First report of collagenase production by a fungus isolated from the Caatinga soil - Rhizopus microsporus UCP 1296
DOI:
https://doi.org/10.33448/rsd-v11i12.34618Keywords:
Collagenase; Filamentous fungus; Bioprocesses; Innovation.Abstract
The present work aimed to study the fungus Rhizopus microsporus UCP 1296 isolated from the Caatinga soil, emphasizing the development and application of biotechnological processes for the production of a new collagenase. Therefore, significant fermentation parameters such as temperature, pH, culture medium and fermentation time were evaluated. Samples of the microorganism were kept at 4 °C in malt extract agar medium. The mycelium was suspended in 5 mL of a solution composed of NaCl and Tween 80. After inoculation with the spore suspension containing 150 µL (106 spores/mL), submerged fermentation was carried out in Erlenmeyer flasks containing 250 mL of gelatin. A growth curve was developed to identify the best time for enzyme production. The results show that collagenase production occurred with 120 hours of fermentation, obtaining 550 U/mL of collagenolytic activity and 0.42 g/L biomass in the stationary phase, indicating that the collagenase produced is a secondary metabolite. A 24 factorial design was carried out to select the best parameters where the influence of the variables was evaluated: temperature (ºC), pH, agitation and substrate concentration (gelatin). As a result of the factorial design, we obtained an increase in enzyme production of about 32%. This is the first study to describe optimized conditions for collagenase production by R. microsporus strains.
References
Abidi, F., Aissaoui, N., Gaudin, J. C., Chobert, J. M., Haertlé, T., & Marzouki, M. N. M. S. Analysis and molecular characterization of Botrytis cinerea protease Prot-2. Use in bioactive peptides production. Appl Biochem Biotechnol. 2013 May; 170(2):231-47.
Agrawal, D., Patidar, P. Banerjee, T., & Patil, S (2004). Production of alkaline protease by Penicillium sp. under SSF conditions and its application to soy protein hydrolysis. Process Biochem. 39(8): 977-981.
Azevedo, T. O. M., Silva, B., Lima, E. S., & Souza, J. V. B. de (2018). Fungos isolados de amostras de solo da região amazônica, Aspergillus INPA 154 e Fusarium INPA 6, produzem proteases com potencial para utilização em formulações cosméticas. Dissertação de mestrado.
Baehaki, A., Sukamo, S. D., Setyahadi, S., & Suhartono, M.T. (2014). Production and characterization of collagenolytic protease from Bacillus licheniformis F11.4 originated from Indonesia. Asian J. Chem, 26 (10): 2861-2864. 10.14233/ajchem.2014.15863
Barros Neto, B., Scarminio, I. S., & Bruns, R. E. (1996). Planejamento e Otimização de Experimentos. Campinas: Ed. UNICAMP, 2: 299 ISBN: 8526803360
Benabda, O., M’hir, S., Kasmi, M., Mnif, W., & Hamdi, M. (2019). Optimization of Protease and Amylase Production by Rhizopus oryzae Cultivated on Bread Waste Using Solid-State Fermentation. Hindawi Journal of Chemistry. 3:1-9. 10.1155/2019/3738181
Bhagwat, P. K., & Dandge, P. B. (2018). Colágeno e proteases colagenolíticas: Uma revisão. Biocatálise e Biotecnologia Agrícola. 15 (18), 43-55.
Chavira, R. J., Burnett, T. J., & Hageman, J. H. (1984). Assaying proteinases with azocoll. Anal. Biochem, 136:4446–4450.10.1016/0003-2697(84)90242-2.
Cui, Y. Q., Van der Lans, R. G. J. M., & Luyben, K. (1998). Effects of dissolved oxygen tension and mechanical forces on fungal morphology in submerged fermentation. Biotechnology and Bioengineering. 57(4):409-419.
Daboor, S. M., Budge, S. M., Ghaly, A. E., Brooks, S., & Dave, D. (2010). Extraction and Purification of Collagenase Enzymes: A Critical Review. American Journal of Biochemistry and Biotechnology, 6(4): 239-263. 10.3844/ajbbsp.2010.239.263
Da Rocha, W. R. V., Agamez-Montalvo, G. S., Feitosa,V. A., Machado, S. E. F., De Souza Lima, G. M., Pessoa-Jr, A., & Alves, H. S. (2019). Screening and optimizing fermentation production of lasparaginase by Aspergillus terreus strain S-18 isolated from the Brazilian Caatinga Biome. Journal of Applied Microbiology. 126(5): 1426–1437. 10.1111/jam.14221
EL-Enshasy, H., Kleine, J., & Rinas, U. (2006). Agitation effects on morphology and protein productive fractions of filamentous and pelleted growth forms of recombinant Aspergillus niger. Process Biochemistry. 41: 2103-2112: 10.1016/j.procbio.2006.05.024
Elibol, M., & Moreira, A.R. (2005). Optimizing some factors affecting alkaline protease production by a marine bacterium Teredinobacter turnirae under solid substrate fermentation. Process biochemistry. 40 (5), 1951-1956:10.1016/j.procbio.2004.07.010
Gao, S., Hong, H., Zhang, C., Wang, K., & Zhang, B. (2019). Immunomodulatory effects of collagen hydrolysates from yak (Bos grunniens) bone on cyclophosphamide-induced immunosuppression in BALB/c mice. Journal of Functional Foods. 60 (1)1034-40: 10.1016/j.jff.2019.103420
Gupta, R., Gupta, N., & Rathi, P. (2004). Bacterial lipases: an overview of production, purification and biochemical properties. Appl Microbiol Biotechnol. 64(6):763-81: 10.1007/s00253-004-1568-8.
Gurumallesh, P., Alagu, K., Ramakrishnan, B., & Muthusamy, S. (2019). A systematic reconsideration on proteases. International Journal of Biological Macromolecules. 128: 254–267: 10.1016/j.ijbiomac.2019.01.081.
Graminho, E. R., Da Silva, R. R., De Freitas Cabral, T. P., Arantes, E. C., Da Rosa, N. G., Juliano, L., Okamoto, D. N., De Oliveira, L. C., Kondo, M. Y., Juliano, M. A. & Cabral, H. (2013). Purification, characterization, and specificity determination of a new serine protease secreted by Penicillium waksmanii. Appl Biochem Biotechnol, 169(1): 201– 214: 10.1007/s12010-012-9974-3.
Hasan, F., Shah, A. A., & Hammed, A. (2006). Industrial application of microbial lipases. Enzyme and Microbial Technology, 39 (2): 235-251: 10.1016/j.enzmictec.2005.10.016
IBGE - Instituto Brasileiro de Geografia e Estatística: Dados gerais do município, 2015. < http://cod.ibge.gov.br/N1G4 >
Jain, R., & Jain, P. C. (2010). Production and partial characterization of collagenase of Streptomyces exfoliatus CFS 1068 using poultry feather. Indian J. Exp. Biol., 48(2), 174–8 http://www.ncbi.nlm.nih.gov/pubmed/20455327
Kermasha, S., & Eskin, M. N. Enzymes. In: Kermasha, S. & Eskin, M. N. (2021). Enzymes Novel Biotechnological Approaches for the Food Industry. (1a ed.): Academic Press. 2: 15-44.
Kumar & Singh, D. (2014). Global scenario of industrial enzyme market Complimentary Contributor Copy. Industrial Enzymes, 10: 173–196.
Lima, C. A., Rodrigues, P. M. B., Porto, T. S., Viana, D. A., Lima Filho, J. L., Porto, A. L. F., & Carneiro-da-Cunha, M. G. (2009). Production of a collagenase from Candida albicans URM 3622. Biochem Eng J. 43(3): 315–320:10.1016/j.bej.2008.10.014
Lima, C. A., Lima Filho, J. L., Neto, B. B., Converti, A., Carneiro-da-Cunha, M. G., & Porto, A. L. F. (2011). Production and characterization of a collagenolytic serine proteinase by Penicillium aurantiogriseum URM 4622: A factorial study. Biotechnol. Bioprocess Eng. 16 (3), 549–560:10.1007/s12257-010-0247-0
Lima, C. A., Freitas-Junior, A. C. V., Lima-Filho, J. L., Converti, A., Marques, D. A. V., Carneiro-da-Cunha, M. G., & Porto, A. L. F. (2013). Two-phase partitioning and partial characterization of a colagenase from Penicillium aurantiogriseum URM 4622: Application to collagen hydrolysis. Biochemical Engineering Journal, 75: 64-71:10.1016/j.bej.2013.03.012
Mahmoud, Y. A. G., Aabu El-Souod, S. M., El-Shourbagy, S. M., & El-Badry, A. S. M. (2007). Characterisation and inhibition effect of cetrimide on collagenase produced by Aspergillus flavus, isolated from mycotic ulcers. Ann. Microbiol., 57(1), 109– 113 doi.org/10.1007/BF03175058
Merheb, C. W., Cabral, H., & Da Silva, R. (2007). Partial characterization of protease from a thermophilic fungus, Thermoascus aurantiacus, and its hydrolytic activity on bovine casein. Food Chemistry. 104:127–131: 10.1016/j.foodchem.2006.11.010
Montgomery, D. C. (1991). Design and Analysis of Experiments. (3rd ed.,) John Wiley & Sons, Inc., Nova York.
Nigam, P. S. (2013). Microbial enzymes with special characteristics for biotechnological applications. Biomolecules. 23(3): 597–611: 10.3390/biom3030597.
Oliveira, V. M., Assis, C. R. D., Herculano, P. N., Cavalcanti, M. T. H., Bezerra, R. S., & Porto, A. L. F. (2017). Collagenase from smooth weakfish: extraction, partial purification, characterization and collagen. Boletim do Instituto de Pesca, 43(1): 52-64: 10.20950/1678-2305.2017v43n1p52
Ok, T., & Hashinaga, F. (1996). Detection and production of extracellular collagenolytic enzyme from Zygosaccharomyces rouxii. J. Gen. Appl. Microbiol., 42, 517–523: 10.2323/jgam.42.517
Pamboukian, C. R. D. (2003). Produção do antitumoral rentamicina por Streptomyces olindenses em processos descontínuos. (Doutorado em Engenharia Química). Programa de Pós Graduação em Engenharia, USP.
Sandhya, C., Sumantha, A., Szakacs, G., & Pandey, A. (2005). Comparative evaluation of neural protease production by Aspergillus oryzae in submerged and solid state fermentation. Process Biochem., 40: 2689–2694. doi.org/10.1016/j.procbio.2004.12.001
Silva, O. S., Gomes, M. H. G., Oliveira, R. L., Porto, A. L. F., Converti, A., & Porto, T. S. (2017). Partitioning and extraction protease from Aspergillus tamarii URM4634 using PEG-citrate aqueous two-phase systems. Biocatalysis and Agricultural Biotechnology. 9: 168-173.
Silva, I. E. B. da., Jardim, L. A. de S., Ribeiro, E. M. S., & Lima, R. L. F. de A. (2020). Fungolândia: Jogo educativo de tabuleiro sobre a diversidade e importância dos fungos da caatinga. Revista Brasileira de Educação Ambiental (REVBEA), 15(6): 52–99 : 10.34024/revbea.2020.v15.11449
Smith, P. K., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., Fujimoto, E. K., Goeke, N. M., Olson, B. J., & Klenk, D. C. (1985). Measurement of protein using bicinchoninic acid. Analytical Biochemistry,150(1): 76-85:10.1016/0003-2697(85)90442-7.
Sumantha, A., Deepa, P., Sandhya, C., Szakacs, G., Soccol, C. R., & Pandey, A. (2006). Rice bran as a substrate for proteolytic enzyme production. Brazilian Arch Biol Technol. 49(5):843–851. http://doi.org/10.1590/S1516-89132006000600019
Ventura, M. M., Freitas, S. M., & Freire, A. P. (2008). Catálise enzimática – alguns destaques na evolução da enzimologia In: E. P. S. Bon; M. A. Ferrara; M. L. Corvo. Enzimas em biotecnologia – Produção, Aplicações e Mercado. Rio de Janeiro: Interciência, 1: 1- 27.
Wanderley, M. C. de A., Neto, J. M. W. D., Lima Filho, J. L. de., Lima, C. A., Teixeira, J. A. C., & Porto, A. L. F. (2017). Collagenolytic enzymes produced by fungi: a systematic review. Brazilian Journal of Microbiology. 48(1): 13–24 doi.org/10.1016/j.bjm.2016.08.001.
Wanderley, M. C. de A., Neto, J. M. W. D., Lima, C. D. A., Silvério, S. I. C., Teixeira, J. Á. C., & Porto, A. L. F. (2016). Production and Characterization of Collagenase by Penicillium sp. UCP 1286 Isolated From Caatinga Soil. Journal of Applied Biology and Biotechnology, 4(4): 1-10: 10.7324/JABB.2016.40401
Wu, Q., Li, C., Chen, H., & Shuliang, L. (2010). Purification and characterization of a novel collagenase from Bacillus pumilus. J. Appl. Biochem. Biotechnol., 160 (1), 129–39:10.1007/s12010-009-8673-1
Yazid, N. A. (2017) Proteases from protein-rich waste : Production by SSF, downstream, immobilisation onto nanoparticles and application on protein hydrolysis. Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental. Tese de Doutorado. 286 p.
Zimmer, K. R., Borré, G. L., Trentin, D. S., Júnior, C. W., Frasson, A. P., Graeff, A. A., Gomes, P., & Macedo, A. J. (2009). Enzimas microbianas de uso terapêutico e diagnóstico clínico, Revista Liberato, 10(14): 123-137.
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