Main Article Content

Abstract

This study aimed to isolate and identify the Bacillus licheniformis bacteria that produces the enzyme collagenase. Due to the selective nature of this enzyme in breaking down and degrading collagen, it was used to tenderize aged buffalo meat. The bacterial isolates were screened using a selective medium and then grown on collagen-agar medium to identify enzyme-producing isolates. Thirteen enzyme-producing isolates were obtained based on the formation of a transparent corona around the colony. Biochemical assessments of the bacterial isolates were carried out using the ABIS online program. The isolates BL1–BL9, BL12 and BL13 were found to be related to B. licheniformis with a percentage match of 98%, while BL10 and BL11 were related to Bacillus pumilus and Bacillus subtilis with a percentage match of 95% and 95.3%, respectively. The enzyme activity of the isolates varied, with the highest activity reaching 200.71 units.ml-1 for isolate BL12. The enzyme of this isolate was chosen to complete the other tests. The optimal time required to obtain the highest enzymatic activity was found to be 20 h, which amounted to 210.25 units.ml-1, with a specific activity of 135.64 units.ml-1. The enzyme was partially purified using ammonium sulphate at a saturation rate of 20–70% as it showed an enzymatic activity and specificity of 355.81 units.ml-1 and a specific activity of 423.58 units.ml-1. Partially purified collagenase was used to tenderize aged buffalo meat. The results showed that treating aged buffalo meat with the enzyme solution for different immersion durations improved the pH, water-holding capacity (WHC) and cooking yield, with an increase in the concentration of hydroxyproline and a significant percentage of soluble collagen; the treated samples showed the best properties with a 60-min immersion duration.

Keywords

Bacillus Biochemical test Buffalo meat Collagenase Tenderization

Article Details

How to Cite
Al-Temimi, W. K. A. ., Aziz, S. N. ., & Khalaf, A. A. . (2023). Production of Partially Purified Collagenase from Bacillus licheniformis and It’s Use to Tenderize Aged Buffalo Meat. Basrah Journal of Agricultural Sciences, 36(1), 75–89. https://doi.org/10.37077/25200860.2023.36.1.07

References

  1. Abdeldaim, M. H., & Ali, H. G. M. (2013). Tenderization of camel meat by using fresh ginger (Zingiber officinale) extract. Food Science and Quality Management, 21, 12-25.
  2. https://iiste.org/Journals/index.php/FSQM/article/view/10470
  3. Adinarayana, K., Ellaiah, P., & Prasad, D. S. (2003). Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11. An Official Journal of the American Association of Pharmaceutical Scientists, 4(4), E56.
  4. https://link.springer.com/article/10.1208/pt040456
  5. Al-Abadi, H. H. M., & Al-Temimi, W. K. A. (2022). Production of hydrolyzed collagen from common carp Cyprinus carpio L. scales by synergistic enzyme systems. Basrah Journal of Agricultural Sciences, 35(1), 94-105.
  6. https://doi.org/10.37077/25200860.2022.35.1.08
  7. Al-Temimi, W. K. A. (2014). Purification and Characterization of Collagenase Extracted from Fish Stomach by Products and Study it’s Tenderizing Meat. Ph. D. Thesis, Univ. Basrah: 151pp. (In Arabic, English abstract).
  8. Anandh, M. A. (2013). Effect of Different Tenderizers on Tenderness and Quality of Buffalo Tripe. Asian Journal. Dairy and Food Research, 32(2), 144-148.
  9. https://arccjournals.com/journal/asian-journal-of-dairy-and-food-research/ARCC347
  10. Antonio, B. (2005). Buffalo Production and Research. FAO. 00100 Rome, 316pp.
  11. AOAC. (2016). Official methods of anaylsis. Retrieved from http://www.eoma.aoac.org/metho ds/info.asp?ID=16264
  12. Badr, H. M. (2008). Tenderness properties and microbial safety of spent hen meat treated by papain and gamma irradiation. Journal of Radiation Research and Applied Sciences, 1(2), 443-462.
  13. https://www.sciencedirect.com/journal/journal-of-radiation-research-and-applied-sciences
  14. Banoon, S. R., & Ali, Z. M. (2018). Identification the local thermophilic strains of Bacillus licheniformis by VITEK-2 system and Gyrase B gene. Al-Kufa University Journal for Biology, 10(2), 157-167.
  15. https://journal.uokufa.edu.iq/index.php/ajb/article/view/8125
  16. Bassetto, F., Maschio, N., Abatangelo, G., Zavan, B., Scarpa, C., & Vindigni, V. ( 2016). Collagenase from Vibrio alginolyticus cultures: Experimental study and clinical perspectives. Annals of Surgical Innovation and Research, 23(6), 557-562.
  17. https://doi.org/10.1177/1553350616660630
  18. Bergeyer, H. U. (1974). Methods of Enzymatic Analysis, Vol. 2, Second Edition, Academic Press, Elsevier Inc., 682pp.
  19. https://www.sciencedirect.com/book/9780120913022/methods-of-enzymatic-analysis
  20. Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254.
  21. https://doi.org/10.1016/0003-2697(76)90527-3
  22. Dumitru, M., Vodnar, D. C., Elemer, S., Ciurescu, G., Habeanu, M., Sorescu, I., Georgescu, S. E., & Dudu, A. (2021). Evaluation of non-encapsulated and microencapsulated lactic acid bacteria. Applied Science, 11(9867), 1-15.
  23. https://doi.org/10.3390/app11219867
  24. Feng, Y. H., Zhang, S. S, Sun, B. Z., Wen, P. X. K. X., & Xu, C. C. (2020). Changes in physical meat traits, protein solubility, and the microstructure of different beef muscles during post-mortem aging. Foods, 806(9), 1-10.
  25. https://doi.org/10.3390/foods9060806
  26. Gecgel, U., Yilmaz, I., Soysal, M. I., Gurcan, E. K., & Kok, S. (2019). Investigating proximate composition and fatty acid profile of Longissimus dorsi from Anatolian water Buffaloes (Bubalus bubalis) raised in similar conditions. Food Science and Technology, 39(4), 830-836.
  27. https://doi.org/10.1590/fst.08918
  28. Geles, K., Poschl, E., & Aiagner, T. (2003). Collagens-structure, function, and biosynthesis. Advanced Drug Delivery Reviews, 28, 1531-1546.
  29. https://doi.org/10.1016/j.addr.2003.08.002
  30. Habtu, E., Mekonnen, B., Kiros, H., Fesseha, H., & Getachew, B. (2020). Meat tenderization of efficiency of papain, bromelain and Zingiber officinale on old aged beef carcass of local zebu cattle. Journal of Trends in Technical & Scientfic Research, 4(1), 9-15.
  31. https://doi.org/10.19080/TTSR.2020.04.555628
  32. Irurueta, M., Cadoppi, A., Langman, L., Grigioni, G., & Carduza, F. (2008). Effect of aging on meat characteristics from water buffalo grown in delta del Paraná in Argentina. Meat Science, 79(3), 529-533.
  33. https://doi.org/10.1016/j.meatsci.2007.12.010
  34. Jin, P., Zhang, L., Yuan, P., Kang, Z., Du, G., & Chen, J. (2016). Efficient biosynthesis of polysaccharides chondroitin and heparosan by metabolically engineered Bacillus sublitis. Carbohydrate Polymers, 140, 424-432.
  35. https://doi.org/10.1016/j.carbpol.2015.12.065
  36. Kandeepan, G., Anjaneyulu, A. S. R., Kondaiah, N., Mendiratta, S. K., & Lakshmanan, V. (2009). Effect of age and gender on the processing characteristics of buffalo meat. Meat Science, 83, 10-14.
  37. https://doi.org/10.1016/j.meatsci.2009.03.003
  38. Liu, L., Ma, M., Cai, Z., Yang, X., & Wang, W. (2010). Purification and properties of a collagenolytic protease produced by Bacillus cereus MBL13 strain. Food Technology and Biotechnology, 48(2), 151-160.
  39. https://hrcak.srce.hr/53624
  40. Moon, S. S. (2018). Effect of proteolytic enzymes and ginger extract on tenderization of M. pectoralis profundus from Holstein steer. Korean Journal for Food Science of Animal Resources, 38(1), 143-151.
  41. https://doi.org/10.5851/kosfa.2018.38.1.143
  42. Naveena, B. M., Mendiratta, S. K., & Anjaneyulu, A. S. R. (2004). Tenderization of buffalo meat using plant proteases from Cucumis trigonus roxb (Kachri) and Zingiber officinale roscoe (Ginger rhizome). Meat Science, 68, 363-369.
  43. https://doi.org/10.1016/j.meatsci.2004.04.004
  44. Naveena, B. M., Kiran, M., Sudhakar, K. R., Ramkrishna, C., Vaithiyanathan, S., & Devetkal, S. K. (2011). Effect of ammonium hydroxide on ultrastructure and tenderness of buffalo meat. Meat Science, 88, 727-732.
  45. https://doi.org/10.1016/j.meatsci.2011.03.005
  46. Parte, A. C. (2018). LPSN-list of prokaryotic names with standing in nomenclature (bactrio.net), 20 years on. International Journal of Systematic and Evolutionary Microbiology, 68(6), 1825-1829. https://10.1099/ijsem.0.002786
  47. Quaglia, G. B., & Gennaro, L. (2003). Enzymes: Uses in Food Processing. Pp., 2125-2139. In Encyclopedia of Food Sciences and Nutrition. Second Edition.
  48. https://doi.org/10.1016/B0-12-227055-X/00420-X
  49. Sartika, D., Natsir, H., Dali, S., & Leliani, L. (2019). Production and characterization of collagenase from Bacillus sp. 6-2 isolated from fish liquid waste. Jurnal Akta Kimia Indonesia, 2(1), 58-66.
  50. https://doi.org/10.20956/ica.v12i1.5924
  51. Savita, K., & Arachana, P. (2015). Production of collagenase by Bacillus KM369985 isolated from leather sample. International Journal of Research in Biosciences, 4(4), 81-87.
  52. http://www.ijrbs.in/index.php/ijrbs/article/view/177
  53. Schroeter, R., Hoffmann, T., Voigt, B., Meyer, H., Bleisteiner, M., Muntel, J., Jurgen, B., Albrecht, D., Becher, D., Lalk, M., Evers, S., Bongaerts, J., Maurer, K. H., Putzer, H., Hecker, M., Schweder, T., & Bremer, E. (2013). Stress responses of the industrial workhorse Bacillus licheniformis to osmotic challenges. PLOS One, 8(11), 1-22.
  54. https://doi.org/10.1371/journal.pone.0080956
  55. Singh, P. K., Ahlawat, S. S., Sharma, D. P., & Pathera, A. (2018). Carcass characteristics of male buffalo calf & meat quality of its veal. Buffalo Bulletin, 37(2), 129-144.
  56. https://kuojs.lib.ku.ac.th/index.php/BufBu/article/view/134
  57. Sorushanova, A., Delgado, L. M., Wu, Z., Shologu, N., Kshirsagar, A., Raghunath, R., Mullen, A. M., Bayon, Y., Pandit, A., Raghunath, M., & Zeugolis, D. I. (2018). The collagen suprafamily: From biosynthesis to advanced biomaterial development. Advanced Materials, 31(1), 1801651-1801690.
  58. https://doi.org/10.1002/adma.201801651
  59. Stoica, C., & Sorescu, I. (2017). ABIS online – Advanced Bacterial Identification Software, an original tool for phenotypic bacterial identification. Regnum Prokaryotae.
  60. https://www.tgw1916.net/bacteria_Bacillus_input.php
  61. Tran, L. H., & Nagano, H. (2002). Isolation and characteristics of Bacillus subtilis CN2 and its collagenase production. Journal of Food Science, 67(3), 1184-1187.
  62. https://doi.org/10.1111/j.1365-2621.2002.tb09474.x
  63. Wanapat, M., Kang, S., & Phesatcha, K. (2013). Enhancing buffalo production efficiency through rumen manipulation and nutrition. Buffalo Bulletin, 32(1), 258-275.
  64. https://kukrdb.lib.ku.ac.th/journal/BuffaloBulletin/search_detail/result/286523
  65. Watanabe, K. (2004). Collagenolytic proteases from bacteria. Applied Microbiology and Biotechnology, 63, 520-526.
  66. https://link.springer.com/article/10.1007/s00253-003-1442-0
  67. Zhao, G.Y., Zhou, M. Y., Zhao, H. L., Chen, X. L., Xie, B. B., Zhang, X. Y., He, H. L., Zhou, B. C., & Zhang, Y. Z. (2012). Tenderization effect of cold-adapted collagenolytic protease MCP-01 on beef meat at low temperature and its mechanism. Food Chemistry, 134, 1738-1744.
  68. https://doi.org/10.1016/j.foodchem.2012.03.118
  69. Ziauddin, S. K., Mahendrakar, N. S., Rao, D. N., Ramesh, B. S., & Aml, A, B. L. (1994). Observations on some chemical and physical characteristics of buffalo meat. Meat Science, 37, 103-113.
  70. https://doi.org/10.1016/0309-1740(94)90148-1