Novel thrombolytic protease from edible and medicinal plant Aster yomena (Kitam.) Honda with anticoagulant activity: Purification and partial characterization

• Published in: Journal of bioscience and bioengineering Vol. 118; no. 4; pp. 372 - 377
• Main Authors: CHOI, Jun-Hui, KIM, Dae-Won, PARK, Se-Eun, CHOI, Bong-Suk, SAPKOTA, Kumar, SEUNG KIM, KIM, Sung-Jun
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier 01.10.2014
• Subjects: Animals; Aster Plant - chemistry; Aster Plant - enzymology; Biological and medical sciences; Biotechnology; Blood Coagulation Tests; Cations, Divalent; Collagen; Edetic Acid - chemistry; Egtazic Acid - chemistry; Endopeptidases - isolation & purification; Endopeptidases - metabolism; Endopeptidases - pharmacology; Fibrin - chemistry; Fibrin - metabolism; Fibrinogen - chemistry; Fibrinogen - metabolism; Fibrinolysis - drug effects; Fibrinolytic Agents - isolation & purification; Fibrinolytic Agents - metabolism; Fibrinolytic Agents - pharmacology; Fundamental and applied biological sciences. Psychology; Hot Temperature; Kinetics; Male; Mice; Mice, Inbred ICR; Molecular Weight; Plant Leaves - chemistry; Plant Proteins - isolation & purification; Plant Proteins - metabolism; Plant Proteins - pharmacology; Plants, Medicinal; Pulmonary Embolism - blood; Pulmonary Embolism - chemically induced; Pulmonary Embolism - drug therapy; Zinc - chemistry; Purification; Enzyme; Aster; Anticoagulant; Compositae; Fibrinolysis; Medicinal plant; Characterization; Peptidases; Dicotyledones; Angiospermae; Antithrombotic; Aster yomena (Kitam.) Honda; Hydrolases; Spermatophyta
• ISSN: 1389-1723; 1347-4421
• DOI: 10.1016/j.jbiosc.2014.03.004

A thrombolytic protease named kitamase possessing anticoagulant property was purified from edible and medicinal plant Aster yomena (Kitam.) Honda. Kitamase showed a molecular weight of 50 kDa by SDS-PAGE and displayed a strong fibrin zymogram lysis band corresponding to the similar molecular mass. The enzyme was active at high temperatures (50°C). The fibrinolytic activity of kitamase was strongly inhibited by EDTA, EGTA, TPCK and PMSF, inhibited by Zn(2+). The Km and Vmax values for substrate S-2251 were determined as 4.31 mM and 23.81 mM/mg respectively. It dissolved fibrin clot directly and specifically cleaved the α, Aα and γ-γ chains of fibrin and fibrinogen. In addition, kitamase delayed the coagulation time and increased activated partial thromboplastin time and prothrombin time. Kitamase exerted a significant protective effect against collagen and epinephrine induced pulmonary thromboembolism in mice. These results suggest that kitamase may have the property of metallo-protease like enzyme, novel fibrino(geno)lytic enzyme and a potential to be a therapeutic agent for thrombosis.