近海柴油降解菌群的构建及其对柴油的降解特性  

Construction of diesel fuel-degrading bacterial consortium from coastal waters and its characteristics on degradation of diesel fuel

作　　者：周楚莹[1,2] 吴秉奇[1,3] 刘淑杰[1,3] 陈福明[1,3] 杨小龙[1,3] 刘国光[2] 

机构地区：[1]深圳清华大学研究院深圳环境微生物资源开发与应用工程实验室,广东深圳518057 [2]广东工业大学环境科学与工程学院,广东广州330063 [3]深圳市清研环境科技有限公司,广东深圳518057

出　　处：《微生物学通报》2015年第12期2308-2320,共13页Microbiology China

基　　金：深圳市科技计划项目(No.JCYJ20120817102941279)

摘　　要：【目的】实施近海柴油污染的生物治理。【方法】以柴油为唯一碳源,从深圳港口海域富集筛选柴油降解菌;采用复配、正交试验等方式构建混合菌群;通过单因素试验研究环境因素对菌群降解柴油的影响;使用气相色谱-氢火焰检测器(GC-FID)分析降解前后柴油各组分的变化;通过生理生化试验和16S r RNA基因序列分析对菌株进行鉴定。【结果】获得了16株柴油降解菌,7 d内对柴油的降解率最高达40.8%;选择菌株C1-8、C2-10、C3-13构建了混合菌群CQ1,投加量分别为0.5%、2.0%和1.0%,CQ1对柴油去除率比单菌提高了10%以上;CQ1的最适环境条件为:温度30°C、p H 7.6、摇床转速220 r/min、柴油浓度20 g/L,优化后9 d内对柴油去除率达60%以上;GC-FID结果显示,菌群CQ1可降解大部分C11-C27的正构烷烃,对C21-C27的烷烃降解可达100%。经鉴定,菌株C1-8、C2-10和C3-13分别为微杆菌(Microbacterium sp.)、剑菌(Ensifer sp.)和变异棒杆菌(Corynebacterium variabile)。【结论】CQ1在近海柴油污染的生物修复中具有良好的应用前景。[Objective] For the purpose of control marine diesel pollution by biological treatment technology. [Methods] Diesel act as sole carbon source, using enrichment and screening methods to select diesel fuel-degrading bacteria from Shenzhen harbor area; bacterial consortium is constructed bymixing and orthogonal experiments; single-factor experimental is used to study the effect of environmental factors on diesel biodegradation by the consortium; using chromatography-flame ionization detection （GC-FID） to analyze the changes of diesel components before and after degradation, physiological, biochemical experiment and 16S rRNA gene sequence analysis are used to identify the strains. [Results] Sixteen diesel fuel-degrading bacteria were isolated and the highest degrading rates were up to 40.8% in 7 days. Bacteria consortium CQ1 was composed of C1-8, C2-10 and C3-13 strains, and the dosage was 0.5%, 2.0% and 1.0% respectively. Compared to the single strain, the degrading rate of CQ 1 was increased by more than 10%. The best degrading effects would be up to 60% in 9 days under the optimum conditions that 30 ℃, pH 7.6, shaking speed 220 r/min and diesel concentration 20 g/L. The result of GC-FID showed that most n-alkanes Cll-C27 can be degraded by bacteria consortium CQ1 and the degradation rate of C21-C27 were up to 100%. In addition, the results showed that C1-8 belonged to Microbacterium sp., C2-10 belonged to Ensifer sp., C3-13 belonged to Corynebacterium variabile. [Conclusion] CQ1 has bright application prospects of bioremediation for offshore diesel pollution.

关 键 词：柴油降解菌 菌群 降解率 环境因素 鉴定 

分 类 号：X172[环境科学与工程—环境科学] X55