Skip to main content
SpringerLink
Log in

Microbial synthesis of chitinase in solid cultures and its potential as a biocontrol agent against phytopathogenic fungus Colletotrichum gloeosporioides

  • Original Research Articles
  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Antifungal activity of chitinase can be effectively utilized in biologic pest control strategies. Because solid-state cultivation has been termed a cost-effective means for fungal growth and metabolite production, chitinase production by Trichoderma harzianum was studied using wheat bran-based solid medium containing 1% colloidal chitin. Chitinase synthesis was found to be growth associated because maximum enzyme (5.4 U/g of dry substrate) and biomass production occurred at 72h. Substrate moisture had a critical impact on chitinase production; five grams of medium having an initial moisture content of 68.4% when incubated for 72 h increased the enzyme yield to 9.3 U/g of dry substrate. Optimization of colloidal chitin concentration showed that improvements in chitinase yield and maximum activity were attained with a 2% (w/w) concentration. Supplementation of additional nitrogen sources also influenced enzyme production, and the best yield was obtained with yeast extract. The effect of crude chitinase on hyphal morphology of the phytopathogenic fungus Collelotrichum gloeosporioides was swelling as well as lysis of hyphal wall, depending on the age of the mycelium. Studies of pH and thermal stability showed that crude culture filtrate was active over pH 4.0–6.0 and retained about 48.2% activity after 40 min of incubation at 40°C.

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. Jackson, M. A. (1997), J. Ind. Microbiol. 19, 180–187.

    Article  CAS  Google Scholar 

  2. Jeuniaux, C. (1996), in Methods in Enzymology, vol. 8, Neufield, E. F. and Ginsburg, V., eds., Academic, New York, pp. 644–650.

    Google Scholar 

  3. Sandhya, C., Binod, P., and Pandey, A. (2004), in Management of Fruits and Vegetable Diseases, Chincholkar, S. B., ed., Elsevier, UK, in press.

    Google Scholar 

  4. Mahadevan, B. and Crawford, D. L. (1997), Enzyme Microb. Technol. 20, 489–493.

    Article  CAS  Google Scholar 

  5. Pandey, A. (1992), Process Biochem. 27, 109–117.

    Article  CAS  Google Scholar 

  6. Pandey, A., Selvakumar, P., Soccol, C. R., and Nigam, P. (1999), Curr. Sci. 77, 149–162.

    CAS  Google Scholar 

  7. Pandey, A., Soccol, C. R., and Mitchell, D. A. (2000), Process Biochem. 35, 1153–1169.

    Article  CAS  Google Scholar 

  8. Pandey, A. and Soccol, C. R. (2000), J. Sci. Ind. Res. 59, 12–22.

    CAS  Google Scholar 

  9. Pandey, A., Soccol, C. R., Rodriguez-leon, J. A., and Poonam, N. (2001), in Solid State Fermentation in Biotechnology Fundamentals and Application, Asiatech, New Delhi, pp. 17–18.

    Google Scholar 

  10. Matsumoto, Y., Revah, S., Saucedo, G., Hall, G. M., and Shirai, K. (2001), in Chitin Enzymology, Muzzarelli, R. A. A., ed., Atec, Italy, pp. 381–389.

    Google Scholar 

  11. Rattanakit, N., Plikomol, A., Yano, S., Wakayama, M., and Tachiki, T. (2002), J. Biosci. Bioeng. 93, 550–556.

    CAS  Google Scholar 

  12. Nampoothiri, K. M., Baiju, T. V., Sandhya, C., Sabu, A., Szakacs, G., and Pandey, A. (2004), Process Biochem. 39, 1583–1590.

    Article  CAS  Google Scholar 

  13. Krishnamurthy, J., Samiyappan, R., Vidhyasekharan, P., Nakkeeran, S., Rajeswari, E., Raja, J. A. J., and Balasubramanian, P. (1999), J. Biosci. 24, 207–213.

    Article  Google Scholar 

  14. Cherif, M. and Benhamou, N. (1990), Phytopathology 80, 1406–1414.

    CAS  Google Scholar 

  15. Chet, I. (1987), in Innovative Approaches to Plant Disease Control, Chet, I., ed., Wiley, New York, pp. 137–160.

    Google Scholar 

  16. Papavizas, G. C. (1985), Annu. Rev. Phytopathol. 23, 23–54.

    Article  Google Scholar 

  17. Sandhya, C., Adapa, L. K., Nampoothiri, K. M., Binod, P., Szakacs, G., and Pandey, A. (2004), J. Basic Microbiol. 44, 49–58.

    Article  CAS  Google Scholar 

  18. Miller, G. L. (1959), Anal. Chem. 31, 426–428.

    Article  CAS  Google Scholar 

  19. Sakurai, Y., Lee, T. H., and Shiota, H. (1997), Agric. Biol. Chem. 41, 619–624.

    Google Scholar 

  20. Suresh, P. V. and Chandrasekharan, M. (1999), Process Biochem. 34, 257–267.

    Article  CAS  Google Scholar 

  21. Pandey, A. (1990), Biol. Wastes 34, 11–19.

    Article  CAS  Google Scholar 

  22. Ramachandran, S., Patel, A. K., Nampoothiri, K. M., Sandhya, C., Szakacs, G., Soccol, C. R., and Pandey, A. (2004), Braz. Arch. Biol. Technol. 47, 3.

    Article  Google Scholar 

  23. Sabu, A., Sarita, S., Pandey, A., Bogar, B., Szakacs, G., and Soccol, C. R. (2002), Appl. Biochem. Biotechnol. 102, 251–260.

    Article  Google Scholar 

  24. Desgranges, C., Vergoignan, C., Georges, M., and Durand, A. (1991), Appl. Microbiol. Biotechnol. 35, 200–205.

    CAS  Google Scholar 

  25. Vaidya, R. J., Shah, I. M., Vyas, P. R., and Chhatpar, H. S. (2001), World J. Microbiol. Biotechnol. 17, 691–696.

    Article  CAS  Google Scholar 

  26. Nishio, N., Tai, K., and Nagai, S. (1979), Eur. J. Appl. Microbiol. Biotechnol. 8, 263–270.

    Article  CAS  Google Scholar 

  27. Ramesh, M. V. and Lonsane, B. K. (1990), Appl. Microbiol. Biotechnol. 33, 501–505.

    Article  CAS  Google Scholar 

  28. Kim, J. H., Hosobuchi, M., Kishmoto, M., Seki, T., and Ryu, D. D. Y. (1985), Biotechnol. Bioeng. 27, 1445–1450.

    Article  CAS  Google Scholar 

  29. Zandrazil, F. and Brunet, H. (1981), Eur. J. Appl. Microbiol. Biotechnol. 11, 183–188.

    Article  Google Scholar 

  30. Lekha, P. K. and Lonsane, B. K. (1994), Process Biochem. 29, 497–503.

    Article  CAS  Google Scholar 

  31. Elad, Y., Chet, I., Boyle, P., and Henis, Y. (1993), Phytopathology 73, 85–88.

    Article  Google Scholar 

  32. Ridout, C. J., Coley-Smith, J. R., and Lynch, J. M. (1988), Enzyme Microb. Technol. 10, 180–187.

    Article  CAS  Google Scholar 

  33. Tokimoto, K. (1982), Trans. Mycol. Soc. Japan 23, 13–20.

    Google Scholar 

  34. Felse, P. A. and Panda, T. (2000), Bioprocess Eng. 23, 127–134.

    CAS  Google Scholar 

  35. Pandey, A. (1995), Ind. J. Microbiol. 35, 335–338.

    Google Scholar 

  36. Berger, L. R. and Reynolds, D. M. (1958), Biochim. Biophys. Acta 29, 522–526.

    Article  CAS  Google Scholar 

  37. Monreal, J. and Reese, E. T. (1969), Can. J. Microbiol. 15, 689–692.

    Article  CAS  Google Scholar 

  38. Shaikh, S. A. and Deshpande, M. V. (1993), World. J. Microbiol. Biotechnol. 9, 468–475.

    Article  CAS  Google Scholar 

  39. Takahashi, M., Tsukiyama, T., and Suzuki, T. (1993), J. Ferment. Bioeng. 75, 457–459.

    Article  CAS  Google Scholar 

  40. Shapira, R., Ordentlich, A., Chet, I., and Oppenheim, A. B. (1989), Phytopathology 79, 1246–1249.

    CAS  Google Scholar 

  41. Jung, W. J., Jung, S. J., An, K. N., Jin, Y. L., Park, R. D., Kim, K. Y., Shon, B. K., and Kim, T. H. (2002), J. Microbiol. Biotechnol. 12, 865–871.

    CAS  Google Scholar 

  42. Lorito, M., Harman, G. E., Hayes, C. K., Broadway, R. M., Tronsmo, A., Woo, S. L., and Di Pietro, A. (1993), Phytopathology 83, 302–307.

    Article  CAS  Google Scholar 

  43. Vyas, P. and Deshpande, M. V. (1989), J. Gen. Microbiol. 35, 343–350.

    CAS  Google Scholar 

  44. Gomes, R. C., Semedo, L. T. A. S., Soares, R. M. A., Linhares, L. F., Ulhoa, C. J., and Coelho, R. R. R. (2001), J. Appl. Microbiol. 90, 653–661.

    Article  CAS  Google Scholar 

  45. Chemin, L., Ismailov, A., Haran, S., and Chet, I. (1995), Appl. Environ. Microbiol. 61, 1720–1726.

    Google Scholar 

  46. Inbar, J. and Chet, I. (1991), Soil Biol. Biochem. 23, 973–978.

    Article  CAS  Google Scholar 

  47. Binod, P., Pusztahelyi, T., Nagy, V., Sandhya, C., Szakács, G., Pócsi, I., and Pandey, A. (2005), Enzyme Microb. Technol. 36, 880–887.

    Article  CAS  Google Scholar 

  48. Rattanakit, N., Yano, S., Wakayama, M., Plikomol, A., and Tachiki, T. (2003), J. Biosci. Bioeng. 95, 391–396.

    CAS  Google Scholar 

  49. Barranco-Florido, J. E., Alatorre-Rosas, R., Gutiérrez-Rojas, M., Viniegra-González, G., and Saucedo-Castañeda, G. (2002), Enzyme Microb. Technol. 30, 910–915.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biotechnology Division, Regional Research Laboratory, CSIR, Trivandrum, 695 019, India

    Chandran Sandhya, Parameswaran Binod, K. Madhavan Nampoothiri & Ashok Pandey

  2. Department of Agricultural Chemical Technology, Technical University of Budapest, Gellert ter 4, Budapes, Hungary

    George Szakacs

Authors
  1. Chandran Sandhya
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Parameswaran Binod
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. Madhavan Nampoothiri
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. George Szakacs
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ashok Pandey
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sandhya, C., Binod, P., Nampoothiri, K.M. et al. Microbial synthesis of chitinase in solid cultures and its potential as a biocontrol agent against phytopathogenic fungus Colletotrichum gloeosporioides . Appl Biochem Biotechnol 127, 1–15 (2005). https://doi.org/10.1385/ABAB:127:1:001

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1385/ABAB:127:1:001

Index Entries

Search

Navigation