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Updated: Oct 10, 2025
Author Spotlight: Affinity Purification of a Fibrinolytic Enzyme from Sipunculus nudus
Published on: June 2, 2023
Purification and characterization of fibrinolytic protease from Streptomyces parvulus by polyethylene
Viviane N S Alencar1, Maria Clara DO Nascimento2, Julyanne V Dos Santos Ferreira3
1Laboratório de Biotecnologia e Hemoderivados, Departamento de Ciências Farmacêuticas, Universidade Federal de Pernambuco, Avenida Artur de Sá, 50740-520 Recife, PE, Brazil.
Abstract:
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 24-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 57.49% 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, phenylmethylsulfonyl fluoride and Iodoacetic acid. The optimum pH was 7.0 and the optimum temperature was 40 ºC. 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.
Insights
Researchers produced a fibrinolytic protease from Streptomyces parvulus for potential pharmaceutical use. This enzyme aids in treating thrombosis and cardiovascular diseases, offering a promising drug development avenue.
Area of Science:
- Microbiology
- Biochemistry
- Enzymology
Background:
- Fibrinolytic proteases are crucial for treating cardiovascular diseases, particularly thrombosis.
- Microorganisms represent a significant source for discovering novel fibrinolytic proteases.
Purpose of the Study:
- To produce fibrinolytic protease from Streptomyces parvulus DPUA 1573.
- To recover the protease using an aqueous two-phase system.
- To perform partial biochemical characterization of the enzyme.
Main Methods:
- A 24-full factorial design was employed for optimizing the aqueous two-phase system.
- Independent variables included polyethylene glycol molar mass, concentration, citrate concentration, and pH.
- Enzyme activity was assessed by analyzing the effects of ions, surfactants, inhibitors, pH, and temperature.
Main Results:
- Optimal purification conditions involved 17.5% polyethylene glycol 8,000, 15% Phosphate, and pH 8.0, yielding a partition coefficient of 7.33, 57.49% yield, and a 2.10-fold purification factor.
- Enzyme activity increased with Fe2+ and decreased with $\beta$-Mercaptoethanol, phenylmethylsulfonyl fluoride, and Iodoacetic acid.
- The purified protease showed optimal activity at pH 7.0 and 40 ºC, with a molecular mass of 41 kDa.
Conclusions:
- Streptomyces parvulus is a viable source for producing fibrinolytic proteases.
- The characterized protease demonstrates potential for developing new therapeutic agents against thrombosis.
