湛江沿海潮间带产胞外纤溶活性酶类海洋真菌的生物多样性分析

【目的】研究湛江沿海硇洲岛和徐闻珊瑚礁自然保护区潮间带产胞外纤溶酶样酶和纤溶酶原激活物海洋真菌的生物多样性,为发掘新型溶栓药物奠定基础。【方法】采用马铃薯葡萄糖琼脂(PDA)和酵母膏蛋白胨葡萄糖(YPD)培养基分离培养海洋真菌,采用真菌r DNA转录间隔区1-5.8S r DNA-转录间隔区2(ITS1-5.8S-ITS2)片段的序列分析及其系统进化树构建的方法鉴定分离培养的海洋真菌,采用脱脂牛奶马铃薯葡萄糖琼脂(SM-PDA)培养基培养法筛选产胞外蛋白酶的海洋真菌,采用海水纤维蛋白马铃薯葡萄糖琼脂(FN-PDA)培养基培养法筛选产胞外纤溶酶样酶和/或纤溶酶原激活物的海洋真菌。【结果】从湛江沿海的硇洲岛和徐闻珊瑚礁自然保护区潮间带分离、培养和鉴定了海洋真菌446株,含真菌的98个种,分布于真菌域子囊菌门(Ascomycota)和担子菌门(Basidiomycota)的6个纲、18个目、46个科、65个属;其中产胞外蛋白酶的海洋真菌有265株,61个种,分布于41个属;产胞外纤溶酶样酶的海洋真菌有67株,22个种,分布于14个属;产胞外纤溶酶原激活物的海洋真菌有84株,23种,分布于13个属;优势属为曲霉属(Aspergillus),其次为青霉属(Penicillium)。【结论】湛江沿海潮间带可分离培养的产胞外纤溶酶样酶和纤溶酶原激活物的海洋真菌物种丰富多样,是发掘新型溶栓药的丰富资源。

Biodiversity analysis of marine fungi producing extracellular fibrinolytic activity enzymes from the intertidal zone along Zhanjiang coast

Objective] To investigate the biodiversity of marine fungi producing extracellular plasmin-like enzymes and plasminogen activators from the intertidal zone of Naozhou island and Xuwen coral reef nature reserve along Zhanjiang coast, and lay the foundation for discovery of new thrombolytics of plasmin-like enzymes and plasminogen activators. Methods] Marine fungi were isolated and cultured by the method using potato dextrose agar (PDA) culture plates and yeast extract peptone dextrose (YPD) media, and identified by sequence analysis and phylogenetic tree construction of fungal rDNA internal transcribed spacer 1-5.8S rDNA-internal transcribed spacer 2 (ITS1-5.8S-ITS2) fragments. The marine fungi producing extracellular proteases were screening by the method using skim milk PDA (SM-PDA) plates, and the marine fungi producing extracellular plasmin-like enzymes and plasminogen activators were screening by the method using sea water fibrin PDA (FN-PDA) plates. Results] A total of 446 strains of marine fungi were isolated, cultured and identified from the intertidal zone of Naozhou island and Xuwen coral reef nature reserve along Zhanjiang coast, which contained 98 species distributed in 65 genera, 46 families, 18 orders, 6 classes, belonged to Ascomycota and Basidiomycota of fungus. Amongst, there were 265 strains of marine fungi producing extracellular proteases represented 61 species distributed in 41 genera, 67 strains of fungi producing extracellular plasmin-like enzymes represented 22 species distributed in 14 genera, and 84 strains of fungi producing extracellular plasminogen activators represented 23 species distributed in 13 genera. The dominant genus was Aspergillus, and then Penicillium. Conclusion] There were species biodiversity of the culturable marine fungi producing extracellular plasmin-like enzymes and plasminogen activators in the intertidal zone along Zhanjiang coast, which would be the rich resources for discovery of new thrombolytic agents.