Skip to main content

Advertisement

SpringerLink
Log in

Optimization, Purification, and Characterization of Extracellular Mesophilic Alkaline Cellulase from Sponge-Associated Marinobacter sp. MSI032

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

Abstract

Marinobacter sp. (MSI032) isolated from the marine sponge Dendrilla nigra was optimized for the production of extracellular cellulolytic enzyme (CMCase) by submerged fermentation. Initial experiments showed that the culture medium containing 1% maltose as carbon source and 1% peptone and casein as nitrogen source supported maximal enzyme production at 27 °C and at a pH of 9.0. Further optimization carried out showed the maximal enzyme production was supported by the presence of 2% NaCl and 10 mM Zn2+ ions in the production media. The production of enzyme cellulase occurred at 48 h of incubation which proved the importance of this strain for cellulase production in large scale. Further, the enzyme was purified to 12.5-fold with a 37% yield and a specific activity of 2,548.75 U/mg. The purified enzyme displayed maximum activity at mesophilic temperature (27–35 °C) and at a broad pH range with optimal activity at pH 9.0. The purified enzyme was stable even at a higher alkaline pH of 12.0 which is greater than the pH stability that has not been reported in any of the cellulolytic isolates studied so far. Thus, from the present study, it is crucial that, instead of exploring the thermophilic resource that is limited in natural environments, the mesophilic bacteria that occurs commonly in nature can be added up to the database of cellulolytic bacteria. Thus, it is possible that a wide diversity of mesophilic bacteria associated with marine sponges opens up a new doorstep for the degradation of cellulosic waste material for the production of liquid fuels. This is the first report elucidating the prospects of sponge-associated marine bacterium for the production of extracellular alkaline cellulase.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

References

  1. Beguin, P., & Aubert, J. P. (1994). FEMS (Federation of European Microbiological Societies) Microbiology Reviews, 13, 25–58.

    CAS  Google Scholar 

  2. Singh, A., & Hayashi, K. (1995). Advances in Applied Microbiology, 40, 1–44.

    Article  CAS  Google Scholar 

  3. Tomme, P., Warren, R. A., & Gilkes, N. R. (1995). Advances in Microbial Physiology, 37, 1–81.

    Article  CAS  Google Scholar 

  4. Bailey, M. J., & Poutanen, K. (1987). Applied Microbiology and Biotechnology, 30, 5–10.

    Google Scholar 

  5. Bauchop, T., & Mountfort, D. O. (1981). Applied and Environmental Microbiology, 42, 1103–1110.

    CAS  Google Scholar 

  6. Ito, S. (1997). Extremophiles, 1, 61–66.

    Article  Google Scholar 

  7. Uhlig, H. (1998). Industrial enzymes and their applications (p. 435). New York: Wiley. ISBN 0-471-19660-6.

    Google Scholar 

  8. Lee, S. (2005). Encyclopedia of chemical processing. Boca Raton: CRC.

    Book  Google Scholar 

  9. Ito, S., Kobayashi, T., Ara, K., Ozaki, K., Kawai, S., & Hatada, Y. (1998). Extremophiles, 2, 185–190.

    Article  CAS  Google Scholar 

  10. Bajpai, P. (1999). Biotechnology Progress, 15, 147–157.

    Article  CAS  Google Scholar 

  11. Bhat, M. K. (2000). Biotechnology Advances, 18, 355–383.

    Article  CAS  Google Scholar 

  12. Cavaco-Paulo, A. (1998). Carbohydrate Polymers, 37, 273–277.

    Article  CAS  Google Scholar 

  13. Paice, M. G., & Jurasek, L. (1987). Journal of Wood Chemistry and Technology, 4, 187–198.

    Article  Google Scholar 

  14. Poutanen, K. (1997). Trends in Food Science and Technology, 8, 300–306.

    Article  Google Scholar 

  15. Shikata, S., Saeki, K., Okoshi, H., Yoshimatsu, T., Ozaki, K., Kawai, S., et al. (1990). Agricultural and Biological Chemistry, 54, 91–96.

    CAS  Google Scholar 

  16. Bar, D. J. S., Kudo, H., Jakober, K. D., & Cheng, K. J. (1989). Canadian Journal of Botany, 67, 2815–2824.

    Article  Google Scholar 

  17. Christakopoulos, P., Hatzinikolaou, D. G., Fountoukidis, G., Kekos, D., Claeyssens, M., & Macris, B. J. (1999). Archives of Biochemistry and Biophysics, 364, 61–66.

    Article  CAS  Google Scholar 

  18. Coughlan, M. P., Hon-nami, K., Konnami, H. L., Ljungdabl, G., & Paulin, J. J. (1985). Biochemical and Biophysical Research Communications, 130, 904–909.

    Article  CAS  Google Scholar 

  19. Selvin, J., Soniya, J., Asha, K. R. T., Manjusha, W. A., Sangeetha, V. S., Jayaseema, D. M., et al. (2004). Federation of European Microbiological Societies Microbiology, Ecology, 50, 117–122.

    CAS  Google Scholar 

  20. Teather, R. M., & Wood, P. J. (1982). Methods in Enzymology, 160, 59–74.

    Google Scholar 

  21. Collins, C. H., Lyne, P. M., & Grange, J. M. (1989). Microbiological methods. London: Butterworths.

    Google Scholar 

  22. Altschul, S. F., Gise, W., Miller, W., Myers, E. W., & Lipman, D. J. (1990). Journal of Molecular Biology, l215, 403–410.

    Google Scholar 

  23. Altschul, S. F., Thomas, L. M., Alejandro, A. S., Zhang, J., Zhang, Z., Miller, W., et al. (1997). Nucleic Acids Research, 25, 3389–3402.

    Article  CAS  Google Scholar 

  24. Hall, T. A. (1999). Nucleic Acids Symposium Series, 41, 95–98.

    CAS  Google Scholar 

  25. Rani, D. S., & Nand, K. (2000). Process Biochemistry, 36, 355–362.

    Article  CAS  Google Scholar 

  26. Bernfield, P. (1955). Methods in Enzymology, 1, 149–158.

    Article  Google Scholar 

  27. Miller, G. L. (1959). Analytical Chemistry, 37, 426–428.

    Article  Google Scholar 

  28. Lowry, O. H., Rosebrough, N., Farr, A., & Randall, R. (1951). Journal of Biological Chemistry, 193, 265–275.

    CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  30. Coughlan, M. P., & Ljungdahl, L. G. (1988). FEMS symposium no. 43. New York: Academic.

    Google Scholar 

  31. Cavedon, K., Susan, T., Leschine, B., & Canale-parola, E. (1990). Journal of Bacteriology, 172, 4222–4230.

    CAS  Google Scholar 

  32. Saxena, S., Bahadur, J., & Verma, A. (1991). Applied Microbiology and Biotechnology, 34, 668–670.

    Article  CAS  Google Scholar 

  33. Harchand, R. K., & Singh, S. (1997). Acta Microbiologica Immunologica Hungarica, 44, 229–239.

    CAS  Google Scholar 

  34. Ito, S., Shikata, S., Ozaki, K., Kawai, S., Okamoto, K., Inoue, S., et al. (1989). Agricultural and Biological Chemistry, 53, 1275–1281.

    CAS  Google Scholar 

  35. Singh, N. B., & Sobti, R. C. (2001). J. World Journal of Microbiology & Biotechnology, 17, 761–765.

    Article  CAS  Google Scholar 

  36. Fukumori, F., Kudo, T., & Horikoshi, K. (1985). Journal of General Microbiology, 131, 3339–3345.

    CAS  Google Scholar 

  37. Showale, J. G., & Sadana, J. C. (1978). Canadian Journal of Microbiology, 24, 1204–1216.

    Article  Google Scholar 

  38. Song, F. L., Forsberg, C. W., & Gibbins, L. N. (1985). Applied and Environmental Microbiology, 50, 220–228.

    Google Scholar 

  39. Doi, A. H., Park, J. S., Liu, C. C., Malburg, L. M., Tamaru, Y., Ischiishi, A., et al. (1998). Extremophiles, 2, 53–60.

    Article  CAS  Google Scholar 

  40. Lewis, S. M., Montgomery, C. M., Garleb, K. A., Berger, L. L., & Fahey, G. C. (1988). Applied and Environmental Microbiology, 54, 1163–1169.

    CAS  Google Scholar 

  41. Malek, M. A., Chowdhury, N. A., Yousouf, Q. M., & Chaudhury, N. (1988). Enzyme and Microbial Technology, 10, 750–753.

    Article  CAS  Google Scholar 

  42. Pfeffer, J. T. (1974). Biotechnology and Bioengineering, 16, 771–787.

    Article  CAS  Google Scholar 

  43. Hagerdal, B., Harris, H., & Pye, E. K. (1979). Biotechnology and Bioengineering, 21, 345–355.

    Article  CAS  Google Scholar 

  44. Pardo, A. G., & Forchiassin, F. (1998). Revista Argentina de Microbiologia, 30, 20–29.

    CAS  Google Scholar 

  45. Spreinat, A., & Antranikian, G. (1990). Applied Microbiology and Biotechnology, 33, 511–518.

    Article  CAS  Google Scholar 

  46. Hakamada, Y., Endo, K., Takizawa, S., Kobayashi, T., Shirai, T., Yamane, T., et al. (2002). Biochimica et Biophysica Acta, 1570, 174–180.

    CAS  Google Scholar 

  47. Endo, K., Hakamada, Y., Takizawa, S., & Kubota, H. (2001). Applied Microbiology and Biotechnology, 57, 109–116.

    Article  CAS  Google Scholar 

  48. Kim, J. Y., Hur, S. H., & Hong, J. H. (2005). Biotechnology Letters, 27, 313–316.

    Article  CAS  Google Scholar 

  49. Hakamada, Y., Koike, K., Yoshimatsu, T., Mori, H., Kobayashi, T., & Ito, S. (1997). Extremophiles, 1, 151–156.

    Article  CAS  Google Scholar 

  50. Shanmughapriya, S., Krishnaveni, J., Selvin, J., Gandhimathi, R., Arun Kumar, M., Thangavelu, T., et al. (2008). Bioprocess and Biosystems Engineering, 31, 427–433.

    Article  CAS  Google Scholar 

  51. Shanmughapriya, S., Seghal Kiran, G., Selvin, J., Gandhimathi, R., Bastin Baskar, T., Manilal, A., et al. (2008). Biotechnology and Bioprocess Engineering, 14, 67–75.

    Article  CAS  Google Scholar 

  52. Seghal Kiran, G., Shanmughapriya, S., Jayalakshmi, J., Selvin, J., Gandhimathi, R., Sivaramakrishnan, S., et al. (2008). Bioprocess and Biosystems Engineering, 31, 483.

    Article  CAS  Google Scholar 

  53. Riemann, F., & Helmke, E. (2002). Marine Ecology, 23, 93–113.

    Article  CAS  Google Scholar 

  54. Berkenheger, I., & Fischer, U. (2004). International Microbiology, 7, 1139–6709.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Microbiology, Bharathidasan University, Tiruchirappalli, 620 024, India

    S. Shanmughapriya, Joseph Selvin, T. Anto Thomas & C. Rani

  2. Department of Biotechnology, Bharathidasan University, Tiruchirappalli, 620 024, India

    G. Seghal Kiran

Authors
  1. S. Shanmughapriya
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. Seghal Kiran
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Joseph Selvin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T. Anto Thomas
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C. Rani
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Joseph Selvin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shanmughapriya, S., Kiran, G.S., Selvin, J. et al. Optimization, Purification, and Characterization of Extracellular Mesophilic Alkaline Cellulase from Sponge-Associated Marinobacter sp. MSI032. Appl Biochem Biotechnol 162, 625–640 (2010). https://doi.org/10.1007/s12010-009-8747-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-009-8747-0

Keywords

Search

Navigation