Isolation, Screening and Production of Bacterial Protease Enzyme

Authors

  • Rajee T  Department of Biotechnology, Muthayammal College of Arts and Science, Rasipuram,Tamil Nadu, India
  • Selvamaleeswaran P  Department of Biotechnology, Muthayammal College of Arts and Science, Rasipuram,Tamil Nadu, India

Keywords:

Protease, Poultry Waste Soil, Skim Milk Agar, Zone Of Hydrolysis, Fermentation, Staphylococcus.

Abstract

The objective of present study was to isolate and screen proteolytic bacteria from poultry waste site at Rasipuram, Namakkal district, Tamilnadu, India. Soil samples were serially diluted and 0.1ml of sample was spreaded on nutrient agar plate at 37o C for 48 hrs. Four bacterial colonies isolated were screened for proteolytic activity by skim milk agar plate assay. The isolate RPB 2 showed maximum zone of hydrolysis (13mm) was selected and identified as Staphylococcus sp by morphological and biochemical test. Various factors influencing the protease production such as incubation time, initial pH, incubation temperature, inoculum level and casein concentration were studied. The results showed that maximum protease enzyme production occurred at incubation time 48hr (34 U/ml), pH 7.0 (42U/ml), 2% inoculum concentration (21.5U/ml), temperature 37o C (51U /ml) and 2% casein concentration (72U/ml).

References

  1. Sari, E., Lo?o?lu, E. and Oktemer, A. (2015). Purification and characterization of organic solvent stable serine alkaline protease from newly isolated Bacillus circulans M34. Biomedical Chromatography, 29(9): 1356-1363.
  2. Rupali, D. (2015). Screening and Isolation of Protease Producing Bacteria from Soil Collected from Different Areas of Burhanpur Region (MP) India. International Journal of Current Microbiology and Applied Sciences, 4(8): 597-606.
  3. Safia, B. M. Umar, D. and Naqvi,S.H.A. 2016. Optimization of culture conditions for the production of protease by Pleurotus eryngii, Pak. J.Biotechnol, 13(3): 193- 198.
  4. Josephine, F. S., Ramya, V. S., Devi, N., Ganapa, S. B. and Vishwanatha, T. (2012). Isolation, production and characterization of protease from Bacillus sp. isolated from soil sample. Journal of Microbiology and Biotechnology Research, 2(1): 163-168.
  5. Zhang, W. W., Sun, K., Cheng, S. and Sun, L. (2008). Characterization of DegQVh, a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain. Applied and environmental microbiology, 74(20): 6254-6262.
  6. Butterworth, M. B., Zhang, L., Heidrich, E. M., Myerburg, M. M. and Thibodeau, P. H. (2012). Activation of the epithelial sodium channel (ENaC) by the alkaline protease from Pseudomonas aeruginosa. Journal of Biological Chemistry, 287(39): 32556-32565.
  7. Munoz-Tamayo, R., De Groot, J., Bakx, E., Wierenga, P. A., Gruppen, H., Zwietering, M. H. and Sijtsma, L. (2011). Hydrolysis of ?-casein by the cell-envelope-located P I-type protease of Lactococcus lactis: A modelling approach. International Dairy Journal, 21(10): 755-762.
  8. Marti, M., Trotonda, M. P., Tormo-Mas, M. Á., Vergara-Irigaray, M., Cheung, A. L., Lasa, I. and Penadés, J. R. (2010). Extracellular proteases inhibit protein-dependent biofilm formation in Staphylococcus aureus. Microbes and infection, 12(1): 55-64.
  9. Abirami, V., Meenakshi, S.A., Kanthymathy, K., Bharathidasan, R., Mahalingam, R. and Panneerselvam, A. (2011). Partial Purification and Characterization of an Extracellular Protease from Penicillium janthinellum and Neurospora crassa. Eur. J. Exp. Biol, 1(3): 114-123.
  10. Palsaniya, P., Mishra, R., Beejawat, N., Sethi, S. and Gupta, B.L. (2012). Optimization of alkaline protease production from Bacteria isolated from soil. J. Microbiol. Biotech. Res, 2(6): 858-865.
  11. Sony, I. S. andPotty, V. P. (2016). Isolation and Identification of Protease Producing Bacteria from Food Processing Industries. Int. J. Curr. Microbiol. App. Sci, 5(3): 181-189.
  12. Holt, J.G., Krieg, N.R., Sneath, P.H.A., Staley, J.T and Williams, S.T.(1994). Bergeys Manual of Determinative Bacteriology, 9th edn. Baltimore: Williams &Wilkins.
  13. Lakshmi, B. K. M., Sri, P. R., Devi, K. A. and Hemalatha, K. P. J. (2014). Media optimization of protease production by Bacillus licheniformis and partial characterization of alkaline protease. Int. J. Curr. Microbiol, App. Sci. 3(5): 650-659.
  14. Ire, F. S., Okolo, B. N., Moneke, A. N. and Odibo, F. J. (2011). Influence of cultivation conditions on the production of a protease from Aspergillus carbonarius using submerged fermentation. African Journal of Food Science, 5(6): 353-365.
  15. Ellaiah P, Adinarayana K, Bhavani Y, Padmaja P. and Srinivasulu B (2002). Optimization of process parameters for glucoamylase production under solid-state fermentation by a newly isolated Aspergillus sp., Process Biochem,38 (4): 615-620.
  16. Tiwari, O. N., Devi, T. B., Devi, K. S., Oinam, G., Indrama, T., Ojit, K. and Ningshen, L. (2015). Isolation and optimization of alkaline protease producing Bacteria from undisturbed soil of NE-region of India falling under Indo-Burma biodiversity hotspots. Journal of Applied Biology & Biotechnology, 3(4): 025-031.
  17. Radha,S., Hari Prasad,T., Syamala Devi, N. and Mamata,S. (2014). Fed batch and batch submerged fermentation for alkaline protease production from mixed consortium of Pseudomonas putida and Staphylococcus aureus. Advances in Applied Science Research, 5(3):31 5-324.
  18. Gençkal, H. and Tari, C. (2006). Alkaline protease production from alkalophilic Bacillus sp. isolated from natural habitats. Enzyme and Microbial Technology, 39(4): 703-710.
  19. Niyonzima, F. N.and More, S. S. (2013). Screening and optimization of cultural parameters for an alkaline protease production by Aspergillus terreus under submerged fermentation. Int J Pharm Bio Sci, 4(1): 1016-1028.
  20. Singh, S. and Das, S. (2011). Screening, production, optimization and characterization of cyanobacterial polysaccharide. World J. Microbiol. Biotechnol, 27: 1971- 1980.
  21. Li, P., Harding, S.E. and Liu, Z. (2001). Cyanobacterial Exopolysaccharides: their nature and potential biotechnological applications. Biotechnol. Genet. Eng, 18: 375 404.
  22. Bajaj, I.B., LeLe, S.S. and Singhal, R.S. (2009). A statistical approach to optimization of fermentive production of poly (gamma glutamic acid) from Bacillus licheniformis NCIM 2324. Bioresour. Technol, 100: 826- 832.
  23. Anustrup, K. (1980). Proteinase. In: Rose, A.H. (Ed.), Economic microbiology. Microbial enzymes and bioconversions. Academic Press, New York. Pp. 50 114.
  24. Sabu, A., Sarita, S., Pandey, A., Boga,r B., Szakacs, G. and Socol, C.R.(2012). Solid-state fermentation for production of phytase by Rhizopus oligosporus. Appl Biochem Biotechnol, 103, 251-260.
  25. Abusham, R A., Rahman, R. J., Salleh, A. B. and Basri, M. (2009). Optimization of physical factors affecting the production of thermostable organic solvent tolerant protease from a newly isolated halo tolerant Bacillus subtilis strain R and. Microbial Cell Factories, 8(20):1475-2859.
  26. Jenitta, X. J., Priya, S. E.and Gnanadoss, J. J. (2015). Optimization of culture conditions and inducers for improved protease production by Penicillium griseofulvum LCJ231 under submerged fermentation. International Journal of Advanced Biotechnology and Research, 6(2): 152-60.
  27. Patil, P., Sabale, S.and Devale, A. (2015). Isolation and characterization of protease producing bacteria from rhizosphere soil and optimization of protease production parameters. International Journal of Current Microbiology and Applied Sciences, 2: 58-64.

International Journal of Scientific Research in Science, Engineering and Technology

Downloads

Published

2017-06-30

Issue

Volume 3, Issue 3, May-June-2017

Section

Research Articles

License

Copyright (c) IJSRSET

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

How to Cite

[1]
Rajee T, Selvamaleeswaran P, " Isolation, Screening and Production of Bacterial Protease Enzyme, International Journal of Scientific Research in Science, Engineering and Technology(IJSRSET), Print ISSN : 2395-1990, Online ISSN : 2394-4099, Volume 3, Issue 3, pp.447-452, May-June-2017.