STUDY OF OPTIMIZATION, CHARACTERIZATION AND APPLICATIONS OF KERATINASE PRODUCED BY A BACILLUS STRAIN

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Authors

  • Arjuman Surti
  • Mugdha Taral

Keywords:

Bacillus sp. AM251, feather, keratinase, keratin, poultry, optimization

Abstract

The poultry industry contributes to immense amounts of feather waste, over 90% of which is keratin. Although these industrial byproducts can be used as animal feed due to their high nutritional value, they are often landfilled or composted due to their low decomposition. The present study was carried out to isolate and identify a potential keratinase-producing bacterium from soil and waste water samples and to optimize cultural and physicochemical conditions to maximize enzyme production. The bacterium with the highest enzyme activity among the 17 isolates obtained in our study was identified as Bacillus sp. AM251. Optimal enzyme production was observed in media with the following composition (in g/L); NaCl (0.5), KH2PO4 (0.7), K2HPO4 (1.4), MgSO4 (0.01), CaCl2 (0.5), ammonium sulphate (2.5) and feather (10); pH 7.0. The optimum physicochemical parameters were determined to be 7.5% inoculum size, 37°C incubation temperature and shaking conditions (200 rpm). Characterization studies showed that the enzyme is a 43kDa serine protease with optimum activity at 50°C and pH 8.0. As an application of the enzyme keratinase, we also studied its substrate specificity and depilatory effect on goat skin. Both studies showed good enzyme activity. However, another study conducted to determine the use of enzyme-treated feather meal as an alternative to yeast extract and peptone for the growth of microorganisms in the laboratory showed limited applications. Overall, the results of our study show that the strain Bacillus sp. AM251 has great potential as a keratinase producer that can be used for bioremediation of feathers in landfills, preparation of animal feed or as a depilatory agent.

References

Bragulla, H. H., Homberger, D. G. (2009): Structure and functions of keratin proteins in simple, stratified, keratinized and cornified epithelia. Journal of Anatomy 214(4): 516–559.

Moll, R., Divo, M., Langbein, L. (2008): The human keratins: biology and pathology. Histochemistry and Cell Biology 129(6): 705–733.

Qiu, J., Wilkens, C., Barrett, K., Meyer, A. S. (2020): Microbial enzymes catalyzing keratin degradation: Classification, structure, function. Biotechnology Advances 44: 107607.

Paul, T., Mandal, A., Mondal, K. C. (2018): Waste to value aided fertilizer: an alternative cleaning technique for poultry feathers waste disposal. Annals of Microbiology and Immunology 1(2): 1006.

Lange, L., Huang, Y., Busk, P. K. (2016): Microbial decomposition of keratin in nature- a new hypothesis of industrial relevance. Applied Microbiology and Biotechnology 100(5): 2083–2096.

Franke- Whittle, I. H., Insam, H. (2013): Treatment alternatives of slaughterhouse wastes, and their effect on the inactivation of different pathogens: a review. Critical Reviews in Microbiology 39(2): 139–151.

Woo, Y. K., Kim, J. H. (2006): Fowl cholera outbreak in domestic poultry and epidemiological properties of Pasteurella multocida isolate. Journal of Microbiology (Seoul Korea) 44(3): 344–353.

Yamamoto, Y., Nakamura, K., Yamada, M., Mase, M. (2010): Persistence of avian influenza virus (H5N1) in feathers detached from bodies of infected domestic ducks. Applied and Environmental Microbiology 76(16): 5496–5499.

Kumar, J. (2021): Microbial hydrolysed feather protein as a source of amino acids and protein in the diets of animals including poultry. In (Ed.) Advances in Poultry Nutrition Research. IntechOpen. https://doi.org/10.5772/intechopen.96925

Vidmar, B., Vodovnik, M. (2018): Microbial Keratinases: Enzymes with promising biotechnological applications. Food Technology and Biotechnology 56(3): 312–328.

Peng, Z., Mao, X., Zhang, J., Du, G., Chen, J. (2020): Biotransformation of keratin waste to amino acids and active peptides based on cell-free catalysis. Biotechnology for Biofuels 13: 61.

Lagos, J. E., Intodia, V. (2016): Market development reports on livestock and products. Hides and Skins - 2016 Global Agricultural Information network. GAIN Report Number IN6078. Available at: https://agriexchange.apeda.gov.in/MarketReport/Reports/Hides_Skins%20-%202016_New%20Delhi_India_6-9-2016.pdf (Accessed 20th October 2020).

Mokrejš, P., Huťťa, M., Pavlačková, J., Egner, P. (2017): Preparation of keratin hydrolysate from chicken feathers and its application in cosmetics. Journal of Visualized Experiments: JoVE (129): 56254.

Hassan, M. A., Haroun, B. M., Amara, A. A., Serour, E. A. (2013): Production and characterization of keratinolytic protease from new wool degrading Bacillus species isolated from Egyptian ecosystem. BioMed Research International 2013: 175012.

Cai, C. G., Chen, J. S., Qi, J. J., Yin, Y., Zheng, X. D. (2008): Purification and characterization of keratinase from a new Bacillus subtilis strain. Journal of Zhejiang University. Science B 9(9): 713–720.

Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951): Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry 193(1): 265–275.

Bergey's Manual of Determinative Bacteriology (7th ed.). (1964): American Journal of Public Health and the Nations Health 54(3): 544.

Barman, N. C., Zohora, F. T., Das, K. C., Mowla, M. G., Banu, N. A., Salimullah, M., Hashem, A. (2017). Production, partial optimization and characterization of keratinase enzyme by Arthrobacter sp. NFH5 isolated from soil samples. AMB Express 7(1): 181.

Ramnani, P., Gupta, R. (2004): Optimization of medium composition for keratinase production on feather by Bacillus licheniformis RG1 using statistical methods involving response surface methodology. Biotechnology and Applied Biochemistry 40(2): 191–196.

Mazotto, A. M., Coelho, R. R., Cedrola, S. M., de Lima, M. F., Couri, S., Paraguai de Souza, E., Vermelho, A. B. (2011): Keratinase production by three Bacillus spp. using feather meal and whole feather as substrate in a submerged fermentation. Enzyme Research 2011: 523780.

Suharti, D. R., Tyas., and Nilamsari, N. R. (2019): Isolation and characterization of a newly keratinase producing Bacillus sp. N1 from tofu liquid waste. IOP Conference Series: Earth and Environmental Science, International Conference on Green Agro-industry and Bioeconomy 18–20 September 2018, Universitas Brawijaya, East Java Indonesia, 230:012088.

Nnolim, N. E., Udenigwe, C. C., Okoh, A. I., Nwodo, U. U. (2020): Microbial keratinase: next generation green catalyst and prospective applications. Frontiers in Microbiology 11: 580164.

Zhang, R. X.., Wu, Z. W., Cui, H. Y., Chai, Y. N., Hua, C. W., Wang, P., Li, L., Yang, T. Y. (2022): Production of surfactant-stable keratinase from Bacillus cereus YQ15 and its application as detergent additive. BMC Biotechnology 22: 26.

Prasad, H. V., Kumar, G., Karthik, L., Rao, K. V. B. (2010): Screening of extracellular keratinase producing bacteria from feather processing areas in Vellore, Tamil Nadu, India. Journal of Scientific Research 2(3): 559-565.

Subugade, S., Gupta, S. G., Mokashe, S. (2017): Isolation and screening of keratinase producing bacteria from chicken feather dumping site. International Journal of ChemTech Research 10(5): 900-905.

Raju, E. V. N., Divakar, G. (2013): Screening and isolation of keratinase producing bacteria from poultry waste. International Journal of Pharmaceutical Research & Allied Sciences 2(1): 70-74.

Shen, N., Yang, M., Xie, C., Pan, J., Pang, K., Zhang, H., Wang, Y., Jiang, M. (2022): Isolation and identification of a feather degrading Bacillus tropicus strain Gxun-17 from marine environment and its enzyme characteristics. BMC Biotechnology 22: 11.

Karadaglibahri, M., Ozcan, D. (2022): Isolation of keratinase-producing Bacillus strains and enhanced enzyme production using in vitro mutagenesis. Cellular and molecular biology 94 (1): 2022.

Sekar, V., Kannan, M., Ganesan, R. Dheeba, B., Sivakumar, N., Kannan, K. (2016): Isolation and screening of keratinolytic bacteria from feather dumping soil in and around Cuddalore and Villupuram, Tamil Nadu. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences 86: 567–575.

Martindro, R. L., Wibowo, S. A., Bachruddin, Z., Fitriyanto, N. A., Nakagawa, T., Hayakawa, T., Pertiwiningrum, A. (2017): Screening and characterization of keratinolytic bacteria from Puffer fish skin waste. Pakistan Journal of Nutrition 16: 488-496.

Buchanan, R. E., Gibbons, N. E. (1974): Bergey's Manual of Determinative Bacteriology. 8th Edition, Williams and Wilkins, Baltimore, pp. 1268.

Ahmed, M., Arshad, Y., Mukhtar, H. (2019): Isolation and screening of keratinase producing bacteria from soil. Biologia – Journal of Biological Society of Pakistan 65(2): 1-6.

Ningthoujam, D. S., Jaya, L., Devi, P. J., Kshetri, P., Tamreihao, K., Mukherjee, S., Devi, S. S., Nongthombam, B. (2016): Optimization of keratinase production by Amycolatopsis sp. Strain MBRL 40 from a Limestone Habitat. Journal of Bioprocess and Biotechnology 6: 282.

Nurliyana, M., Lai-yee, P., Suraini, A. A. (2017): Optimization of metallo-keratinase production by Pseudomonas sp. LM19 as a potential enzyme for feather waste conversion. Biocatalysis and Biotransformation 35(1): 41-50.

Moonnee, Y. A., Foysal, M. J., Hashem, A., Miah, M. F. (2021): Keratinolytic protease from Pseudomonas aeruginosa for leather skin processing. Genetic Engineering & Biotechnology 19(1): 53.

Razzaq, A., Shamsi, S., Ali, A., Ali, Q., Sajjad, M., Malik, A., Ashraf, M. (2019): Microbial Proteases Applications. Frontiers in Bioengineering and Biotechnology 7: 110.

Sharma, M., Gat, Y., Arya, S., Kumar, V., Panghal, A., Kumar, A. (2019): A review on microbial alkaline protease: an essential tool for various industrial approaches. Industrial Biotechnology 15(2): 69-78.

Mazotto, A. M., Cedrola, S. M. L., de Souza, E. P., Couri, S., Vermelho, A. B. (2022): Enhanced keratinase production by Bacillus subtilis amr using experimental optimization tools to obtain feather protein lysate for industrial applications. 3 Biotech 12(4): 90.

Lin, X., Wong, S. L., Miller, E. S., Shih, J. C. (1997): Expression of the Bacillus licheniformis PWD-1 keratinase gene in B. subtilis. Journal of Industrial Microbiology and Biotechnology 19(2): 134-138.

Gurav, R. G., Jadhav, J. P. (2013): Biodegradation of keratinous waste by Chryseobacterium sp. RBT isolated from soil contaminated with poultry waste. Journal of Basic Microbiology 53: 128–135.

Fakhfakh, N., Kanoun, S., Manni, L., Nasri, M. (2009): Production and biochemical and molecular characterization of a keratinolytic serine protease from chicken feather-degrading Bacillus licheniformis RPk. Canadian Journal of Microbiology 55: 427–436.

de Paiva, D. P., de Oliveira, S. S., Mazotto, A. M., Vermelho, A. B., de Oliveira, S. S. (2019): Keratinolytic activity of Bacillus subtilis LFB-FIOCRUZ 1266 enhanced by whole-cell mutagenesis. 3 Biotech 9(1): 2.

Jeevana, L. P., Kumari, C. C., Lakshmi, V. V. (2013): Efficient degradation of feather by keratinase producing Bacillus sp. International Journal of Microbiology 2013: 608321.

Nagal, S., & Jain, P. C. (2010): Feather degradation by strains of Bacillus isolated from decomposing feathers. Brazilian Journal of Microbiology 41(1): 196–200.

Corrêa, A. P. F., Daroit, D. J., Brandelli, A. (2010): Characterization of a keratinase produced by Bacillus sp. P7 isolated from an Amazonian environment. International Biodeterioration & Biodegradation 64(1): 1-6.

Thys, R. C. S., Lucas, F. S., Riffel, A., Heeb, P., Brandelli, A. (2004): Characterization of a protease of a feather-degrading Microbacterium species. Letters Applied Microbiology 39(2): 181–186.

News article by Globe Newswire. (2021): Global Industrial Enzymes Market Size & Share Witnessing Consistent Growth—Projected to Reach worth USD 9.2 Billion by 2027 BlueWeave Consulting and Research Pvt Ltd. Available at https://www.globenewswire.com/en/news-release/2021/10/12/2312804/0/en/Global-Industrial-Enzymes-Market-Size-Share-Witnessing-Consistent-Growth-Projected-to-Reach-worth-USD-9-2-Billion-by-2027-BlueWeave.html (Accessed 9th November 2022).

Macedo, A. J., da Silva, W. O. B., Gava, R., Driemerier, D., Henriques, J. A. P., Termignoni, C. (2005): Novel keratinase from Bacillus subtilis S14 exhibiting remarkable dehairing capabilites. Applied Environmental Microbiology 71(1): 594–596.

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Published

2024-06-21

How to Cite

Surti, A., & Taral, M. (2024). STUDY OF OPTIMIZATION, CHARACTERIZATION AND APPLICATIONS OF KERATINASE PRODUCED BY A BACILLUS STRAIN. Journal of Applied Biological Sciences, 18(2), 143–161. Retrieved from https://jabsonline.org/index.php/jabs/article/view/1161

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