title: Biochemical analysis of a fibrinolytic enzyme purified from Bacillus subtilis strain A1.
abstract: A fibrinolytic enzyme from Bacillus subtilis strain Al 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 Al 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 Al.
authors: Department of Biotechnology, Dong-A University, Busan, 604-714, Republic of Korea.
authors: Won Sik
authors: WS
authors: Yeo
authors: Min Jeong
authors: MJ
authors: Seo
authors: Min Jeong
authors: MJ
authors: Kim
authors: Hye Hyeon
authors: HH
authors: Lee
authors: Byoung Won
authors: BW
authors: Kang
authors: Jeong Uck
authors: JU
authors: Park
authors: Yung Hyun
authors: YH
authors: Choi
authors: Yong Kee
authors: YK
authors: Jeong
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D002918:Chymotrypsin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D005337:Fibrin / Q000378:metabolism*; D005343:Fibrinolytic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D057057:Serine Proteases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D005343:Fibrinolytic Agents; D005337:Fibrin; D057057:Serine Proteases; D002918:Chymotrypsin