| Title |
Purification and Biochemical Characterization of a 17 kDa Fibrinolytic Enzyme from Schizophyllum commune |
| Author |
In Suk Park1, Jeong Uck Park2, Min Jeong Seo3, Min Jeong Kim1, Hye Hyeon Lee1, Sung Ryeal Kim1, Byoung Won Kang2, Yung Hyun Choi4, Woo Hong Joo5, and Yong Kee Jeong1,2,3* |
| Address |
1Department of Biotechnology, Dong-A University, Busan 604-714, Republic of Korea, 2Medi-Farm Industrialization Research Center, Dong-A University, Busan 604-714, Republic of Korea, 3Department of Medical Bioscience, Dong-A University, Busan 604-714, Republic of Korea, 4Department of Biochemistry, College of Oriental Medicine, Dong-Eui University, Busan 614-050, Republic of Korea, 5Department of Biology, Changwon National University, Kyungnam 641-713, Republic of Korea |
| Bibliography |
Journal of Microbiology, 48(6),836-841, 2010,
|
| DOI |
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| Key Words |
fibrinolytic enzyme, fibrin-zymography, N-terminal amino acid sequence, mushroom, S. commune |
| Abstract |
A fibrinolytic enzyme of the mushroom, Schizophyllum commune was purified with chromatographic methods, including a DEAE-Sephadex A-50 ion-exchange column and gel filtrations with Sephadex G-75 and Sephadex G-50 columns. The analysis of fibrin-zymography and SDS-PAGE showed that the enzyme was a monomeric subunit that was estimated to be approximately 17 kDa in size. The fibrinolytic activity of the enzyme in plasminogen-rich and plasminogen-free fibrin plates was 1.25 and 0.44 U/ml, respectively. The N-terminal amino acid sequence of the purified enzyme was identified as HYNIXNSWSSFID, which was highly distinguished from known fibrinolytic enzymes. The relative activity of the purified enzyme with an addition of 5 mM EDTA, Phosphoramidon, and Bestatin was about 76, 64, and 52%, respectively, indicating that it is a metalloprotease. The optimum temperature for the purified enzyme was approximately 45°C, and over 87% of the enzymatic activity was maintained as a stable state in a pH range from 4.0 to 6.0. Therefore, our results
suggest that the potential thrombolytic agent from S. commune is a unique type of fibrinolytic enzyme. |
