Screening and Isolation of Collagenase Producing Microorganism from Proteins Waste Found in Himalayan Region

Document Type: Original Article

Authors

Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla, India

Abstract

Multicellular organisms of the human and animal bodies such as skin, bones, tendons, cartilage and blood vessels have collagen as a fibrous element. Collagen is a most representative protein of the leather wastes and collagenase is a proteolytic enzyme. The collagenase is responsible for the degradation of native collagen to small peptide fragments, without affecting the other proteins. Collagenase has widespread applications such as cosmetics, wound healing, diabetic ulcer, arterial ulcers and burns surgery, etc. In the present study, Twenty-seven collagenase-producing microorganisms were identified and isolated from soil/sewage samples of the fish market and slaughterhouse areas. CS-20 sample was found most efficient producer microorganism of collagenase. This sample was identified from Pseudomonas genus and isolated as Pseudomonas species. The parameters such as temperature, substrate, pH, incubation period and inoculum percentage were also optimized to screen the hyper producer strain with maximum collagenase activity. CS-20 isolate (Collagenase producing microorganism) was capable of hydrolyzing other protein substrates such as gelatine and azocoll. Thus, CS-20 isolate is high-efficiency strain for the production of collagenase.

Keywords


  1. Anbu, P., Gopinath, S., Cihan, A.C., Chaulagain, B.P., Microbial enzymes and their applications in industries and medicine. Biomed Res Int, 2013, Vol. 2013, pp. 1-2.
  2. Chauhan, B., Gupta, R., Application of statistical experimental design for optimization of alkaline protease production from Bacillus sp. RGR-14. Process Biochem, 2004, Vol. 39, pp. 2115-2122.
  3. Adrio, J.L., Demain, A.L., Microbial Enzymes: Tools for biotechnological processes. Biomol, 2014, Vol. 4, pp. 117-139.
  4. Gautam, M., Azmi, W., Purification of extracellular collagenase from Pseudomonas sp: remarkable collagenolytic activity. Adv Biotech Microbiol, 2017, Vol. 4(2), pp. 1-8.
  5. Lima, C.A., Rodrigues, P.M.B., Porto, T.S., Viana, D.A., Lima Filho, J.S., Porto, A.L.F., Carneiro-daCunha, MG., Production of a collagenase from Candida albicans URM3622. Biochem Eng J, 2009, Vol. 43, pp. 315-320.
  6. Goldberg, G.I., Wilhelm, S.M., Kronberger, A., Bauer, E.A., Grant, G.A., Eisen, A.Z., Human fibroblast collagenase: complete primary structure and homology to an oncogene transformation-induced rat protein. J Biol Chem, 1986, Vol. 261, pp. 6600-6605.
  7. Park, P.J., Lee, S.H., Byun, H.G., Kim, S.H., Kim, S.K., Purification and characterization of a collagenase from the Mackerel, Scomber japonicas. J Biochem Mol Biol, 2002, Vol. 35, pp. 576-582.
  8. Kim, S.K., Park, P.J., Kim, J.B., Shahidi, F., Purification and characterization of a collagenolytic protease from the filefish, Novoden modestrus. J Biochem Mol Biol, 2002, Vol. 335, pp. 165-171.
  9. Schenck, S., Theodore Chase, J.R., Rosenzweig, W.D., Pramer, D., Collagenase production by nematode-trapping fungii. Appl Environ Microbiol, 1980, Vol. 40, pp. 567-570.
  10. Anandan, D., Marmer, W.N., Dudley, R.L., Isolation, characterization and optimization of culture parameters for production of an alkaline protease isolated from Aspergillus tamarii. J Ind Microbiol Biotechnol, 2007, Vol. 34, pp. 339-347.
  11. Beltran, A., Bonnet, M., Quali, A., Collagenase effect on thermal denaturation of intramuscular collagen. J Food Sci, 2006, Vol. 56, pp. 1497-1499.
  12. Gutierrez-Fernandez, A., Inada, M., Balbin, M., Increased inflammation delays wound healing in mice deficient in collagenase-2 (MMP-8). FASEB J, 2007, Vol. 21, pp. 2580-2591.
  13. Kanth, S.V., Venba, R., Madhan, B., Chandrababu, N.K. and Sadulla, S., Studies on the influence of bacterial collagenase in leather dyeing. Dyes Pigm, 2008, Vol. 76, pp. 338-347.
  14. Brand, J.S., Hefley, T.J., Collagenase and the isolation of cells from bone. In: Pretlow, T.G. (ed.): Cell separation methods and selected applications. Acad Press, New York, 1984, Vol. 3, pp. 265-283.
  15. Patel, V.A., Logan, A., Watkinson, J.C., Uz-Zaman, S., Sheppard, M.C., Ramsden, J.D., Eggo, M.C., Isolation and characterization of human thyroid endothelial cells. Am J Physiol Endocrinol Metab, 2003, Vol. 284, pp. 168-176.
  16. Marcus, G.J., Connor, L., Domingo, M.T., Tsang, B.K., Downey, B.R., Ainsworth, L., Enzymatic dissociation of ovarian and uterine tissues. Endocr Res, 1984, Vol. 10, pp. 151-62.
  17. Ray, J.L., Leach, R., Herbert, J.M., Benson, M., Isolation of vascular smooth muscle cells from a single murine aorta. Methods Cell Sci., 2001, Vol. 23, pp. 185-188.
  18. Ravanti, L., Kahari, V.M., Matrix metalloproteinases in wound repair. Int J Mol Med, 2000, Vol. 6, pp. 391.
  19. Anwar, A., Saleemuddin, M., Alkaline-pH acting digestive enzymes of the polyphagous insect pest Spilosoma obliqua: stability and potential as detergent additives. Biotechnol Appl Biochem, 1997, Vol. 25(1), pp. 43-46.
  20. Gousterova, A., Lilova, A., Trenev, M., Spassov, G., Nedkov, P., Thermophilic actinomycetes as producers of Collagenase. Compt Rend Acad Bulg Sci, 1998, Vol. 51, pp. 71-76.
  21. Wu, C.L., Li, C., Chen, H., Shuliang, L., Purification and characterization of a novel collagenase from Bacillus pumilus Col-J. Appl Biochem Biotechnol, 2010, Vol. 160, pp.129-134.
  22. Liu, L., Ma, M., Cai, Z., Yang, X., Wang, W., Purification properties of a collagenolytic protease produced by Bacillus cereus MBL13 Strain. Food Technol Biotechnol, 2010, Vol. 48, pp. 151-160.
  23. Nagano, H., Kim, A., Purification of collagenase and specificity of its related enzyme from Bacillus subtilis FS-2. Biosci Biotechnol Biochem, 1999, Vol. 63, pp. 181-183.
  24. Kameyama, S., Akama., Titration of kappa toxin of Clostridium perfringens and kappa antitoxin by a modified azocoll method. Jpn J Med Sci Biol, 1970, Vol. 23, pp. 31-46.
  25. Finland, M., Jones, W.F., Barnes, M.W., Occurrence of serious bacterial infections since the introduction of antibacterial agents. J Am Med Assoc, 1959, Vol. 170, pp. 2188-2197.
  26. Iococca, V.F., Sibinga, M.S., Barbero, G.J., Respiratory tract bacteriology in cystic fibrosis. Amer J Dis Child, 1963, Vol. 106, pp. 315-324.
  27. Endo, A., Murakawa, S., Shimizu, H., Shiraishi, Y., Purification and properties of collegenase from a Streptomyces Sp. J Bacteriol, 1986, Vol. 102, pp. 163-170.
  28. Merkel, J.R., Dreisbach, J.H., Purification and characterization of a marine bacterial collagenase. Biochem J, 1978, Vol. 17, pp. 2857-2863.
  29. Reid, G.C., Robb, F.T., Woods, D.R., Regulation of extracellular collagenase production in Achromobacter iophagus. J Gen Microbiol, 1978, Vol. 109, pp. 149-154.
  30. Mandl, I., Maclennan, J.D., Howes, E.L., Isolation and characterization of proteinase and collagenase from Clostridium histolyticum. J Clin Invest, 1953, Vol. 32, pp. 1323-1329.
  31. Ok, T., Hashinaga, F., Detection and Production of Extracelluar collagenolytic Enzyme from Zygosaccharomyces rouxii. J Gen App Microbiol, 1996, Vol. 42, pp. 517-523.
  32. Okamoto, M., Yonejima, Y., Tsujimoto, Y., Suzuki, Y., Watanabe, K., A thermostable collagenolytic protease with a very large molecular mass produced by thermophilic Bacillus sp. strain MO-1. Appl Microbiol Biotechnol, 2001, Vol. 57, pp. 103-108.
  33. Jain, R., Jain, P.C., Production and partial characterization of collagenase of. Streptomyces exfoliates CFS 1068 using poultry feather. Ind J Exp Biol, 2010, Vol. 48, pp. 174-178.
  34. Mukherjee, J., Webster, N., Llewellyn, L., Purification and characterization of a collagenolytic enzyme from a pathogen of the great barrier reef sponge, Rhopaloeides odorabile. PLoS One, 2009, Vol. 4, pp. e7177.
  35. Baehaki, A., Suhartono, M.T., Sukarno, S.D., Sitanggang, A.B., Setyahadi, S., Meinhardt, F., Purification and characterization of collagenase from Bacillus licheniformis F11.4.  Afr J Microbiol Res, 2012, Vol. 6, pp. 2373-2379.
  36. Hamdy, H.S., Extracellular collagenase from Rhizoctonia solani: Production, purification and characterization. Iran J Biotechnol, 2008, Vol. 7, pp. 333-340.
  37. Brinckerhoff, C.E., Rutter. J.L., Benbow, U., Interstitial collagenases as markers of tumor progression. Clin Cancer Res, 2000, Vol. 6, pp. 4823-4830.
  38. Matasushita, O., Koide, T., Kobayashi, R., Nagata, K., Okabe, A., Substrate recognition by collagen binding domain of Clostridium histolyticum class I collagenase. J Biol Chem, 2001, Vol. 276, pp. 8761-8770.
  39. Jin, B., Alter, H.J., Zhang, Z.C., Shih, J.W., Esteban, J.M., Sun, T., Yang, Y.S.,  Qiu, Q., Liu, X.L.,  Yao, L., Wang, H.W., Cheng, L.F., Reversibility of experimental rabbit liver cirrhosis by portal collagenase administration. Labo Invest, 2005, Vol. 85, pp. 992-1002.