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Purification and partial characterization of a fibrinolytic enzyme from the fruiting body of the medicinal and edible mushroom Pleurotus ferulae.
- Source :
-
Preparative biochemistry & biotechnology [Prep Biochem Biotechnol] 2017 Jul 03; Vol. 47 (6), pp. 539-546. Date of Electronic Publication: 2016 May 02. - Publication Year :
- 2017
-
Abstract
- A fibrinolytic metalloprotease with in vitro fibrinolytic effects was purified from the edible mushroom Pleurotus ferulae using several chromatography steps including anion and ion exchange, gel filtration, and fast protein liquid chromatography columns. The molecular mass of the enzyme was estimated to be 20.0 kDa, as determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fibrin zymography. The protease was active at 50°C, and pH 4.0, 5.0, and 8.0. The fibrinolytic activity of the enzyme was inhibited by ethyleneglycol-bis-(2-aminoethyl)-N,N,N',N' tetraacetic acid and strongly inhibited by two metal ions, Cu and Mg. In vitro assays evaluating fibrinolytic activity on a fibrin plate, fibrin turbidity, and thrombolytic activity on fibrin clots using human fibrinogen and human thrombin revealed that the enzyme could hydrolyze fibrin polymers directly and inhibit the formation of fibrin clots. In activated partial thromboplastin time (APTT) and prothrombin time assays, the enzyme strongly prolonged the APTT, which detects an activity of intrinsic and common pathways. The enzyme showed strong in vivo protective effect against mortality/paralysis from epinephrine plus collagen-induced acute thromboembolism in in vivo model. Our findings suggest that the enzyme may have a potential for treatment and prevention of thrombosis-relative diseases.
- Subjects :
- Animals
Blood Coagulation drug effects
Fibrinolytic Agents metabolism
Fibrinolytic Agents therapeutic use
Fruiting Bodies, Fungal metabolism
Humans
Male
Metalloproteases metabolism
Metalloproteases therapeutic use
Mice, Inbred ICR
Pleurotus metabolism
Thromboembolism blood
Thromboembolism drug therapy
Fibrinolytic Agents isolation & purification
Fruiting Bodies, Fungal enzymology
Metalloproteases isolation & purification
Pleurotus enzymology
Subjects
Details
- Language :
- English
- ISSN :
- 1532-2297
- Volume :
- 47
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Preparative biochemistry & biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 27136080
- Full Text :
- https://doi.org/10.1080/10826068.2016.1181083