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Cross-linked Enzyme Aggregates of Fibrinolytic Protease BC1 Immobilized on Magnetic Chitosan Nanoparticles (CLEAs-Fib-mChi): Synthesis, Purification, and Characterization

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Abstract

Bacterial fibrinolytic proteases achieved more attention in the prevention and treatment of cardiovascular diseases, so purification, characterization, and activity enhancement are of prime importance. In this study, a fibrinolytic serine metalloprotease was purified from the culture supernatant from Bacillus sp. BC1. It was purified to homogeneity by a two-step procedure with a 24-fold increase in specific activity and a 33.1% yield. It showed 28 kDa molecular weight, while its optimal pH and temperature were obtained 8 and 50–60 °C. The cross-link enzyme aggregates of this fibrinolytic BC1 successfully immobilized on magnetic chitosan nanoparticles. A 52% activity enhancement was obtained by immobilized enzyme at pH 6.0, compared to free protease. Km values of the free and immobilized proteases were obtained about 0.638 and 0.61 mg/ml, respectively. The free and immobilized enzymes did not show any activity concerning transferrin, γ-globulins, and hemoglobin, as blood plasma proteins. The in vitro blood clot lysis test of the free and immobilized proteases showed a maximum of 42 and 50% clot lysis, which was comparatively higher than that revealed by streptokinase and heparin at the same condition. These results indicated that the free and immobilized proteases have the potential to be effective fibrinolytic agents.

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The authors express their gratitude to the Research Council of the Shahid Bahonar University of Kerman (Iran) and Iran National Science Foundation (INSF) for their financial support during the course of this project.

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  1. Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran

    Shima Khankari, Arastoo Badoei-dalfard & Zahra Karami

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Correspondence to Arastoo Badoei-dalfard.

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Khankari, S., Badoei-dalfard, A. & Karami, Z. Cross-linked Enzyme Aggregates of Fibrinolytic Protease BC1 Immobilized on Magnetic Chitosan Nanoparticles (CLEAs-Fib-mChi): Synthesis, Purification, and Characterization. Appl Biochem Biotechnol 193, 2004–2027 (2021). https://doi.org/10.1007/s12010-021-03494-z

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