生物技术通报 ›› 2016, Vol. 32 ›› Issue (8): 184-193.doi: 10.13560/j.cnki.biotech.bull.1985.2016.08.027

• 研究报告 • 上一篇    下一篇

海洋石油降解菌的筛选及复合菌系的构建

吴秉奇,刘淑杰,陈福明,周楚莹   

  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,深圳 518057
  • 修回日期:2015-10-22 出版日期:2016-08-25 发布日期:2016-08-25
  • 作者简介:吴秉奇,男,硕士,研究方向:环境微生物;E-mail:wubingqi@163.com
  • 基金资助:
    深圳市科技计划项目(JCYJ20120817102941279)

Screening of Marine Crude Oil-degrading Bacteria and Construction of Microbial Consortium

WU Bing-qi, LIU Shu-jie, CHEN Fu-ming, ZHOU Chu-ying   

  1. Research Institute of Tsinghua University in Shenzhen,Shenzhen Environmental Microbiology Resources Development and Application Engineering Laboratory,Shenzhen 518057
  • Revised:2015-10-22 Published:2016-08-25 Online:2016-08-25

摘要: 为采用生物法治理海洋石油污染,以原油为唯一碳源,从深圳海域5个采样点取样,通过富集、涂布平板分离高效石油降解菌,并以复配、正交等方式构建石油降解复合菌系;通过生理生化实验和16S rRNA 基因序列分析对菌株进行鉴定;采用单因素实验对复合菌系降解石油的条件进行优化,并使用气相色谱-质谱法(GC-MS)研究其对石油的降解特性。结果显示,共分离得到22株高效石油降解菌,对石油的降解率为34.5%-52.2%;由S1-30、S1-38和S2-13构建了复合菌系SQ1,对石油降解率可达68.3%,3株菌分别鉴定为棒杆菌(Corynebacterium sp.)、迪茨氏菌(Dietzia sp.)和拉布伦茨氏菌(Labrenzia sp.);经优化,SQ1在最适条件下(30℃、pH7.6、石油浓度20 g/L),11 d内对石油的去除率高达73.5%;GC-MS结果表明,复合菌系SQ1对石油总烷烃的去除率为91.7%,对较难降解的C21-C35烷烃组分的降解率接近100%。研究表明,复合菌系SQ1在海洋石油污染的生物修复中具有较强的应用潜力。

关键词: 石油降解菌, 复合菌系, 降解率, 鉴定, 条件优化

Abstract: For the purpose of controlling marine oil contamination by biological treatment technology,using crude oil acting as sole carbon source and enrichment and spread plate method,high-performance oil-grading bacteria were isolated from five sampling points in the sea near Shenzhen,and bacterial consortium was constructed by mixing and orthogonal experiments. Physiological and biochemical experiments and 16S rRNA gene sequence analysis were used to identify the strains. Single-factor experiment was employed to optimize the conditions of oil biodegradation by the consortium,and gas chromatography and mass spectrum(GC-MS)were utilized to analyze its biodegradation characteristics. The results showed that 22 strains of high-performance oil-degrading bacteria were isolated,and the degrading rates varied from 34.5% to 52.2%. The degrading rate by microbial consortium SQ1 composed of S1-30,S1-38,and S2-13 strains reached 68.3%. These three strains were identified as Corynebacterium sp.,Dietzia sp. and Labrenzia sp. SQ1 was able to degrade the oil by 73.5% in 11 days under optimized conditions,referring to 30℃,pH7.6,oil concentration 20 g/L. The GC-MS results showed that consortium SQ1 was able to degrade the total alkane by 91.7%,and the more refractory C21-C35 by nearly 100%. The study shows that consortium SQ1 has great application potential of bioremediation for marine oil contamination.

Key words: oil-degrading bacteria, microbial consortium, degradation rate, identification, condition optimization