外源和内源微生物在营养剂激活过程中丰度变化及对原油作用研究

生物技术通报 ›› 2013, Vol. 0 ›› Issue (11): 164-169.

外源和内源微生物在营养剂激活过程中丰度变化及对原油作用研究

王大威^1，2, 张健^1，2, 马挺³, 齐义彬³

1.海洋石油高效开发国家重点实验室，北京 100027；2.中海油研究总院，北京 100027；3.南开大学生命科学学院分子微生物学与技术教育部重点实验室，天津 300071

收稿日期:2013-05-15 出版日期:2013-11-14 发布日期:2013-11-14
作者简介:王大威，博士，工程师，研究方向：微生物采油等三次采油技术研究；E-mail：wangdw3@cnooc.com.cn
基金资助:
国家科技重大专项(2011ZX05024-004)

Study of Changes in Abundance and Degradation Crude Oil Capacity of Endogenous Bacteria and Andexogenous Bacteria Activation in Oilfield Under Injecting Activation Agent Conditions

Wang Dawei^1，2, Zhang Jian^1，2, Ma Ting³, Qi Yibin³

(1. State Key Laboratory of Offshore Oil Exploitation，Beijing 100027；2. China National Offshore Oil Corporation(CNOOC)Research Institute，Beijing 100027；3. Key Laboratory of Molecular Microbiology and Technology，Ministry of Education，College of Life Science，Nankai University, Tianjin 300071)

Received:2013-05-15 Published:2013-11-14 Online:2013-11-14

摘要/Abstract

摘要： 旨在研究在油藏环境中，营养剂激活条件下内、外源微生物相互作用过程中的细菌群落和结构的变化，及其对微生物降解原油能力的影响。利用中海油NB35-2油田A18、B16井产出液油水样，在地层条件下投放营养剂及外源菌进行富集培养后，定时取样，样品用变性梯度凝胶电泳(DGGE)技术分析其丰度结构变化；同时结合原油黏度测定、四组分分析，分析营养剂激活过程中外源、内源微生物对原油作用变化，研究营养剂的加入对激活原油降解微生物的作用。通过对内源和外源微生物激活前后DGGE条带的数量和亮度变化分析表明，油藏环境中的细菌在种类和数量上并不丰富，激活剂的加入改变了菌群原有的贫营养环境，从而使一些因营养缺乏生长受抑制细菌得以大量繁殖，菌群结构发生变化。结果显示，单独添加外源菌种不能有效激活内源菌种，加入营养剂后，内源菌种的数量和种类明显增加，同时外源菌和内源菌复配后对原油的降解作用也明显增强。

关键词: 内源细菌群落, 选择性激活, 变性梯度凝胶电泳, 稠油降解能力

Abstract: It was study the structure change of exogenous and indigenous microbial population during activation of stratal microflora under injecting activation agent conditions, and the effect on the microbial degradation crude oil capacity. The water sample was from A18、B16 well produced water in NB35-2 Oilfield. The bacteria in the water sample under injecting activation agent conditions was activated , and samples at different time intervals were collected. These samples were analyzed by denature gradient gel electrophoresis(DGGE), and the effect of addition of activation agent on microbial degradation crude oil capacity were studied through determination of viscosity and four fractions contents analysis of crude oil. Based on the analysis of the number and lightness of DGGE bands, we studied the change of the microbial community diversity activation of stratal bacteria. The species number and total quantity of dominant microbial population were not abundant under oil reservoir conditions. After injecting activation agent, some bacteria grew and reproduced quickly along with the enhancement of nutrition condition, and the structure of dominant microbial population changed. Exogenous bacteria alone can not effectively activate endogenous bacteria, and after adding nutrients, the species number and total quantity of endogenous bacteria significantly increased, while the performance of degradation to crude oil of exogenous bacteria and endogenous bacteria was significantly enhanced.

Key words: Endogenous bacterial community, Selective activation, Denature gradient gel electrophoresis, Degradation crude oil capacity

王大威, 张健, 马挺, 齐义彬. 外源和内源微生物在营养剂激活过程中丰度变化及对原油作用研究[J]. 生物技术通报, 2013, 0(11): 164-169.

Wang Dawei, Zhang Jian, Ma Ting, Qi Yibin. Study of Changes in Abundance and Degradation Crude Oil Capacity of Endogenous Bacteria and Andexogenous Bacteria Activation in Oilfield Under Injecting Activation Agent Conditions[J]. Biotechnology Bulletin, 2013, 0(11): 164-169.

参考文献

[1] 程海鹰, 王修林, 徐登霆, 等.内源微生物提高采收率实验研究[J].石油勘探与开发, 2006, 33(1)：91-95.
[2] 张雪梅, 佘跃惠, 黄金凤, 等.大庆油田聚合物驱后油藏微生物多样性研究[J].应用与环境生物学报, 2008, 14(5)：668-672.
[3] 包木太, 肖生科, 孔祥平, 等. 16S rRNA基因技术在油藏微生物生态研究中的应用[J].应用基础与工程科学学报, 2007, 15(3)：369-379.
[4] 王君, 马挺, 刘静, 等.利用PCR-DGGE技术指导高温油藏中功能微生物的分离[J].环境科学, 2008, 29(2)：462-468.
[5] Grabowski A, Nercessian O, Fayolle F, et al. Microbial diversity in production waters of a low temperature biodegraded oil reservoir[J]. FEMS Microbiology Ecology, 2005, 54(3)：427-443.
[6] Li H, Yang SZ, Mu BZ, et al. Molecular analysis of the bacterial community in a continental high-temperature and water-flooded petroleum reservoir[J]. FEMS Microbiology Letters, 2006, 257(1)：92-98.
[7] 佘跃惠, 张学礼, 张凡, 等.大港孔店油田水驱油藏微生物群落的分子分析[J].微生物学报, 2005, 45(3)：329-334.
[8] Orphan VJ, Taylor LT, Hafenbradl D, et al. Culture dependent and culture-independent characterization of icrobial assemblages associated with high-temperature petroleum reservoirs[J]. Applied Environmental Microbiology, 2000, 66(2)：700-711.
[9] 佘跃惠, 张凡, 向廷生, 等. PCR-DGGE 方法分析原油储层微生物群落结构及种群多样性[J]. 生态学报, 2005, 25(2)：238-242.
[10] 程海鹰, 肖生科, 马光东, 等.营养注入后油藏微生物群落16S rRNA基因的T-RFLP对比分析[J].石油勘探与开发, 2006, 33(3)：356-359.
[11] 程海鹰, 肖生科, 汪卫东, 等.变性梯度凝胶电泳方法在内源微生物驱油研究中的应用[J].石油学报, 2005, 26(6)：82-85.
[12] 赵凤敏. T-RFLP微生物监测技术及矿场应用研究[J].油气地质与采收率, 2005, 12(4)：64-66.
[13] 苏俊杰, 高光军, 宋永亭, 等.胜利油田单12区块内源微生物分子生态研究[J].石油钻采工艺, 2006, 28(1)：37-40.