Production, Screening and Purification of a Fibrinolytic Enzyme (Actinokinase) from Streptomyces species
Production, Screening and Purification of a Fibrinolytic Enzyme (Actinokinase) from Streptomyces species
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Date
2024-12
Authors
Marvet Abdella Mohamed
Hassan Bashier Elamin
Motaz Nasir Hassan
Hanan Moawia Ibrahim
Journal Title
Journal ISSN
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Publisher
Napata College
Abstract
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.
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Original article