具有纤溶活性作用的海洋真菌代谢产物的分离与菌株鉴定

摘要

本文旨在分离具有纤溶活性作用的化合物和鉴定产生纤溶活性化合物的菌株FG216的种属分类.以马铃薯蔗糖培养基为种子培养基,改良查氏培养基为发酵培养基对菌株进行发酵培养,用甲醇作为提取溶剂,通过半制备型高效液相色谱从真菌FG216的1L发酵液中分离和精制了12 mg纤溶活性化合物,该纤溶活性化合物在纤溶酶原和单链尿激酶性纤溶酶原激活剂相互活化反应体系中添加l0 μg/mL活性最高.对FG216菌株rDNA的ITS基因(ITS-5.8 S rDNA)进行PCR扩增、测序,从GenBank获取相似序列,通过序列比对分析和系统发育分析表明菌株FG216与Stachybotrys longispora同源性最高.从分离的海洋微生物葡萄穗霉属菌株FG216分离得到的纤溶活性化合物具体促进纤溶酶原和单链尿激酶性纤溶酶原激活剂相互活化的作用.%To isolate fibrinolytic active compounds from cultures of marine fungi FG216 and initial identify of the strain, czapek medium was choosed as fermentation medium. The active compounds with fibrinolytic' activity measured by micro-plate reader were isolated from a culture broth and refined with marine fungi FG216 by using semi-preparative HPLC. In the reciprocal activation system of prourokinase and plasminogen.the activity was highest when the additive amount of fibrinolytic active compound was 10 μg/mL. The complete ITS sequence of FG216 strain was cloned and sequenced, and phylogenetic tree based on ITS-5.8 S rDNA sequences showed that FG216 strain has a high homology with Stachybotrys longispora on taxonomic level. Reciprocal activation of prourokinase and plasminogen was enhanced by the fibrinolytic active compound with the strain of marine fungi FG216.