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,
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.