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首页> 外文期刊>World Journal of Microbiology and Biotechnology >Screening of a high fibrinolytic enzyme producing strain and characterization of the fibrinolytic enzyme produced from Bacillus subtilis LD-8547
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Screening of a high fibrinolytic enzyme producing strain and characterization of the fibrinolytic enzyme produced from Bacillus subtilis LD-8547

机译：高产纤溶酶的菌株的筛选和枯草芽孢杆菌LD-8547产生的纤溶酶的表征

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摘要

A fibrinolytic enzyme producing strain Bacillus subtilis LD-8 was isolated from douchi, a traditional Chinese soybean-fermented food. After mutagenesis treatments by UV, NTG (N-methyl-N′-nitroso-N-nitroso-guanidine) and γ-radiation, a high fibrinolytic enzyme producing strain B. subtilis LD-8547 was obtained. Under optimum condition, LD-8547 was able to yield the average fibrinolytic activity of 4220 U/mL in 15 L fermenter. The strong fibrin-specific enzyme was purified from supernatant of B. subtilis LD-8547 culture broth using the combination of various steps. The optimal temperature and pH value of this fibrinolytic enzyme were 50 °C and 8.0, respectively. The molecular weight was about 30 kDa measured by SDS-PAGE. The amidolytic activity of this fibrinolytic enzyme was inhibited completely by 1 mmol/L phenylmethanesulfonyl fluoride (PMSF), but EDTA and EGTA did not affect the enzyme activity. The apparent K m and V max values were 0.521 mmol/L and 0.049 mmol/min, respectively. In vitro assays revealed that the enzyme could catalyze blood clot lysis effectively, indicating that this enzyme could be a useful thrombolytic agent.
机译：从中国传统的大豆发酵食品-豆isolated中分离出产纤溶酶的枯草芽孢杆菌LD-8。通过紫外线，NTG（N-甲基-N'-亚硝基-N-亚硝基胍）和γ-射线诱变处理后，获得了一种高产纤溶酶的枯草芽孢杆菌LD-8547。在最佳条件下，LD-8547在15 L发酵罐中的平均纤维蛋白溶解活性为4220 U / mL。使用各种步骤的组合，从枯草芽孢杆菌LD-8547培养液的上清液中纯化强纤维蛋白特异性酶。该纤溶酶的最佳温度和pH值分别为50°C和8.0。通过SDS-PAGE测量的分子量为约30kDa。该纤溶酶的酰胺分解活性被1 mmol / L苯基甲磺酰氟（PMSF）完全抑制，但EDTA和EGTA并未影响该酶的活性。表观K m 和V max 值分别为0.521 mmol / L和0.049 mmol / min。体外测定表明该酶可以有效催化血凝块溶解，表明该酶可能是有用的血栓溶解剂。

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