Article(id=1241377723277430883, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1241377719049572379, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20230560, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1693497600000, receivedDateStr=2023-09-01, revisedDate=null, revisedDateStr=null, acceptedDate=1699545600000, acceptedDateStr=2023-11-10, onlineDate=1773897112912, onlineDateStr=2026-03-19, pubDate=1717430400000, pubDateStr=2024-06-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773897112912, onlineIssueDateStr=2026-03-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773897112912, creator=13701087609, updateTime=1773897112912, updator=13701087609, issue=Issue{id=1241377719049572379, tenantId=1146029695717560320, journalId=1192105938417971205, year='2024', volume='64', issue='6', pageStart='1691', pageEnd='2143', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1773897111904, creator=13701087609, updateTime=1773897665313, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1241380040286458828, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1241377719049572379, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1241380040286458829, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1241377719049572379, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1948, endPage=1959, ext={EN=ArticleExt(id=1241377723680084080, articleId=1241377723277430883, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Bacillus velezensis B6: mechanism of enhanced oil recovery and application in highly mineralized oil reservoirs, columnId=1241377722715394129, journalTitle=Acta Microbiologica Sinica, columnName=Geomicrobiological Applications, runingTitle=null, highlight=null, articleAbstract=

[Objective] To screen the indigenous functional microorganisms in reservoirs and explore their oil recovery mechanisms, we conducted field tests to determine the process and technical feasibility of enhancing crude oil recovery by indigenous microorganisms. [Methods] We collected samples from the Yingdong Oilfield and used oil plates to screen for the target bacteria. Next, we evaluated the environmental adaptability and optimized the culture conditions of the strain. Further, we measured the emulsifying, viscosity-reducing, paraffin-resistant, and hydrocarbon conversion abilities of the isolate to evaluate the oil recovery competence of the strain and explored its oil recovery mechanism. Finally, we carried out the microbial enhanced oil recovery tests in the oilfields. [Results] We isolatedBacillus velezensis B6 from the oil-water sample with an emulsifying activity index (EI24) of 100.00%, a viscosity reduction rate of 97.20%, and a paraffin resistance rate of 86.90%, which indicated that strain B6 had good emulsifying and viscosity-reducing abilities and improved oil recovery potential. Strain B6 could reduce heavy components in crude oil and increase light hydrocarbons to improve crude oil properties and quality. Moreover, we carried out single-well huff and puff and paraffin removal operations in Yingdong Oilfield, Yuejin Oilfield, and Huatugou Oilfield with a total of 62 well-times in the field test, which resulted in a cumulative oil increase of 1 460.36 tons and an average delay of 47 days in the well flushing cycle. The economic benefit was CNY 3.425 million, and the input-output ratio was 1:4. [Conclusion] The laboratory studies and field tests proved that indigenousB.velezensis B6 can significantly improve the oil recovery and well paraffin removal, with great application potential.

, correspAuthors=Peiyao SUN, authorNote=null, correspAuthorsNote=
*SUN Peiyao, Tel: +86-937-8920426, E-mail:
, copyrightStatement=Copyright ©2024 Acta Microbiologica Sinica. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Xianke CHEN, Peiyao SUN, Yong CAI, Miaofeng ZHANG, Wei ZHANG, Bo YANG, Anzhou MA), CN=ArticleExt(id=1241377726653845734, articleId=1241377723277430883, tenantId=1146029695717560320, journalId=1192105938417971205, language=CN, title=贝莱斯芽孢杆菌B6强化驱油机制分析及在高矿化度油藏中的应用, columnId=1241377722941886549, journalTitle=微生物学报, columnName=地质微生物应用, runingTitle=null, highlight=null, articleAbstract=

【目的】筛选油藏内源功能微生物并探究其驱油机理,现场试验以确定油藏内源微生物提高原油采收技术应用的工艺和技术可行性。【方法】采集英东油田样品,利用原油平板筛选驱油功能菌,评价其环境适应性并优化培养条件,通过乳化性能、降黏防蜡和烃转化能力等评估其驱油性能并探索其潜在机制,并在油田现场进行微生物强化驱油试验。【结果】从油水样品中分离到的贝莱斯芽孢杆菌(Bacillus velezensis) B6,其乳化活性指数(emulsifying activity index, EI24)值为100.00%,降黏率为97.20%,防蜡率为86.90%,表明菌株B6具有良好的乳化降黏性能,具有提高原油采收的潜力,并且能够减少原油中的重质组分而增加轻质烃类,改善原油物性,提升原油品质。进一步在英东油田、跃进油田和花土沟油田进行微生物单井吞吐和清防蜡作业,共进行62井次现场试验,累计增油1 460.36 t,平均延长洗井周期47 d,经济效益为342.50万元,投入产出比为1:4。【结论】通过室内研究和现场试验,证实内源微生物Bacillus velezensis B6具有显著提高原油采收和油井清防蜡的效果,应用潜力巨大。

, correspAuthors=孙培耀, authorNote=null, correspAuthorsNote=null, copyrightStatement=版权所有©《微生物学报》编辑部2024, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=PtjiWGnyXoCuCHR7h9M0EA==, magXml=6UkhP+zFyQ5LitCXm88yWQ==, pdfUrl=null, pdf=88M/TKBMfwiiiGAtAtzrtg==, pdfFileSize=948998, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=NEoeyMY9cv5+9MLlLxwGcg==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=+vk6Jd+Zn7Ko95qNoAKQWA==, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=陈显轲, 孙培耀, 蔡勇, 张淼峰, 张伟, 杨博, 马安周)}, authors=[Author(id=1241445029588291983, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1241377723277430883, orderNo=0, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1241445029705732506, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1241377723277430883, authorId=1241445029588291983, language=EN, stringName=Xianke CHEN, firstName=Xianke, middleName=null, lastName=CHEN, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=1, 2, 7, address=1 Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China
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A:pH. B:温度. C:矿化度. D:培养基对菌株B6生长的影响

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产液量(A)、产油量(B)和含水率(C)用于评估微生物单井吞吐效果,洗井周期(D)和油井载荷(E)用于评估清防蜡效果

, figureFileSmall=U9f9keCqSVZkhKIIxikzKA==, figureFileBig=IFlND91IyBfn2RXdMuznLQ==, tableContent=null), ArticleFig(id=1241445038920618794, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1241377723277430883, language=EN, label=Table 1, caption=

Evaluation of oil recovery performance of strain B6 in different media

, figureFileSmall=null, figureFileBig=null, tableContent=
MediaSurface tension (mN/m)EI24 (%)Paraffin control rate (%)Viscosity reduction rate (%)
Improved LB medium27.9596.5486.9095.08
Fermentation medium27.33100.0074.0097.20
), ArticleFig(id=1241445039054836528, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1241377723277430883, language=CN, label=表1, caption=

不同培养基中菌株B6的驱油性能评价

, figureFileSmall=null, figureFileBig=null, tableContent=
MediaSurface tension (mN/m)EI24 (%)Paraffin control rate (%)Viscosity reduction rate (%)
Improved LB medium27.9596.5486.9095.08
Fermentation medium27.33100.0074.0097.20
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贝莱斯芽孢杆菌B6强化驱油机制分析及在高矿化度油藏中的应用
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陈显轲 1, 2, 7 , 孙培耀 3, * , 蔡勇 4 , 张淼峰 5 , 张伟 5 , 杨博 6 , 马安周 1, 7
微生物学报 | 地质微生物应用 2024,64(6): 1948-1959
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微生物学报 | 地质微生物应用 2024, 64(6): 1948-1959
贝莱斯芽孢杆菌B6强化驱油机制分析及在高矿化度油藏中的应用
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陈显轲1, 2, 7, 孙培耀3, * , 蔡勇4, 张淼峰5, 张伟5, 杨博6, 马安周1, 7
作者信息
  • 1 中国科学院生态环境研究中心, 北京 100085
  • 2 中国科学院大学中丹学院, 北京 101400
  • 3 青海油田分公司钻采工艺研究院, 甘肃 敦煌 736202
  • 4 青海油田分公司采油五厂, 青海 茫崖 816400
  • 5 青海油田分公司采油三厂, 青海 茫崖 816400
  • 6 青海油田分公司采油一厂, 青海 茫崖 816400
  • 7 中国科学院大学, 北京 100049
Bacillus velezensis B6: mechanism of enhanced oil recovery and application in highly mineralized oil reservoirs
Xianke CHEN1, 2, 7, Peiyao SUN3, * , Yong CAI4, Miaofeng ZHANG5, Wei ZHANG5, Bo YANG6, Anzhou MA1, 7
Affiliations
  • 1 Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China
  • 2 Sino-Danish College, University of Chinese Academy of Sciences, Beijing 101400, China
  • 3 Drilling and Production Technology Research Institute of Qinghai Oilfield Company, Dunhuang 736202, Gansu, China
  • 4 The Fifth Oil Production Plant of Qinghai Oilfield Company, Mangya 816400, Qinghai, China
  • 5 The Third Oil Production Plant of Qinghai Oilfield Company, Mangya 816400, Qinghai, China
  • 6 The First Oil Production Plant of Qinghai Oilfield Company, Mangya 816400, Qinghai, China
  • 7 University of Chinese Academy of Sciences, Beijing 100049, China
出版时间: 2024-06-04 doi: 10.13343/j.cnki.wsxb.20230560
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【目的】筛选油藏内源功能微生物并探究其驱油机理,现场试验以确定油藏内源微生物提高原油采收技术应用的工艺和技术可行性。【方法】采集英东油田样品,利用原油平板筛选驱油功能菌,评价其环境适应性并优化培养条件,通过乳化性能、降黏防蜡和烃转化能力等评估其驱油性能并探索其潜在机制,并在油田现场进行微生物强化驱油试验。【结果】从油水样品中分离到的贝莱斯芽孢杆菌(Bacillus velezensis) B6,其乳化活性指数(emulsifying activity index, EI24)值为100.00%,降黏率为97.20%,防蜡率为86.90%,表明菌株B6具有良好的乳化降黏性能,具有提高原油采收的潜力,并且能够减少原油中的重质组分而增加轻质烃类,改善原油物性,提升原油品质。进一步在英东油田、跃进油田和花土沟油田进行微生物单井吞吐和清防蜡作业,共进行62井次现场试验,累计增油1 460.36 t,平均延长洗井周期47 d,经济效益为342.50万元,投入产出比为1:4。【结论】通过室内研究和现场试验,证实内源微生物Bacillus velezensis B6具有显著提高原油采收和油井清防蜡的效果,应用潜力巨大。

微生物吞吐采油  /  乳化降黏  /  规模化发酵  /  微生物清防蜡

[Objective] To screen the indigenous functional microorganisms in reservoirs and explore their oil recovery mechanisms, we conducted field tests to determine the process and technical feasibility of enhancing crude oil recovery by indigenous microorganisms. [Methods] We collected samples from the Yingdong Oilfield and used oil plates to screen for the target bacteria. Next, we evaluated the environmental adaptability and optimized the culture conditions of the strain. Further, we measured the emulsifying, viscosity-reducing, paraffin-resistant, and hydrocarbon conversion abilities of the isolate to evaluate the oil recovery competence of the strain and explored its oil recovery mechanism. Finally, we carried out the microbial enhanced oil recovery tests in the oilfields. [Results] We isolatedBacillus velezensis B6 from the oil-water sample with an emulsifying activity index (EI24) of 100.00%, a viscosity reduction rate of 97.20%, and a paraffin resistance rate of 86.90%, which indicated that strain B6 had good emulsifying and viscosity-reducing abilities and improved oil recovery potential. Strain B6 could reduce heavy components in crude oil and increase light hydrocarbons to improve crude oil properties and quality. Moreover, we carried out single-well huff and puff and paraffin removal operations in Yingdong Oilfield, Yuejin Oilfield, and Huatugou Oilfield with a total of 62 well-times in the field test, which resulted in a cumulative oil increase of 1 460.36 tons and an average delay of 47 days in the well flushing cycle. The economic benefit was CNY 3.425 million, and the input-output ratio was 1:4. [Conclusion] The laboratory studies and field tests proved that indigenousB.velezensis B6 can significantly improve the oil recovery and well paraffin removal, with great application potential.

microbial huff and puff  /  emulsification and viscosity reduction  /  large-scale fermentation  /  microbial paraffin removal
陈显轲, 孙培耀, 蔡勇, 张淼峰, 张伟, 杨博, 马安周. 贝莱斯芽孢杆菌B6强化驱油机制分析及在高矿化度油藏中的应用. 微生物学报, 2024 , 64 (6) : 1948 -1959 . DOI: 10.13343/j.cnki.wsxb.20230560
Xianke CHEN, Peiyao SUN, Yong CAI, Miaofeng ZHANG, Wei ZHANG, Bo YANG, Anzhou MA. Bacillus velezensis B6: mechanism of enhanced oil recovery and application in highly mineralized oil reservoirs[J]. Acta Microbiologica Sinica, 2024 , 64 (6) : 1948 -1959 . DOI: 10.13343/j.cnki.wsxb.20230560
石油作为当今社会的重要能源之一,在世界经济和人类社会的发展中起着至关重要的作用。自工业革命以来,随着石油化工行业的深入发展,全球石油需求量不断增加,且原油价格持续上涨[1-2]。但是,随着油田开采年限的增加,常规易采石油资源储量越来越少,原油采收率越来越低。目前地下油藏环境中仍有大量原油亟待有效开采,其残留的剩余原油占60%–70%[3-5]。因此,如何绿色、高效、经济地开采出地下剩余原油不仅是能源领域的研究热点,而且事关国家经济发展和社会稳定。
微生物强化采油(microbial enhanced oil recovery, MEOR)是随着3次采油而发展的新兴技术,具有工艺简单、成本低廉和环境友好等特点,在国内外被广泛推广使用[6-11],是最具有发展前景的采油技术之一。微生物强化采油技术工艺主要包括微生物单井吞吐、微生物清防蜡、微生物驱替和微生物选择性封堵,同时也开发了基于微生物强化采油的多种工艺的整合,如多轮吞吐、清防蜡耦合单井吞吐等驱油技术。近20余年来,筛选了大量高产生物表面活性剂或石油烃降解等的微生物,绝大多数菌株已在室内研究中证实了其具有驱油的性能,为提高剩余原油的采收奠定了坚实的基础,但目前现场试验验证和应用仍严重不足[12]。目前,胜利油田和大庆油田等是我国微生物强化驱油技术现场应用较为成熟且取得成果的油田[13-16]。然而,油藏地层环境具有明显的地质特异性、非均质性强,油藏模式复杂多样,这导致微生物强化采油技术的具体应用表现出一定的特异性,因而需要依据油藏环境和原油物性施用合适的驱油微生物。
针对青海油田地层油藏环境矿化度较高和原油采出率较低等现实问题,本文通过对筛选获得的内源微生物贝莱斯芽孢杆菌(Bacillus velezensis) B6生长及其功能特征的研究,从而评估该菌株对原油的作用效果。该菌株经规模化培养后在英东油田、跃进油田和花土沟油田开展微生物单井吞吐和清防蜡试验,实现原位乳化降黏和井筒清防蜡,提高原油流动性并增加油井洗井周期,验证其实际应用价值。本项研究为开发适用于较高矿化度油藏中稠油冷采的绿色生物采油体系提供了科学依据和技术支撑。
葡萄糖、石蜡油、琼脂粉、氯化钠、磷酸氢二钾和七水合硫酸亚铁等分析纯试剂购自国药集团化学试剂有限公司;正己烷等色谱纯有机试剂购自Thermo Fisher Scientific公司。气相色谱-质谱联用仪(岛津公司),多功能酶标仪(SpectraMax i3x,Molecular Devices公司),表面张力仪(Lauda Scientific公司),流变仪(HAAKETM ViscotesterTM iQ,Thermo Fisher Scientific公司),光学显微镜(Olympus公司)。现场作业使用泵车(2.5×104 Pa)、罐车(30 m3)等专业设备。
使用10 L无菌塑料桶,在英东油田油井工作过程中采集油水样,在4 ℃条件下送至实验室进行下一步分析。
原油培养基(g/L):原油20,NH4Cl 0.5,NaNO3 2,K2HPO4 1,FeSO4·7H2O 0.05,SDS 0.01,pH 6.5–7.5。固体原油培养基中加入琼脂粉8 g/L。115 ℃灭菌45 min。
改良LB培养基(Medium A) (g/L):胰蛋白胨10,酵母粉5,NaCl 20,FeSO4·7H2O 0.05,pH 6.8–7.2。115 ℃灭菌30 min。
发酵培养基(Medium B) (g/L):葡萄糖30,酵母粉4,胰蛋白胨8,K2HPO4 2,NaCl 20,pH 6.8–7.2。115 ℃灭菌30 min。
培养条件:37 ℃,150–200 r/min。
将油水样接种到原油培养基中培养10 d后采用稀释平板法进行驱油功能微生物的分离纯化。取100 μL稀释度为10−7、10−8和10−9的菌液涂布于固体原油培养基上,挑选平板上具有不同菌落形态和较大透明圈的菌株作为实验菌株,再次进行稀释涂布培养。随后,挑取单菌落转接到改良LB培养基中培养24 h,使用通用引物27F/1492R扩增16S rRNA基因全长序列[17],由金唯智生物科技(北京)有限公司测序后基于基因信息确定菌株的分类地位。
配制发酵培养基,使用HCl或NaOH调节培养基的pH分别为6.0、7.0、8.0和9.0,OD600为1的菌液接种量为1%,37 ℃、200 r/min振荡培养,定期取样测量菌液在600 nm处的光密度值。
配制发酵培养基,在30、37、40 ℃和50 ℃条件下培养,OD600为1的菌液接种量为1%,37 ℃、200 r/min振荡培养,定期取样测量菌液在600 nm处的光密度值。
配制NaCl浓度分别为20、100和150 g/L的发酵培养基,OD600为1的菌液接种量为1%,37 ℃、200 r/min振荡培养,定期取样测量菌液在600 nm处的光密度值。
通过乳化活性指数(emulsifying activity index, EI24)对菌液进行乳化性能评价[18]。以石蜡油作为原油替代物,分别各取2 mL石蜡油与菌株上清发酵液加入到10 mL试管中,并记录上层有机相总高度。使用漩涡振荡器振荡约3 min,混匀后室温静置24 h,记录白色乳化层高度,并计算EI24值。以培养基作空白对照,设置3组平行。其中,EI24=乳化层高度/有机层总高度×100%。
使用流变仪测定菌株B6处理前后37 ℃条件下的原油黏度,采用倒扣瓶法评估菌株B6对原油的防蜡率,原油黏度和防蜡率的具体操作方法参考文献[18]。其中,原油黏度为2 630.42 mPas,含蜡量为25.00%。
采用悬滴法测定菌株B6发酵上清液的表面张力[19]。取40 mL发酵菌液,10 000×g室温离心10 min,取发酵上清液,利用表面张力仪测定菌株发酵液的表面张力,测定3次,取平均值。
处理后的原油样品使用超声萃取法提取,正己烷稀释后使用气相色谱-质谱联用仪(gaschromatography-mass spectrometry, GC-MS)进行分析,详细的处理和操作方法参考文献[20]。
采用薄层色谱-火焰离子化检测(thin-layer chromatography with flame ionization detection, TLC-FID)技术对残余油样品中的烷烃、芳香烃、树脂和沥青质4种组分含量进行分析[21]
现场微生物单井吞吐和清防蜡试验油井井深1 121–1 767 m,油藏温度34–53 ℃,平均油井热洗周期45 d。单井储层孔隙度15%–24%,渗透率30–89 md。原油黏度2 000–4 000 mPas,地层水矿化度为100 000–140 000 mg/L,平均pH为8.5。
规模化发酵:配制发酵培养基,规模化发酵参数如下:转速80–150 r/min,37℃,压力0.3–0.5 MPa,通气比0.3–0.5 VVM。
单井吞吐作业:单井吞吐施工液中菌液浓度为15%,营养液浓度为6%,pH约为8.0。根据试验油井油层厚度、孔隙度和处理半径最终确定注入施工液量为6–10 m3。使用套管环空注入法,从油套环空间注入单井吞吐施工液,注入排量控制在0.2–0.3 m3/min,注入压力控制在1.5×104 Pa内,在注入过程中根据注入泵压等动态参数的变化情况调整注入速度和注入量。待全部注入后,关井7 d后开井作业,定期监测油井产量。
清防蜡作业:清防蜡作业液中菌液浓度为30%,营养液浓度为8%,pH约为8.5。采用套筒注入方式,注入量为1 m3,注入完成后油井正常生产,定期监测油井示功图,评估洗井周期。
使用GraphPad Prism 8软件进行统计分析和图示化,误差棒表示标准差。
基于在线工具NCBI BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi)和EZBioCloud (www.ezbiocloud.net/identify)对筛选获得的驱油功能菌株B6的16S rRNA基因全长序列进行比对分析,结果表明菌株B6属于Bacillus属,Bacillus velezensis种。使用MEGA 11.0[22]构建进化树后使用iTOL (https://itol.embl.de/)对进化树进行美化调整。菌株B6与B.velezensis CR-502、B.velezensis JK19和B.velezensis JS25R等亲缘关系更近(图1),命名为B.velezensis B6,NCBI序列号为OR277460。国内外关于B.velezensis的研究主要集中在拮抗动植物病原菌,促进动植物生长等方面[23-26]。最近的一篇报道表明B.velezensis由于其高产生物表面活性剂而在微生物强化驱油中取得了良好的应用效果,但是其环境适应性和驱油功能特性与菌株B6存在不同[27]。此外,与菌株B6亲缘关系更近的B.velezensis SRCM102755等菌株基因组中均含有生物表面活性剂合成基因srfAAsrfABsrfAC[28]。因此,该菌在微生物强化采油中可能具有重要作用。
根据生长曲线(图2),B.velezensis B6在培养过程中表现出较短的生长周期,培养5 h即可达到对数期,12 h后逐渐到达生长稳定期。在pH 6.0–8.0时菌株B6表现出良好的生长趋势,而在pH 9.0时,B6则几乎不生长(图2A),且与B.velezensis BSA1相比在pH 8.0时能更快地到达生长稳定期[27];菌株B6的生长能力随着培养温度的升高表现出先增强后降低的趋势,30 ℃延缓了其对数期(图2B);在100 g/L以上的矿化环境中,菌株B6的生长受到了一定的限制(图2C);菌株B6在发酵培养基中比在改良LB培养基中表现出较高的生长优势(图2D)。综上所述,B.velezensis B6在pH 6.0–8.0、30–40 ℃的环境中能够良好生长,在100 g/L的矿化度以上生长受到一定抑制,说明本研究分离得到的菌株B6能够较好地适应油田油藏环境,具有实际应用的潜在价值。
菌液具有较低的表面张力和较高的乳化能力能在提高原油采收率方面起着重要作用[29]。如表1所示,菌株B6发酵上清液的表面张力(27.33 mN/m)显著低于纯水的表面张力(72.80 mN/m),良好的表面活性剂能够显著降低表面张力,可以有效改变油、岩、水的界面状态,有助于提高地层中油相的流动性,从而提高驱油效率[30]。将发酵上清液与液体石蜡混合后,EI24值为100.00%,具有优良的乳化性能,并高于先前单菌或混合菌系的研究结果。如,Liu等[31]测定的地衣芽孢杆菌(Bacillus licheniformis) L20发酵上清液的EI24值为62.00%,魏晓霞等[18]测定了来源于青海油田的混合菌系QZ-10的EI24值为91.11%。因此,以上结果表明菌株B6产生的生物表面活性剂其性能更为突出,具有较高的应用于微生物强化驱油的潜力。
此外,经菌株B6处理后,倒扣瓶法测得的其防蜡率均高于70.00%,在37 ℃条件下降黏率最高可达到97.20% (表1)。王卫强等[32]从石油污染土壤中分离筛选获得的一株假单胞菌(Pseudomonas sp.) W12#,其防蜡率为34.66%,降黏率为63.75%,表明菌株B6在防蜡和降黏等方面一定程度上优于先前的研究。此外,菌株B6使用发酵培养基进行发酵培养时,其乳化和降黏性能优于改良LB培养基(表1),表明大规模培养菌株B6时使用发酵培养基可能具有较好的驱油效果。综上所述,B.velezensis B6的驱油性能不仅强于单菌而且优于混合菌系,具有强化驱油的潜力,环境适应性良好,可用于微生物单井吞吐和油井清防蜡作业。
原油通常是一种化学成分复杂的黑色易燃液体,根据其在有机溶剂中的溶解度不同,可分为饱和烃、芳香烃、树脂和沥青质[33]。使用发酵培养基对菌株B6处理前后的原油进行四组分测试,各组分含量如图3所示。经菌株B6处理后,原油中的饱和烃含量显著增加了4.38% (P<0.000 1),芳香烃、树脂和沥青质含量减少,其中沥青质在处理前后含量差异不大(图3)。这可能是该菌较强的乳化性能提高了细菌对疏水性烃类的可利用性,从而提高了长链或大分子多环烃的降解[34-35]。此外,沥青质等大分子是原油高黏性和油井堵塞的主要原因[36],该菌在一定程度上能够降解该类大分子物质。因此,表明菌株B6能够将原油中的大分子重质物质,如树脂和沥青质,降解成小分子轻质石油基组分,增加了原油流动性,提高了原油的品质。
对菌株B6处理前后原油中的饱和烃(C10−C40)进行检测,分析处理前后不同饱和烃组分的含量变化,以探讨对原油物性的影响。与空白组相比,使用改良LB培养基培养时,C10−C20的含量增加了5.60%,C21−C30和C31−C40的含量分别减少了4.18%和1.42%;使用发酵培养基培养时,C10−C20的含量增加了6.28%,C21−C30和C31−C40的含量分别减少了4.60%和1.68%。随着碳数的增加,菌株B6处理后原油中的饱和烃降解率呈现下降趋势,特别是C30−C40范围内的烃类物质(图4)。结果表明,菌株B6能够对长链饱和烃进行分解,因此原油中大分子组分的减少和短链烃的增加有效地降低了原油的黏度,改变了原油物性。此外,当碳数高于20时,易出现结蜡现象,经处理后碳数高于20的长链饱和烃明显减少,这有助于减缓油井结蜡现象,延长洗井周期[37]。综上所述,B.velezensis B6能够降解原油中的大分子重质组分和长链烃等物质,表明该菌在油田微生物驱油和油井清防蜡中具有良好的应用前景。
微生物单井吞吐具有见效快、操作简单等优点,是常用的一种微生物驱油工艺,且一般会使用多轮吞吐操作以提高其有效期和原油采出率。微生物单井吞吐和清防蜡作业选用的是以产生生物表面活性剂为主的B.velezensis B6及其发酵产物的混合物,其混合液pH值为6.6–7.5,每毫升菌数≥108个。在现场作业前,对菌液进行乳化性能测试,EI24值均为100.00%。结合室内研究结果和油藏环境以及先前操作经验[27],在英东油田、跃进油田和花土沟油田进行微生物单井吞吐和清防蜡试验。其中,微生物单井吞吐主要评价指标为油井的日产液量、日产油量和含水率,而微生物油井清防蜡则以洗井周期和油井载荷为主要评价指标(图5)。
图5A5C所示,微生物单井吞吐作业前油井的平均日产液量为4.12 t,日产油量为1.05 t,含水率65.61%;作业后油井的平均日产液量为5.02 t,日产油量为1.56 t,含水率60.05%,微生物单井吞吐有效率100%。与作业前相比,平均日产液增加0.9 t,平均日增油0.51 t,平均含水率降低了5.56%,作业后驱油效果显著,且采油率高于枯草芽孢杆菌(Bacillus subtilis) XT-1和螯台球菌(Chelatococcus daeguensis) HB-4岩芯驱油的结果[38-39],表明使用油藏内源微生物B.velezensis B6的强化驱油效果更为显著。先前的研究表明,注入菌液制剂可以激活油藏中的其他功能微生物,如产表面活性剂、石油烃降解和产气体的微生物,不同功能微生物的综合作用有助于提高石油采收率[27]。此外,吞吐油井能够有效延长洗井周期,这可能归功于微生物对大分子重质原油组分的降解转化,原油物性得到了改善。在油井清防蜡现场试验中,处理前平均洗井周期为45 d,处理后平均洗井周期为92 d,平均延长洗井周期47 d (图5D),清防蜡有效率96.55%,优于QZ-10混合菌液的作业效果[18]。作业前油井最大载荷与最小载荷差值的平均值为19.68 kN,作业后油井最大载荷与最小载荷差值的平均值为17.38 kN,作业后油井载荷显著降低(图5E)。上述相关数据表明该菌能够有效地改善油井载荷,降低采油负荷。综上所述,B.velezensis B6具有良好的现场应用效果,在提高原油采收率方面具有很大的实际应用价值。
在英东油田、跃进油田和花土沟油田进行微生物单井吞吐和清防蜡作业,共进行62井次现场试验。微生物单井吞吐作业后平均日增液量最大值4.32 t,平均日增油最大值1.97 t,最大有效期138 d;作业后平均日增液量最小值1.29 t,平均日增油最小值0.32 t,最小有效期35 d。吞吐前后对比,产液量累计增加了1 613.22 t,产油量累计增加了1 124.85 t,平均有效期128 d。原油结算价格按3 000元/t计算,经济效益为249.00万元,投入产出比为1:3.8。清防蜡作业后累计增油335.51 t,平均延长洗井周期47 d,经济效益为93.50万元,投入产出比为1:4.8。
在地下油藏环境中,微生物生长代谢过程中所产生的生物活性物质具有改善油藏环境、乳化原油、降低原油黏度和改变原油物性等功能,从而提高残余原油的采收率[40]。本研究从英东油田油水样中筛选到一株能够产生生物表面活性剂的B.velezensis B6,该菌具有适应较高矿化度油藏环境的能力,表现出优良的乳化降黏性能。此外,通过对处理前后原油组分的分析,表明菌株B6能够降低大分子重质石油基组分,如树脂和沥青质,增加了小分子短链饱和烃等轻质烃组分,改善原油物性,提高了原油的流动性和原油的品质。规模化发酵的菌液在英东油田、跃进油田和花土沟油田进行微生物单井吞吐和清防蜡试验,现场试验效果良好,累计增油1 456.36 t,平均延长洗井周期47 d,经济效益为342.50万元,投入产出比为1:4。综上所述,B.velezensis B6能够较好地应用于微生物单井吞吐和清防蜡等微生物强化采油。但是,针对清防蜡作业仍需要优化工艺参数,包括培养基的选择、菌液的注入量和注入方式,以提高现场作业效率。此外,鉴于油井层间较强的非均质性,下一步可通过组学技术、原油指纹图谱和驱油模型等多角度的交叉分析,了解并确定改善原油物性的关键机制和因素,并完善长期原油采收监测技术,建立适合多层间的油井组微生物强化驱油工艺体系。
  • 国家重点研发计划(2018YFA0901200)
  • 青海油田微生物采油技术推广应用(2022T01)
  • 青海省科技计划(2022-QY-202)
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2024年第64卷第6期
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doi: 10.13343/j.cnki.wsxb.20230560
  • 接收时间:2023-09-01
  • 首发时间:2026-03-19
  • 出版时间:2024-06-04
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  • 收稿日期:2023-09-01
  • 录用日期:2023-11-10
基金
National Key Research and Development Program of China(2018YFA0901200)
国家重点研发计划(2018YFA0901200)
Extension and Application of Microbial Oil Recovery Technology in Qinghai Oilfield(2022T01)
青海油田微生物采油技术推广应用(2022T01)
Qinghai Science and Technology Plan(2022-QY-202)
青海省科技计划(2022-QY-202)
作者信息
    1 中国科学院生态环境研究中心, 北京 100085
    2 中国科学院大学中丹学院, 北京 101400
    3 青海油田分公司钻采工艺研究院, 甘肃 敦煌 736202
    4 青海油田分公司采油五厂, 青海 茫崖 816400
    5 青海油田分公司采油三厂, 青海 茫崖 816400
    6 青海油田分公司采油一厂, 青海 茫崖 816400
    7 中国科学院大学, 北京 100049

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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