RECUPERAÇÃO DE ENZIMAS FIBRINOLÍTICAS E COLAGENOLITÍCAS DE RESÍDUOS DE PEIXES E CAMARí-ES: FONTE POTENCIAL PARA APLICAÇÃ-ES BIOMÉDICAS
DOI:
https://doi.org/10.20950/1678-2305.2019.45.1.389Palavras-chave:
subprodutos, colagenase, trombolítica, resíduoResumo
As indústrias de peixe e camarão geram uma quantidade significativa de subprodutos. Estes subprodutos podem ser utilizados para a extração de enzimas de interesse biomédico, como fibrinolítica e o colagenolítica. Assim, este trabalho teve como objetivo realizar uma triagem de subprodutos de peixes e camarões como fontes de enzimas com atividade fibrinolítica e colagenolítica e caracterizar as propriedades bioquímicas do extrato bruto com atividade colagenolítica a partir de resíduos de Cichla ocellaris. Enzimas fibrinolíticas foram recuperadas com atividades entre 5,51 ± 0,02 U.mL-1 (Caranx crysos) e 56,16 ± 0,42 U.mL-1 (Litopenaeus vannamei), enquanto enzimas colagenolitícas foram detectadas numa variação entre 6,79 ± 0,00 U.mg-1 (Trachurus lathami) e 94,35 ± 0,02 U.mg-1 (C. ocellaris). Após a triagem colagenolítica, a espécie selecionada foi a C. ocellaris, sendo submetida a um pré-aquecimento, o que culminou com um aumento de atividade enzimática de 35.07% (127.44 ± 0.09 U.mg-1). A atividade colagenolítica ótima recuperada de resíduos de C. ocellaris foi a 55°C (termoestável entre 25 a 60°) e 7,5 (estável entre 6,5 a 11,5) para avaliações de temperatura e de pH, respectivamente. Os parí¢metros cinéticos foram determinados, obtendo-se Km de 5,92 mM e Vmax de 294,40 U.mg-1. A enzima recuperada foi sensível aos íons de Cu2+, Hg2+ e Pb2+, sendo parcialmente inibida por Fluoreto de Fenilmetilsulfonil (PMSF), N-α-tosil-L-lisina clorometil cetona (TLCK) e Benzamidina. Ainda, foi capaz de clivar o colágeno nativo tipo I, o tipo mais desejável pela indústria. Assim, a recuperação de biomoléculas, além de oferecer í indústria uma fonte alternativa de moléculas ativas, contribui para a redução do impacto ambiental, agregando valor ao produto pesqueiro e proporcionando nova fonte de renda.
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Amiral, J.; Seghatchian, J. 2017. Monitoring of anticoagulant therapy in cancer patients with thrombosis and the usefulness of blood activation markers. Transfusion and Apheresis Science, 56(3): 279-286. https://doi.org/10.1016/j.transci.2017.05.010
Barbosa, R.G.; Borghesan, A.C.; Cerqueira, N.F.; Hussni, C.A.; Alves, A.L.G.; Nicoletti, J.L.M.; Fonseca, B.P.A. 2009. Fisiopatologia da trombose da veia jugular em equinos: revisão. Veterinária e Zootecnia, 16(1): 26-37.
Behl, T.; Velpandian, T.; Kotwani, A. 2017. Role of altered coagulation-fibrinolytic system in the pathophysiology of diabetic retinopathy. Vascular Pharmacology, 92(5): 1-5.
Bezerra, R.S.; Buarque, D.S.; Amaral, I.P.G.; Castro, P.F.; Espósito, T.S.; Carvalho JR, L.B. 2006. Propriedades e aplicações biotecnológicas das proteases de vísceras de peixes. In: Cyrino, J.E.P.; Urbinati, E.C. (Eds.). AquaCiência 2004 Tópicos Especiais em Biologia Aquática e Aquicultura. Artpoint Produção Gráfica, Campinas. p. 261 í 275.
Byun, H.G.; Park, P.J.; Sung, N.J.; Kim, S.K. 2002. Purification and characterization of a serine proteinase from the tuna pyloric caeca. Journal of Food Biochemistry, 26(6): 479í 494.
Cahu, T.B.; Santos, S.D.; Mendes, A.; Córdula, C.R.; Chavante, S.F.; Carvalho, L.B.; Nader, H.B.; Bezerra, R.S. 2012 Recovery of protein, chitin, carotenoids and glycosaminoglycans from pacific white shrimp (Litopenaeus vannamei) processing waste. Process Biochemistry, 47(4): 570í 577. http://dx.doi.org/10.1016/j.procbio.2011.12.012
Chandrashekar, A.; Singh, G.; Garry, J.; Sikalas, N.; Labropoulos, N. 2018. Mechanical and biochemical role of fibrin within a venous thrombus. European Journal of Vascular and Endovascular Surgery, 55(3): 417-424. http://dx.doi.org/10.1016/j.ejvs.2017.12.002
Chen, J.; Li, L.; Yi, R.; Gao, R.; Ele, J. 2018. Release kinetics of Tilapia scale collagen I peptides during tryptic hydrolysis. Food Hydrocolloids, 77(4): 931-936.
Chu, Q.; Lopez, M.; Hayashy, K.; Lonescu, M.; Billinghurst, R.C.; JohnsoN, K.A., Poole A.R.; Markel, M.D. 2002. Elevation of a collagenase generated type II collagen neoepitope and proteoglycan epitopes in synovial fluid following induction of joint instability in the dog. Osteoarthritis Cartilage, 10(8): 662-669. http://dx.doi.org/10.1053/joca.2002.0812
Cicha, I. 2015. Thrombosis: novel nanomedical concepts of diagnosis and treatment. World Journal of Cardiology, 7(8): 434-441. http://dx.doi.org/10.4330/wjc.v7.i8.434.
Daboor, S.M.; Budge, S.M.; Ghaly, A.E.; Brooks, M.S.; Dave, D. 2010. Extraction and Purification of Collagenase Enzymes: A Critical Review. American Journal of Biochemistry and Biotechnology, 6(4): 239-263.
Daboor, S.M.; Budge, S.M.; Ghaly, A.E.; Brooks, M.S.; Dave, D. 2012. Isolation and activation of collagenase from fish processing waste. Advances in Bioscience and Biotechnology, 3(3): 191í 203.
Fayyad, A.; Lapp, S.; Risha, E.; Pfankuche, V.M.; Rohn, K.; Barthel, Y.; Schaudien, D.; Baumgí¤rtner, W.; Puf, C. 2018. Matrix metalloproteinases expression in spontaneous canine histiocytic sarcomas and its xenograft model. Veterinary Immunology and Immunopathology, 198(4): 54í 64. http://dx.doi.org/10.1016/j.vetimm.2018.03.002
Freitas-Júnior, A.C.V.; Costa, H.M.S.; Icimoto, M.Y.; Hirata, I.Y.; Marcondes, M.; CarvalhO, L.B.; Oliveira. V.; Bezerra, R.S. 2012. Giant Amazonian fish pirarucu (Arapaima gigas): Its viscera as a source of thermostable trypsin. Food Chemistry, 133(4): 1596-1602. http://dx.doi.org/10.1016/j.foodchem.2012.02.056
Guedes, A.F.; Carvalho, F.A.; Domingues, M.M.; Macrae, F.L.; McPherson, H.R.; SantoS, N.C.; AriÑ"˜ns, R.A.S. 2018. Sensing adhesion forces between erythrocytes and γ’ fibrinogen, modulating fibrin clot architecture and function. Nanomedicine: Nanotechnology, Biology and Medicine, 14(3): 909-918. http://dx.doi.org/10.1016/j.nano.2018.01.006
Hayashi, K.; Kim, S.Y.; Lansdowne, J.L.; Kapatkin, A.; Déjardin, L.M. 2009. Evaluation of a collagenase generated osteoarthritis biomarker in naturally occurring canine cruciate disease. Veterinary Surgery, 38(1): 117-21.
Hayet, B.K.; Rym, N.; Ali, B.; Sofiane, G.; Moncef, N. 2011. Low molecular weight serine protease from the viscera of sardinelle (Sardinella aurita) with collagenolytic activity: Puriï¬Âcation and characterization. Food Chemistry, 124(3): 788í 794.
Hernandez-herrero, M.M.; DufloS, G.; Malle, P.; Bouquelet, S. 2003. Collagenase activity and protein hydrolysis as related to spoilage of iced cod (Gadus morhua). Food Research International, 36(2): 141í 147. http://dx.doi.org/10.1016/S0963-9969(02)00129-1
Jayes, F.L.; Liu, B.; Moutos, F.T.; Kuchibhatla, M.; Guilak, F.; leppert, P.C. 2016. Loss of stiffness in collagen-rich uterine fibroids after digestion with purified collagenase Clostridium histolyticum. American Journal of Obstetrics and Gynecology, 215(5): 596.e1í 596.e8. http://dx.doi.org/10.1016/j.ajog.2016.05.006ur
Karakurt, A.; BaÅŸbuÄŸ, H.S. 2015. Prosthetic heart valve thrombosis treated with low-dose slow-infusion fibrinolytic therapy. Journal of Cardiology Cases, 12(1): 12-15.
Kim, S. K.; Park, P. J.; Kim, J.B.; Shahidi, F. 2002. Purification and characterization of a collagenolytic protease from the filefish, Novoden modestrus. Journal of biochemistry and molecular biology, 35(2): 165-171.
Kristjánsson, M. M.; Gudmundsdóttir S.; Fox, J. W.; Bjarnason, J. B. 1995. Characterization of collagenolytic serine proteinase from the Atlantic cod (Gadus morhua). Comparative Biochemistry and Physiology - Part B, 110(4): 707í 717.
Leite, S.B.P.; Sucasas, L.F.A.; Oetterer, M. 2016. Resíduos da comercialização de pescado marinho í volume de descarte e aspectos microbiológicos. Revista Brasileira de Tecnologia Agroindustrial, 10(1): 2112-2125.
Lima, C.A.; Campos, J.F.; Lima Filho, J.L.; Converti, A.; Carneiro da Cunha, M.G.; Porto, A.L.F. 2014. Antimicrobial and radical scavenging properties of bovine collagen hydrolysates produced by Penicillium aurantiogriseum URM 4622 collagenase. Journal of Food Science and Technology, 52(7): 4459í 4466. http://dx.doi.org/10.1007/s13197-014-1463-y
Lima-Junior, E.M.; Picollo, N.S.; Miranda, M.J.B.; Ribeiro, W.L.C.; Alves, A.P.N.N.; Ferreira, G.E.; Parente, E.A.; Moraes Filho, M.O. 2017. Uso da pele de tilápia (Oreochromis niloticus), como curativo biológico oclusivo, no tratamento de queimaduras. Revista Brasileira de Queimaduras, 16(1): 10-17.
Moore, S.; Stein, W. 1954. A modified ninhydrin reagent for the photometric determination of amino acids and related compounds. The Journal of Biological Chemistry, 211(2): 907í 913.
Mukherjee, J.; Webster, N.; Llewellyn, L.E. 2009. Purification and Characterization of a Collagenolytic Enzyme from a Pathogen of the Great Barrier Reef Sponge, Rhopaloeides odorabile. PLoS ONE, 4(9): 1í 5.
Murado, M.A.; González, M.P.; Vázquez, J.A. 2009. Recovery of proteolytic and collagenolytic activities from viscera by-products of Rayfish (Raja clavata). Mar Drugs, 7(4): 803í 815. http://dx.doi.org/10.3390/md7040803
Oliveira, V.M.; Assis, C.R.D.; Herculano, P.N.; Cavalcanti, M.T.H.; Bezerra, R.S.; Porto, A.L.F. 2017a. Collagenase from smooth weakfish: extraction, partial purification, characterization and collagen specificity test for industrial application. Boletim do Instituto de Pesca, 43(1): 52í 64. http://dx.doi.org/10.20950/1678-2305.2017v43n1p52
Oliveira, V.M., Carneiro Cunha, M.N.; Assis, C.R.D.; Nascimento, T.P.; HerculanO, P.N.; Cavalcanti, M.T.H.; Porto, A.L. 2017b. Colagenases de pescado e suas aplicações industriais. Pubvet, 11(3): 243-255. http://dx.doi.org/10.22256/pubvet.v11n3.243-255
Oliveira, V.M., Carneiro Cunha, M.N.; Nascimento, T.P.; Assis, C.R.D.; Bezerra, R.S.; Porto, A.L.F. 2017c. Collagen: general characteristics and production of bioactive peptides - a review with emphasis on byproducts of fish. ActaFish, 5(2): 70-82. http://dx.doi.org/10.2312/ActaFish.2017.5.2.70-82
Oliveira, V.M.; Nascimento, T.P.; Assis, C.R.D.; Bezerra, R.S.; Porto, A.L.F. 2017d. Study on enzymes of industrial interest in digestive viscera: Greater amberjack (Seriola dumerili). Journal of Coastal Life Medicine, 5(6): 233-238. http://dx.doi.org/10.12980/jclm.5.2017J6-300
Park, J.P.; Lee, S.H.; Byun, H.G.; Kim, S.H.; Kim, S.K. 2002. Purification and characterization of a collagenase from the mackerel, Scomber japonicus. Journal of biochemistry and molecular biology, 35(6): 576í 582.
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2019-02-13
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