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Fibrinolytic Enzyme Production from Bacilus subtilis A1-W
여원식,갈상완,정영기 한국생명과학회 1998 한국생명과학회 학술발표회 Vol.20 No.-
A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. Optimum culture conditions for the production of fibrinolytic enzyme was determined to be 1.5% soluble starch and 0.5% peptone as nutrient sources, (NH₄)₃P0₃ . 3H₂0 and MgS0₄·7H₂0 as phosphorus source and metal ions. Initial pH and temperature were pH 8.0 and 30℃ and it needed high aeration. As growing cell under the optimum culture conditions, the fibrinolyic activity increased gradually and the highest enzyme production was observed after 30 hours of cultivation at 30℃.
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Biochemical Analysis of a Fibrinolytic Enzyme Purified from Bacillus subtilis Strain A1
여원식,서민정,김민정,이혜현,강병원,박정욱,최영현,정영기 한국미생물학회 2011 The journal of microbiology Vol.49 No.3
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.
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