Tooze, J. (1999), in Introduction to Protein Structure, 2nd ed., Garland, New York, pp. 13–34.

    Google Scholar 

  44. Morea, V., Leplae, R., and Tramontano, A. (1998), Biotech. Annu. Rev. 4, 177–214.

    Article  CAS  Google Scholar 

  45. Barrett, A. J., Rawlings, N. D., and Woessner, J. F. (1998), in Handbook of Proteolytic Enzymes, Academic Press, pp. 12–21.

  46. De Cordet, S., Hendrickx, M., Maesmans, G., and Tobback, P. (1994), Biotechnol. Bioeng. 43, 107–114.

    Article  Google Scholar 

  47. Timasheff, S. N. (1993), Annu. Rev. Biophys. Biomol. Struct. 22, 67–97.

    Article  CAS  Google Scholar 

  48. Arakawa, T., Bhat, R., and Timasheff, S. N. (1990), Biochemistry 19, 1914–1923.

    Article  Google Scholar 

  49. Arakawa, T., Bhat, R., and Timasheff, S. N. (1990), Biochemistry 19, 1924–1931.

    Article  Google Scholar 

  50. Arakawa, T. and Timasheff, S. N. (1984), Biochemistry 23, 5912–5923.

    Article  CAS  Google Scholar 

  51. Richards, F. M. (1977), Annu. Rev. Biophys. Bioeng. 6, 151–175.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Biochemistry and Biophysics, University of Tehran, PO Box 13145-1384, Tehran, Iran

    Khosro Khajeh, Sadighe Khezre-Barati & Mohsen Nemat-Gorgani

Authors
  1. Khosro Khajeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sadighe Khezre-Barati
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mohsen Nemat-Gorgani
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mohsen Nemat-Gorgani.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Khajeh, K., Khezre-Barati, S. & Nemat-Gorgani, M. Proteolysis of mesophilic and thermophilic α-amylases. Appl Biochem Biotechnol 94, 97–109 (2001). https://doi.org/10.1385/ABAB:94:2:097

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1385/ABAB:94:2:097

Index Entries

Search

Navigation