| Title | A Fibrinolytic Enzyme from the Medicinal Mushroom Cordyceps militaris |
|---|---|
| Author | Jae-Sung Kim1, Kumar Sapkota1, Se-Eun Park1, Bong-Suk Choi1, Seung Kim1, Nguyen Thi Hiep1, Chun-Sung Kim1,2, Han-Seok Choi3, Myung-Kon Kim4, Hong-Sung Chun1,5, Yeal Park1, and Sung-Jun Kim1 |
| Address | 1Department of Biotechnology, BK 21Research Team for Protein Activity Control Chosun University, Gwangju 501-759, Republic of Korea, 2Department of Pharmacology, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA, 3Department of Food Science and Technology, Chonbuk National University, Jeonju 561-756, Republic of Korea, 4Department of Industrial Crop Production and Processing, Iksan National college, Iksan 570-752, Republic of Korea, 5Research Center for Proteineous Materials, Chosun University, Gwangju 501-759, Republic of Korea |
| Bibliography | Journal of Microbiology, 44(6),622-631, 2006, |
| DOI | |
| Key Words | Cordyceps militaris, Fibrinolytic enzyme, Serine protease, Subtilisin, Fibrinolysis, Thrombus |
| Abstract | In this study we purified a fibrinolytic enzyme from Cordyceps militaris using a combination of ion-exchange chromatography on a DEAE Sephadex A-50 column, gel filtration chromatography on a Sephadex G-75 column, and FPLC on a HiLoad 16/60 Superdex 75 column. This purification protocol resulted in a 191.8-fold purification of the enzyme and a final yield of 12.9%. The molecular mass of the purified enzyme was estimated to be 52 kDa by SDS-PAGE, fibrin-zymography, and gel filtration chromatography. The first 19 amino acid residues of the N-terminal sequence were ALTTQSNV THGLATISLRQ, which is similar to the subtilisin-like serine protease PR1J from Metarhizium anisopliae var. anisopliase. This enzyme is a neutral protease with an optimal reaction pH and temperature of 7.4 and 37°C, respectively. Results for the fibrinolysis pattern showed that the enzyme rapidly hydrolyzed the fibrin α-chain followed by the γ-γ chains. It also hydrolyzed the β-chain, but more slowly. The Aα, Bβ, and γ chains of fibrinogen were also cleaved very rapidly. We found that enzyme activity was inhibited by Cu2+ and Co2+, but enhanced by the additions of Ca2+ and Mg2+ ions. Furthermore, fibrinolytic enzyme activity was potently inhibited by PMSF and APMSF. This enzyme exhibited a high specificity for the chymotrypsin substrate S-2586 indicating it’s a chymotrypsin-like serine protease. The data we present suggest that the fibrinolytic enzyme derived from the edible and medicinal mushroom Cordyceps militaris has fibrin binding activity, which allows for the local activation of the fibrin degradation pathway. |
| Download PDF | JM44(6)-06.pdf |