【目的】从油页岩环境中筛选可降解油页岩干酪根的产生物表面活性剂菌株。【方法】从抚顺油页岩矿废水样品中用血平板法初筛,排油圈法、乳化法和表面张力法复筛,获得产生物表面活性剂菌株。对目标菌株进行生理生化鉴定、16S r RNA基因序列和系统发育分析,用薄层色谱鉴定其发酵液表面活性成分,优化产表面活性剂的培养条件,初步考察其对油页岩干酪根的降解能力。【结果】筛选到一株产糖脂表面活性剂菌株B-1,初步鉴定为Pseudomonas sp.,该菌株有良好的排油和乳化能力以及较低的表面张力,可利用烷烃、不饱和脂肪酸和糖类作为碳源。在30-34°C范围内添加0.3%Na Cl的葡萄糖培养基(p H 7.0)中该菌生长旺盛,发酵液表面张力最低为27 m N/m。菌株B-1在添加一定量葡萄糖的无机盐培养基中作用30 d后对干酪根的降解率为2.85%,高于不添加葡萄糖无机盐培养基对照组的降解率(1.04%)。【结论】菌株B-1是一株性能良好的产糖脂表面活性剂细菌,有降解干酪根的潜力。
As an insufficiently utilized energy resource,oil shale is conducive to the formation of characteristic microbial communities due to its special geological origins.However,little is known about fungal diversity in oil shale.Polymerase chain reaction cloning was used to construct the fungal ribosomal deoxyribonucleic acid internal transcribed spacer(r DNA ITS)clone libraries of Huadian Mine in Jilin Province,Maoming Mine in Guangdong Province,and Fushun Mine in Liaoning Province.Pure culture and molecular identification were applied for the isolation of cultivable fungi in fresh oil shale of each mine.Results of clone libraries indicated that each mine had over 50% Ascomycota(58.4%–98.9%)and 1.1%–13.5%unidentified fungi.Fushun Mine and Huadian Mine had 5.9% and 28.1% Basidiomycota,respectively.Huadian Mine showed the highest fungal diversity,followed by Fushun Mine and Maoming Mine.Jaccard indexes showed that the similarities between any two of three fungal communities at the genus level were very low,indicating that fungi in each mine developed independently during the long geological adaptation and formed a community composition fitting the environment.In the fresh oil-shale samples of the three mines,cultivable fungal phyla were consistent with the results of clone libraries.Fifteen genera and several unidentified fungi were identified as Ascomycota and Basidiomycota using pure culture.Penicillium was the only genus found in all three mines.These findings contributed to gaining a clear understanding of current fungal resources in major oil-shale mines in China and provided useful information for relevant studies on isolation of indigenous fungi carrying functional genes from oil shale.
