Issue 3

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Browsing Issue 3 by Author "Motaz Nasir Hassan"

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    Production, Screening and Purification of a Fibrinolytic Enzyme (Actinokinase) from Streptomyces species
    (Napata College, 2024-12) Marvet Abdella Mohamed; Hassan Bashier Elamin; Motaz Nasir Hassan; Hanan Moawia Ibrahim
    Aim: Cardiovascular diseases associated with thrombosis are one of the main causes of death all around the world. Urokinase, streptokinase, and tissue plasminogen activator are the major thrombolytic agents used to treat thrombosis. The fact that these agents have several side effects and expensive, has driven researchers to search for safer and more economically viable compounds for the treatment of cardiovascular diseases. Thus, the aim of this study is to evaluate the potential of Streptomyces isolated from local soil to produce fibrinolytic enzyme. Materials and methods: Three hundred total of soil samples were collected from different areas in Sudan, the isolates were identified using molecular markers. The enzyme was extracted from fermentation broth. The extract was concentrated by precipitating with (30%- 90%) ammonium sulfate salt, the precipitate fractions were obtained by dialysis membrane and Sephadex G-75 gel permeation chromatography. The purified enzyme was characterized in term of pH, temperature and time of storage. The time of complete lysis of blood clot in-vitro was also calculated. Results: The study revealed that 68% of the isolates were positive for thermophilic Streptomyces sp. (41%) of the positive isolates could product actinokinase. Identification of the isolate confirmed it is a thermophilic Streptomyces megasporus. The pH had dropped toward acidic (5.7) at the fermentation broth as an indicator of successful actinokinase production. Purification of actinokinase could produce 5.7mg/mL of total protein, with a specific activity of 101U/mg-1 , and yield of (70%). The enzyme was stable at a broad range of pH ranging from 6 to 10, and had high thermostable activity at 37oC for 6 months. SDS-PAGE showed that the molecular mass of actinokinase was approximately 35 KDa. The enzyme kinetic revealed that the Vmax and Km, were found to be 8.02 µmol /ml/min and 0.56 µmol /ml/min, respectively. In vitro fibrin degradation showed that complete clot lysis was attained within 20 minutes. Conclusion The study findings indicated that the isolates Streptomyces from local soil in Sudan were capable of producing fibrinolytic enzyme actinokinase that specifically act on fibrin and could lyse blood clot within 20 minutes.