In vitro thrombolytic activity of a purified fibrinolytic enzyme from Chlorella vulgaris
2018
Silva, Páblo Eugênio da Costa e | Barros, Rafaela Cavalcante de | Albuquerque, Wendell Wagner Campos | Brandão, Romero Marcos Pedrosa | Bezerra, Raquel Pedrosa | Porto, Ana Lúcia Figueiredo
A fibrinolytic enzyme was produced by microalga Chlorella vulgaris cultivated in autotrophic and mixotrophic conditions added corn steep liquor, purified by a single chromatographic step, then biochemical characterization and in vitro thrombolytic activity was performed. Maximum cell concentration (1637.45 ± 15 mg L−1) and productivity (181.93 mg L−1 day−1) was obtained in mixotrophic culture using 1% corn steep liquor. Enzyme-extracted microalgal biomass was purified by acetone precipitation and DEAE Sephadex anion exchange chromatography up to 2 fold with recovery of 4.0%. After purification, fibrinolytic activity was 1834.6 U mg−1 and 226.86 mm2 by spectrophotometry and fibrin plate assays, respectively. SDS-PAGE results exhibited a protein band of about 45 kDa and fibrinolytic band was detected by fibrin zymography. Enzyme activity was enhanced in the presence of Fe2+ and inhibited by phenylmethane sulfonyl fluoride (PMSF) and ethylenediamine tetracetic acid (EDTA), which suggest it to be a metal-dependent serine protease. The extract also showed a red blood cell lysis <4% and in vitro thrombolytic activity of 25.6% in 90 min of reaction. These results indicate that the fibrinolytic enzyme from C. vulgaris may have potential applications in the prevention and treatment of thrombosis.
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