三次采油菌配伍发酵性能的研究
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摘要
微生物提高石油采收率技术(Microbial Enhanced Oil Recovery Technology, MEOR)是一项适用范围广、经济效益好、工艺简单、无污染的综合性三次采油技术。其主要机理是利用微生物及其代谢产物作用于原油,从而降低原油黏度与油水间界面张力以改善原油在油藏中的流动性,最终提高驱油效率。
本实验对黑龙江大学生命科学学院采油微生物实验室保存的菌种和从大庆油田油污水样与土样中分离的菌种进行初筛,得到10株单一代谢物产量高的细菌。对筛选出的10株菌做原油乳化和降黏实验,选出6株采油性能好的菌株并在不同温度、酸碱度、盐度条件下检测细菌生长情况。
选取无拮抗作用的菌株进行配伍发酵实验,得到环境适应能力及原油降黏能力均好的一组配伍混合菌2-2、S2-1、YG1-2,其原油降黏率为76.80%,在温度35-50℃、酸碱度pH5-9、盐度0.3%-3%的范围内菌浓度OD_(600) >2.0,混合菌有机酸产量为2.907 g/L,脂肽产量为803.6mg/L。
结合细菌的形态学观察、生理生化特征实验及16SrDNA序列分析,鉴定三株菌分别为:2-2枯草芽孢杆菌(Bacillus subtilis)、S2-1苏云金芽孢杆菌(Bacillus thuringiensis)、YG1-2蜡样芽孢杆菌(Bacillus cereus)。发酵条件经正交实验优化后,配伍菌的原油降黏率达到83.46%,较优化前提高了8.67%。
实验结果表明,配伍混合菌在耐温性,耐盐性,耐酸碱性及原油的降黏率上均优于单菌,具有配伍优势。
Microbial Enhanced Oil Recovery was a comprehensive EOR technique, which wide application, good economical returns, simple process, no pollution. The main mechanism is the use of microbes and their metabolites acting on the crude oil, thereby reduce oil viscosity and interfacial tension between oil and water in order to improve the flow of crude oil in the reservoir, finally improve the efficiency of oil displacement.
In this experiment, first, strains which are presered of the laboratory and collectded Daqing Oilfield waste water samples and soil samples are screened. 10 strains of high yield of Single metabolite are obtained, do viscosity reduction of crude oil emulsion test for 10 strains. Selected 6 strains which have good effect in MEOR detection growth of strains under different temperatures, acidities, salinities.
Selected no antagonistic strains to do fermentation compatibility test, obtain environmental adaptability and the viscosity reduction of crude oil capacity both good compatible of strains 2-2、S2-1、YG1-2,crude oil viscosity reduction rate of 76.80%,Temperature 35-50℃, pH 5-9, salinity range of 0.3% -3% of the strains to maintain the concentration of OD_(600) > 2.0 mixed strains production of organic acid is 2.907 g / L,Lipopeptide yield 803.6mg / L.
After the strain morphological, physiological and biochemical characteristics test and identification of 16SrDNA,Three strains were: 2-2 (Bacillus subtilis), S2-1 (Bacillus thuringiensis), YG1-2 (Bacillus cereus). Fermentation conditions optimized by orthogonal test, Compatibility of strains of crude oil viscosity reduction rate 83.46%, than before optimization increased 8.67%.
The results show that compatibility of mixed strains on the temperature resistance, salt resistance, acid resistance and viscosity reduction of crude oil are better than single strain, have advantages of compatibility.
本实验对黑龙江大学生命科学学院采油微生物实验室保存的菌种和从大庆油田油污水样与土样中分离的菌种进行初筛,得到10株单一代谢物产量高的细菌。对筛选出的10株菌做原油乳化和降黏实验,选出6株采油性能好的菌株并在不同温度、酸碱度、盐度条件下检测细菌生长情况。
选取无拮抗作用的菌株进行配伍发酵实验,得到环境适应能力及原油降黏能力均好的一组配伍混合菌2-2、S2-1、YG1-2,其原油降黏率为76.80%,在温度35-50℃、酸碱度pH5-9、盐度0.3%-3%的范围内菌浓度OD_(600) >2.0,混合菌有机酸产量为2.907 g/L,脂肽产量为803.6mg/L。
结合细菌的形态学观察、生理生化特征实验及16SrDNA序列分析,鉴定三株菌分别为:2-2枯草芽孢杆菌(Bacillus subtilis)、S2-1苏云金芽孢杆菌(Bacillus thuringiensis)、YG1-2蜡样芽孢杆菌(Bacillus cereus)。发酵条件经正交实验优化后,配伍菌的原油降黏率达到83.46%,较优化前提高了8.67%。
实验结果表明,配伍混合菌在耐温性,耐盐性,耐酸碱性及原油的降黏率上均优于单菌,具有配伍优势。
Microbial Enhanced Oil Recovery was a comprehensive EOR technique, which wide application, good economical returns, simple process, no pollution. The main mechanism is the use of microbes and their metabolites acting on the crude oil, thereby reduce oil viscosity and interfacial tension between oil and water in order to improve the flow of crude oil in the reservoir, finally improve the efficiency of oil displacement.
In this experiment, first, strains which are presered of the laboratory and collectded Daqing Oilfield waste water samples and soil samples are screened. 10 strains of high yield of Single metabolite are obtained, do viscosity reduction of crude oil emulsion test for 10 strains. Selected 6 strains which have good effect in MEOR detection growth of strains under different temperatures, acidities, salinities.
Selected no antagonistic strains to do fermentation compatibility test, obtain environmental adaptability and the viscosity reduction of crude oil capacity both good compatible of strains 2-2、S2-1、YG1-2,crude oil viscosity reduction rate of 76.80%,Temperature 35-50℃, pH 5-9, salinity range of 0.3% -3% of the strains to maintain the concentration of OD_(600) > 2.0 mixed strains production of organic acid is 2.907 g / L,Lipopeptide yield 803.6mg / L.
After the strain morphological, physiological and biochemical characteristics test and identification of 16SrDNA,Three strains were: 2-2 (Bacillus subtilis), S2-1 (Bacillus thuringiensis), YG1-2 (Bacillus cereus). Fermentation conditions optimized by orthogonal test, Compatibility of strains of crude oil viscosity reduction rate 83.46%, than before optimization increased 8.67%.
The results show that compatibility of mixed strains on the temperature resistance, salt resistance, acid resistance and viscosity reduction of crude oil are better than single strain, have advantages of compatibility.
引文
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[4] Clark J B et al.In situ microbial enhancement of oil production [J].Industry Microbiology,1981, 15:695-701.
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[6]吴坛珍,郑占,曹煜等.微生物提高采出率的室内研究[J].生物加工过程,2003,1(2):53-58.
[7]程海鹰.油藏微生物在多孔介质中的运移过程研究[D].博士论文,2006
[8]周大林,马东,梁文利等.微生物采油技术研究[J].内蒙古石油化工,2008,(10):203-206.
[9]王惠,卢渊,伊向艺.国内外微生物采油技术综述[J].大庆石油地质与开发, 2003, 22(5):49
[10]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.
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[16] Steven Bryant.Microbial Enhanced Oil Recovery: A Sober Look at an Infectious Idea.SPE101452-DL,2001.
[17]王修垣.石油微生物学在中国科学院微生物研究所的发展[J].微生物学通报DEC 20, 2008, 35(12): 1851-1861
[18]王惠,卢渊.微生物技术在石油工业中的应用[J].科技进步与对策,2003,增刊:291-292
[19]王惠,卢渊,伊向艺.国内外微生物采油技术综述[J].大庆石油地质与开发2003, 22(5)49
[20] Smith T L, Gabriela Trebbau.微生物强化采油使重质原油产量锐增[J].国外油田工程, 1993, (3):12-13
[21]罗新华,方辉.在沙捞越Bokor油田实施的微生物提高采收率技术[J].国外油田工程, 2002, 18(10): l-6
[22] Matsumae H, Furui M, Shibatani T, etal.Production of oPtically aetive Phenylglycidic aeid ester by the lipase from serratia marceseens on a hollow-fiber Membrane reactor[J]. Ferment&Bioeng, 1994, 78(l): 59-63
[23]邹少兰,刘如林.内源微生物采油技术的历史与现状[J].微生物学通报,2002,29(5): 70-73
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