Production and Characterization of Collagenase From Bacillus sp. 6-2 Isolated From Fish Liquid Waste

Article History

Submited : January 14, 2019
Published : May 30, 2019

Collagenases are enzyme that are able to hydrolyze native collagen into fragment collagen peptides. Collagenases and its hydrolysis products have received tremendous attention in medical and industrial applications. The present study was conducted to isolate and identify new collagenase producing bacteria from fish liquid waste, then produce and characterize collagenase. A total of 7 isolate from fish liquid waste were screened on selective medium containg 2 % collagen and its activity was confirmed by the formation of clear zone. Isolat 6-2 was positif as collagenase producer and identified as Bacillus sp. 6-2 by morphological and biochemical characteristics. The optimum fermentation time of enzyme was investigated. Collagenase crude extract was characterized by the effect of pH, temperature, and metal ions. Isolat 62 optimally produced collagenase enzyme after 30 h of incubation with activity of   0.072 U/mL and protein content of 3.768 mg/mL. The optimum pH and temperature were 7.0 and 40 oC, respectively. The enzyme was activated by 1 mM Ca2+and  Mg2+, and inhibited by   1 mM  Zn2+ and Co2+. Collagenase from Bacillus sp. 6-2 may have potentials for medical and industrial applications.

References

  1. Alipour, H., Raz, A., Zakeri, S., and Djadid, N.D. 2016. Therapeutic Applications of Collagenase (Metalloproteases): A Review. Asian Pac J Trop Biomed; 6 (11); 975–981.
  2. Baehaki, A., Suhartono, M.T., Sukarno, Syah, D., Sitanggang, A.B., Setyahadi, S., and Meinhardt, F. 2012. Purification and Characterization of Collagenase from Bacillus licheniformis F11.4. AJMR; 6 (10); 2373-2379.
  3. Bergmeyer, H.U. 1983. Methods of Enzymatic Analysis, Vol 2. Germany: Verlag Chemie. 1983.
  4. Bhunia, B., Basak, B., Mandal, T., Bhattacharya, P., and Dey, A. 2013. Effect of pH and Temperature on Stability and Kinetics of Novel Extracellular Serine Alkaline Protease (70 kDa). Int. J. Biol. Macromol; 54; 1-8.
  5. Bousopha, S., Nalinanon, S., and Sriket, C. 2016. Production of Collagen Hydrolysate with Antioxidant Activity from Pharaoh Cuttlefish Skin. CMU J. Nat. Sci; 15 (2); 151-162.
  6. Chauhan, A., and Prabha, V. 2017. Production and Partial Purification of Collagenase from Bacillus sp. Isolated from Soil Sample. Int J Adv Res; 2 (5); 60-65.
  7. Gautam, M., and Azmi, W. 2017. Screening and Isolation of Collagenase Producing Microorganism from Proteins Waste Found in Himalayan Region. JABR; 4 (1); 558-565.
  8. Hashim, P., Ridzwan, M.M.S., Bakar, J., and Hashim, D.M. 2015. Collagen in Food and Beverage Industries. IFRJ; 22 (1); 1-8.
  9. Howes, J.M., Pugh, N., Knäuper, V.,and Farndale R.W. 2015. Modified platelet deposition on matrix metalloproteinase 13 digested collagen I. J Thromb Haemost; 13(12); 2253-2259.
  10. Kang, S.I., Jang, Y.B., Choi, Y.J., and Kong, J.Y. 2005. Purification and Properties of a Collagenolytic Produced by Marine Bacterium Vibrio vulnificus CYK279H. Biotechnol. Bioprocess Eng; 10; 593-598.
  11. Liu, L., Ma, M., Cai, Z., Yang, X., and Wang, W. 2010. Purification and Properties of a Collagenolytic Protease Produced by Bacillus cereus MBL13 Strain. Food Technol. Biotechnol; 48 (2); 151–160.
  12. Lowry, O.H., Rosebrough, N.J., Farr, A.I., and Randall. 1951. Protein Measurement with the Folin Phenol Reagen. J.Biol.Chem; 193; 265-275.
  13. Mohammad, A. W. 2014. Process for Production of Hydrolysed Collagen from Agriculture Resources: Potential for Further Development. J. Applied Sci; 14 (12); 1319-1323.
  14. Natsir, H., Patong, R., Suhartono, M.T., and Ahmad, A. 2013. Isolation and Purification of Thermostable Chitinase Bacillus licheniformis Strain HSA3-1a from Sulili Hot Spring in South Sulawesi, Indonesia. Int J Pharm Bio Sci; 4 (3); 1252-1259.
  15. Savita, K., and Arachana, P. 2015. Production of Collagenase by Bacillus KM369985 Isolated from Leather Sample. Int. J. Res. Biosciences; 4 (4); 81-87.
  16. Song, H., and Li, B. 2017. Beneficial Effects of Collagen Hydrolysate: A Review on Recent Developments. Biomed J Sci & Tech Res; 1 (2); 1-4.
  17. Wanderleya, M.C.D.A., Neto, J.M.W.D., Albuquerqueb, W.W.C., Marques, D.D.A.V., Lima, C.D.A., Silverio, S.I.D.C., Filho, J.L.D.L.,
  18. Teixeira, J.A.C., and Porto, A.L.F. 2017. Purification and Characterization of A Collagenase from Penicillium sp. UCP 1286 by Polyethylene Glycol-Phosphate Aqueous Two-Phase System. Protein Expr. Purif; 133; 8-14.
  19. Wen-Jia, P., Jun-Wei, Y., Ya-Nan W, et al. 2012. Matrix metalloproteinases: a review of their structure and role in systemic sclerosis. J Clin Immunol.; 32(6); 1-4.
  20. Wu, Q., Li, C., Li, C., Chen, H., and Shuliang, L. 2010. Purification and Characterization of a Novel Collagenase from Bacillus pumilus Col-J. Appl Biochem Biotechnol; 160; 129–139.
  21. Zhang, Z., Li, G., and Shi, B. 2005. Physicochemical properties of collagen, gelatin and collagen hydrolysate derived from bovine limed split wastes. J.Soc.Leather Technol. Chem; 90; 23-28.

Downloads

Download data is not yet available.
Fulltext