Produção e caracterização de protease fibrinolítica de Streptomyces parvulus DPUA 1573

Abstract

 RESUMOAs proteases fibrinolíticas são capazes de degradar coágulos de fibrina formados dentro dos vasos sanguíneos, evitando a trombose intravascular. Em animais, a tromboflebite, que acomete frequentemente os equinos, ocasiona, em seus casos graves, a obstrução jugular e também um edema de laringe, derivando a obstrução das vias aéreas, o que possibilita um edema cerebral, ocorrendo o óbito do animal. Devido ao fato de o tratamento ser de custo elevado, faz-se necessária a investigação de outras fontesde proteases fibrinolíticas com custos menores e com menos efeitos colaterais. Diante disso, este estudo tem como objetivo produzir e caracterizar proteases fibrinolíticas obtidas de Streptomyces parvulus DPUA 1573. Para produção da enzima, foi utilizado um planejamento fatorial 2 4 avaliando a concentração da farinha de soja (0,5, 1,0 e 1,5%) e da glicose (0, 0,5 e 1,0g/L), temperatura (28, 32 e 37ºC) e agitação (150, 200 e 250rpm) sobre a biomassa e a atividade fibrinolítica. Pode-se verificar que a protease fibrinolítica apresentou atividade máxima (835U/mL) nas condições de concentração de 1,5% de soja, 1g/L de glicose, 28°C e 150rpm com 48 horas de fermentação. A protease fibrinolítica obtida teve temperatura e pH ótimos de 55°C e pH 9,0, respectivamente. A atividade enzimática foi inibida pelo EDTA, pelo íon Fe 2+ e pelo SDS, o que indicou a enzima ser uma metaloprotease. A linhagem Streptomyces parvulus DPUA 1573 foi capaz de produzir protease fibrinolítica, possuindo características bioquímicas favoráveis à aplicação na medicina veterinária e possivelmente humana. Palavras-chave: Streptomyces parvulus, proteases fibrinolíticas, fermentação submersa ABSTRACT Fibrinolytic proteases are able to degrade fibrin clot formed in the blood vessel, avoiding intravascular thrombosis. In animals, thrombophlebitis often affects horses, and in severe cases causes obstruction of the jugular and laryngeal edema leading to airway obstruction allowing cerebral edema resulting in the death of the animal. Since treatment is costly, the investigation of other sources of fibrinolytic proteases at lower costand with fewer side effects is needed. Thus, this study aims to produce and characterize fibrinolytic proteases from Streptomyces parvulus DPUA 1573. For enzyme production, a factorial design was performed to evaluate 2 4 soybean flour concentration (0.5, 1.0 and 1.5%) and glucose (0, 0.5 and 1.0g/L), temperature (28, 32 and 37°C) and agitation (150, 200 and 250rpm)

Serine metalloprotease with plasmin-like fibrinolytic activity of Serratia marcescens isolated from the Amazon basin

“…The enzyme is highly stable at room temperature over a wide pH range, maintaining more than 80% of its activity in the pH range 5.0 to 9.0 for a 24 hour period. The alkaline pH was also reported to provide better results for the Brazilian Journal of Development, Curitiba, v.9, n.6, p. 20249-20268, jun., 2023 fibrinolytic enzymes of Streptomyces parvulus [29], S. marcescens RSPB11 [14] and Bacillus subtilis ICTF-1 [30] which showed optimal activity at pH 9. Another fibrinolytic enzyme produced by Bacillus subtilis, showed greater activity at pH 7.0, and was relatively stable between pH 6.0-10.0 [31].…”

Purification and characterization of fibrinolytic protease from Streptomyces parvulus by polyethylene glycol-phosphate aqueous two-phase system

“…The Amazon holds an inestimable genetic resources bank, allowing the attainment of a varied products range, mainly biotechnological products from microorganisms (Bittencourt & Rios 2013). Streptomyces parvulus DPUA 1573 isolated from Amazonian lichens was studied by Batista et al 2017, showing promise to produce fibrinolytic enzymes. However, studies are still needed on alternatives for the purification of the enzyme from the crude extract and its characterization.…”