| Title |
Biochemical Analysis of a Fibrinolytic Enzyme Purified from Bacillus subtilis Strain A1 |
| Author |
Won Sik Yeo1, Min Jeong Seo2, Min Jeong Kim1, Hye Hyeon Lee1, Byoung Won Kang2, Jeong Uck Park2, Yung Hyun Choi3, and Yong Kee Jeong1,2* |
| Address |
1Department of Biotechnology, Dong-A University, Busan 604-714, Republic of Korea, 2Medi-Farm Industrialization Research Center, Busan 604-714, Republic of Korea, 3Department of Biochemistry, College of Oriental Medicine, Dong-Eui University, Busan 614-050, Republic of Korea |
| Bibliography |
Journal of Microbiology, 49(3),376-380, 2011,
|
| DOI |
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| Key Words |
amidolytic activity, Bacillus subtilis strain A1, fibrinolytic enzyme, plasminogen activator, N-terminal amino acid sequence |
| Abstract |
A fibrinolytic enzyme from Bacillus subtilis strain A1 was purified by chromatographic methods, including DEAE Sephadex A-50 column chromatography and Sephadex G-50 column gel filtration. The purified enzyme consisted of a monomeric subunit and was estimated to be approximately 28 kDa in size by SDS-PAGE. The specific activity of the fibrinolytic enzyme was 1632-fold higher than that of the crude enzyme extract. The fibrinolytic activity of the purified enzyme was approximately 0.62 and 1.33 U/ml in plasminogen-free and plasminogen-rich fibrin plates, respectively. Protease inhibitors PMSF, DIFP, chymostatin, and TPCK reduced the fibrinolytic activity of the enzyme to 13.7, 35.7, 15.7, and 23.3%, respectively. This result suggests that the enzyme purified from B. subtilis strain A1 was a chymotrypsin-like serine protease. In addition, the optimum temperature and pH range of the fibrinolytic enzyme were 50°C and 6.0-10.0, respectively. The N-terminal amino acid sequence of the purified enzyme was identified as Q-T-G-G-S-I-I-D-P-I-N-G-Y-N, which was highly distinguished from other known fibrinolytic enzymes. Thus, these results suggest a fibrinolytic enzyme as a novel thrombolytic agent from B. subtilis strain A1. |
