| 米阳,董超,侯正欣,等.海洋链霉菌发酵纤溶酶的分离纯化和酶学性质研究[J].中国海洋药物,2016,35(3):43-48. |
| 海洋链霉菌发酵纤溶酶的分离纯化和酶学性质研究 |
| Purification and characterization of the fibrinolytic enzyme from a marine Streptomyces strain |
| 投稿时间：2015-09-18 修订日期：2015-10-20 |
| DOI： |
| 中文关键词: 海洋链霉菌 纤溶酶 分离纯化 酶学性质 |
| English Keywords:marine Streptomyces fibrinolytic enzyme purification characterization |
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| 中文摘要: |
| 目的 从筛选的1株海洋链霉菌MY0504发酵液中,分离得到1种新型纤溶酶并对其部分酶学性质进行初步研究。方法 采用高速离心、盐析、Sephadex G-75 凝胶过滤层析对纤溶酶进行分离纯化；采用纤维蛋白平板法测定纤溶活性, SDS－PAGE 电泳测定分子量,小鼠急毒实验检测安全性,并考察温度、pH、金属离子和抑制剂对酶活性的影响。结果 从发酵液提取纤溶酶的纯化倍数为7.15倍,酶活力回收率为32%,其分子量约为14kD,安全无毒。该酶在47℃以下稳定,适宜pH为7.0~9.0,最适pH值为8.0,Cu2+对该纤溶酶抑制作用显著,Ca2+有一定的促进作用。Aprotinin 强烈抑制纤溶酶活性,PMSF(苯甲基磺酰氟)可以完全抑制该纤溶酶活性,初步推测该酶是1种丝氨酸蛋白酶。结论 从海洋链霉菌MY0504发酵液中获得1种小分子量纤溶酶,为开发新的溶栓剂提供了新的选择和理论依据。 |
| English Summary: |
| Objective A novel fibrinolytic enzyme was isolated from the fermentation broth of marine Streptomyces sp. strain MY0504,which was screened from the sea water, and its partial enzymatic properties was studied preliminarily. Methods The fibrinolytic enzyme was purified by centrifugation, ammonium sulfate fractional precipitation and Sephadex G-75 gel filtration chromatography. The fibrinolytic activity was determined by fibrin plate method. The purity and molecular weight were estimated by SDS-PAGE. The safety was detected by acute toxicity test in mice. The effects of temperature, pH, metal ions and inhibitors on the fibrinolytic activity were studied. Results A fibrinolytic enzyme was purified with a purification fold of 7.15 and an activity recovery of 32%. The molecular weight of the enzyme was 14 kD, and the fibrinolytic enzyme was safe and nontoxic. The purified fibrinolytic enzyme of the strain was stable below 47℃ and pH7.0~9.0.The optimal pH was 8.0. A certain concentration of Cu2+ strongly inhibited the enzyme, and the fibrinolytic activity was slightly enhanced by the ions of Ca2+. The fibrinolytic activity was strongly inhibited by PMSF(Phenylmethylsulfonyl fluoride)and Aprotinin, suggesting that the enzyme belonged to serine protease group. Conclusion A novel fibrinolytic enzyme with low molecular weight was obtained, which might provide a new choice for thrombolytic drugs. |
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