Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7615025 | Journal of Chromatography B | 2018 | 20 Pages |
Abstract
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 (1637.45â¯Â±â¯15â¯mgâ¯Lâ1) and productivity (181.93â¯mgâ¯Lâ1â¯dayâ1) 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 1834.6â¯Uâ¯mgâ1 and 226.86â¯mm2 by spectrophotometry and fibrin plate assays, respectively. SDS-PAGE results exhibited a protein band of about 45â¯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 90â¯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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Authors
Páblo Eugênio da Costa e Silva, Rafaela Cavalcante de Barros, Wendell Wagner Campos Albuquerque, Romero Marcos Pedrosa Brandão, Raquel Pedrosa Bezerra, Ana Lúcia Figueiredo Porto,