地学前缘 ›› 2008, Vol. 15 ›› Issue (6): 192-198.

• 论文 • 上一篇    下一篇

石油脱硫微生物菌株的筛选及鉴定的研究

  

  1. 1中国地质科学院 地质力学研究所, 北京 100081
    2北京大学 化学与分子工程学院, 北京 100871

  • 出版日期:2008-06-20 发布日期:2008-06-20
  • 作者简介:田晓娟（1954—），女，副研究员，主要从事化学分析和地球化学研究工作。E-mail:xjtianlxs@126.com
  • 基金资助:

    中国地质调查局项目(1212010633603)

Research on identification and screen of microbial desulfurization strains for petroleum.

  1. 1Institute of Geomechanics,Chinese Academy of Geological Sciences, Beijing 100081, China
    2College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China

  • Online:2008-06-20 Published:2008-06-20

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摘要：

从胜利油田和冀东油田采集被石油污染的土壤样品,经富集技术培养后,从中分离出21株脱硫菌,再从中筛选出一高效脱硫菌株TV9704。这一菌株不能以正十二烷、正十六烷、液体石蜡、萘和柴油为碳源和能源生长,因此不会明显地损失石油的燃烧值;但它能利用噻吩或二苯并噻吩(DBT)作为唯一硫源来生长。本实验中噻吩和DBT的浓度使用UV分光光度计测定。菌株TV9704在噻吩起始浓度为632 mmol/L的培养基中分别培养48 h和144 h后,它对噻吩的降解率分别为390％、638％;在DBT起始浓度为27 mmol/L的培养基中培养72 h,DBT降解了146 mmol/L。选择醋酸钠和甘油为碳源时,乙醇能显著提高TV9704对DBT的降解率。菌株TV9704经中国工业微生物菌种保藏管理中心(CICC)鉴定为芽孢杆菌(Bacillus sp),革兰氏阳性,专性好氧,在营养肉汁平板上形成橙黄色的圆形菌落。对菌株TV9704所作16SrDNA基因序列测定和分析显示,它与最相似种Bacillus aquimaris,Bacillus marisflavi的同源性分别为992％和982％,但它们之间的细胞形态特征及生理生化特性相差较大,因此无法鉴定到种。菌株TV9704可能是一个新种。

关键词: 生物脱硫; 菌株; 噻吩;二苯并噻吩; 筛选; 芽孢杆菌

Abstract:

Cultured with enrichment technology, a high desulfurization strain TV9704 was further selected from the 21 desulfurization bacteria separated from oilcontaminated soil samples of the Jidong Oilfield and Shengli Oilfield. The strain TV9704 does not use dodecane, hexadecane, liquid paraffin, diesel and naphthalene as carbon source and energy for its growth, therefore the oil consumption value was not obviously decreased; comparatively, thiophene or dibenzothiophene(DBT) was the only carbon source, sulfur source and energy source. In the experiment, the concentration of thiophene or DBT was measured by UV spectrophotometer. After 48 hs and 144 hs cultured with culture medium of initial concentration of 632 mmol/L, the degradation rate of thiophene by the strain TV9704 was 390% and 638% respectively. After 72 hs cultivation, the degradation of DBT by the strain TV9704 (initial concentration of 27 mmol/L) was 124 mmol/L. Using glycerol and sodium acetate as carbon source, the degradation rate of DBT was significantly increased by ethanol. Identified by China Industrial Culture Collection Center(CICC), the strain TV9704 was classified as Bacillus sp, with the characteristics of grampositive, obligate aerobic, and a circular orange colony formed on the panel covered with nutrition gravy. With the 16SrNDA gene sequencing test and analysis, the homology of the strain TV 9704 to Bacillus aquimaris and Bacillus misflav was 992% and 982%, respectively. However, their cell morphology as well as their physiological and biochemical characteristics were very different from each other. We suppose the strain TV9704 to be a fresh new genus since it is impossible to categorize it to any population.

Key words:

 Biodesulfurization; strain; thiophene; dibenzothiophene; screening; Bacillus sp

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