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

被引：6

作者：

Alencar, Viviane N. S. ^{[1
]}

Do Nascimento, Maria Clara ^{[2
]}

Dos Santos Ferreira, Julyanne, V ^{[3
]}

Da Silva Batista, Juanize M. ^{[2
]}

Da Cunha, Marcia N. C. ^{[2
]}

Do Nascimento, Jessica M. ^{[2
]}

Da Silva Sobral, Renata, V ^{[1
]}

Do Couto, Milena T. T. ^{[1
]}

Nascimento, Thiago P. ^{[2
]}

Costa, Romero M. P. B. ^{[3
]}

Porto, Ana Lucia F. ^{[2
]}

Leite, Ana Cristina L. ^{[1
]}

机构：

[1] Univ Fed Pernambuco, Dept Ciencias Farmaceut, Lab Biotecnol & Hemoderivados, Ave Artur Sa, BR-50740520 Recife, PE, Brazil

[2] Univ Fed Rural Pernambuco, Dept Morfol Anim, Lab Prod Bioativos & Tecnol, Ave Dom Manuel Medeiros, BR-52171900 Recife, PE, Brazil

[3] Univ Pernambuco, Inst Ciencias Biol, Lab Avancado Biotecnol Proteinas, Rua Arnobio Marques 310, BR-50100130 Recife, PE, Brazil

来源：

ANAIS DA ACADEMIA BRASILEIRA DE CIENCIAS | 2021年 / 93卷

关键词：

Actinomycetes; protease; fibrinoLysis; thrombolytic; SERINE-PROTEASE; EXTRACTION; ENZYME; FIBRINOGEN; MUSHROOM;

D O I：

10.1590/0001-3765202120210335

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Fibrinolytic proteases are a promising alternative in the pharmaceutical industry, they are used in the treatment of cardiovascular diseases, especially thrombosis. Microorganisms are the most interesting source of fibrinolytic proteases. The aim of this study was the production of fibrinolytic protease from Streptomyces parvulus DPUA 1573, the recovery of the protease by aqueous two-phase system and partial biochemical characterization of the enzyme. The aqueous two-phase system was performed according to a 2(4)-full factorial design using polyethylene glycol molar mass, polyethylene glycol concentration, citrate concentration and pH as independent variables. It was analyzed the effect of different ions, surfactants, inhibitors, pH and temperature on enzyme activity. The best conditions for purifying the enzyme were 17.5% polyethylene glycol 8,000, 15% Phosphate and pH 8.0, it was obtained a partition coefficient of 7.33, a yield of 87.49% and a purification factor of 2.10-fold. There was an increase in enzyme activity in the presence of Fe2+ and a decrease in the presence of beta-Mercaptoethanol, phenylmethylsukfonyl fluoride and lodoacetic acid. The optimum pH was 7.0 and the optimum temperature was 40 degrees C. The purified protease exhibited a molecular mass of 41 kDa. The fibrinolytic protease from Streptomyces parvulus proved to be a viable option for the development of a possible drug with fibrinolytic action.

引用

页数：17

共 48 条

[1] Recovery and partial purification of fibrinolytic enzymes of Auricularia polytricha (Mont.) Sacc by an aqueous two-phase system [J].