四株碱性环境真菌铁载体活性筛选

Screening siderophore activity of four strains from alkaline environment

【目的】铁载体(Siderophore)是由微生物产生的一类低分子量金属离子螯合物。在生物医药、环境修复、健康食品等领域均具有广泛应用前景。文献调查显示,针对碱性环境真核微生物铁载体研究尚无相关报道。从中筛选高活性铁载体真菌具有重要意义。【方法】采用铬天青S显色法对分离于云南省碱性湖泊-程海和个旧大屯碱性尾矿土壤的99株真菌进行筛选;利用"分光光度法"对菌株产铁载体能力和铁载体类型进行考察;采用固相萃取(SPE)手段对菌株铁载体富集效果进行考察;根据菌体形态电镜观察和ITS基因系统发育树构建,对菌株进行生物学鉴定。【结果】通过初筛和复筛,确定菌株FEDT-866、FEDT-145、FECH-998和FECH-595均为铁载体高产菌株;所产铁载体类型主要为异羟肟酸型和羧酸型。除菌株FEDT-866外,铁载体活性产物适合采用固相萃取(SPE)方法进行富集。经生物学鉴定,菌株FEDT-866和FECH-998属于曲霉属,分别与Aspergillus tubigensis和A.nomius相似性较高;菌株FECH-595和FEDT-145属于青霉属,分别与Penicillium svalbardense和P.chrysogenum相似性较高。【结论】P.chrysogenum是1种常见的产铁载体真菌。而A.tubigensis、A.nomius及P.svalbardense菌株尚无产铁载体相关报道,可作为铁载体研究良好材料。

[Objectives] Siderophore is a low molecular iron chelate produced by microorganisms. It has broad application prospects in medicine, environmental restoration, health food and other fields. According to the literature survey so far, no siderophore was found from alkaline environment eukaryotes. Therefore, screening of fungi with high siderophore activity is of great significance. [Methods] By chromium azure S coloration, we screened 99 fungi isolated from Cheng Hai（an alkaline lakes in Yunnan province） and Datun alkaline tailings（Gejiu, Yunnan province）.By spectrophotometric detection, we investigated the strain capacity of siderophore and type of siderophore. By solid phase extraction, we investigated the siderophore enrichment effect. Based on electron microscopy morphologic observation and ITS gene phylogenetic tree construction, we identified the strain. [Results] Strains FEDT-866, FEDT-145, FECH-998 and FECH-595 were siderophore high-yield ones. Except for strain FEDT-866, the siderophore active substances were suitable for solid phase extraction（SPE）. Strains FEDT-866 and FECH-998 belong to Aspergillus and have higher similarity with Aspergillus tubigensis and A. nomius, respectively. Strains FECH-595, FEDT-145 belong to Penicillium and have higher similarity with P. svalbardense and P. chrysogenum. [Conclusion] We isolated and identified four fungi for possible siderophore production.