石油降解菌的筛选、鉴定及菌群构建

低温石油降解菌LHB16的筛选及降解特性研究•相关推荐
低温石油降解菌LHB16的筛选及降解特性研究
摘要：目的:筛选、鉴定低温石油降解菌并对其降解特性进行研究.方法:富集分离低温石油降解菌;采用形态学、生理生化实验和分子生物学方法进行菌种鉴定;紫外分光光度法和GC-MS检测石油降解特性.结果:自盘锦油田低温环境土样中分离到1株低温菌,命名为LHB16,该菌能以石油烃为惟一碳源和能源.经鉴定为嗜麦芽窄食单胞菌(Stenotrophomonas maltophilia).该菌生长温度范围0℃～35℃,最适生长温度15℃.在接种量为2%(V/V),原油浓度为0.5%(W/V),振荡培养10 d时,降解率可达80.16%.石油中长链烷烃C15～C32被完全降解.传代培养数代,降解率为81.06%,降解性能稳定.结论:菌株LHB16在低温地区石油污染的`生物治理中有良好的应用前景.作者：李兵张庆芳窦少华孙子羽王宇迟乃玉 LI Bing ZHANG Qing-fang DOU Shao-hua SUN Zi-yu WANG Yu CHI Nai-yu 作者单位：李兵,张庆芳,窦少华,孙子羽,迟乃玉,LI Bing,ZHANG Qing-fang,DOU Shao-hua,SUN Zi-yu,CHI Nai-yu(大连大学生物工程学院,辽宁大连,116622)
王宇,WANG Yu(大连轻工业大学生物与食品工程学院,辽宁大连,116034)
期刊：生物技术 PKU Journal：BIOTECHNOLOGY 年，卷(期)：2010, 20(5) 分类号：X172 关键词：低温石油降解筛选 Stenotrophomonas maltophilia。

山东轻工业学院2012届本科生毕业论文目录摘要············································································································ - 3 -ABSTRACT ······································································································ - 4 -第一章绪论··································································································· - 5 -1.1．微生物提高采收率的研究状况······················································ - 5 -1.1.1．方法的提出··········································································· - 5 -1.1.2．国内研究现状[1] ··································································· - 6 -1.1.3．国外研究现状······································································· - 6 -1.2．微生物采油的机理及条件······························································ - 7 -1.2.1．微生物采油方法的机理······················································· - 7 -1.2.2.微生物采油的条件·································································· - 8 -1.2.2.1.菌种的选择··········································································· - 8 -1.2.2.2.油藏的条件··········································································· - 8 -1.3.常用的MEOR方法··········································································· - 9 -1.3.1．应用微生物表面活性剂助采··············································· - 9 -1.3.1.1.生物表面活性剂··································································· - 9 -1.3.1.2.生物表面活性剂在石油工业中的应用······························· - 9 -1.3.2．生产井周期性微生物处理··················································· - 9 -1.3.3．微生物注水········································································· - 10 -1.3.4．活化油层内源有微生物群落············································· - 10 -1.4．微生物采油的优势与不足···························································· - 10 -1.5．本论文的研究内容········································································ - 10 -第二章原油降解假单胞菌的分离筛选与鉴定········································· - 11 -2．1．菌种筛选····················································································· - 11 -2．1．1．材料··············································································· - 11 -2．1．2．方法··············································································· - 11 -2．1．3．结果与讨论··································································· - 12 -2．2．菌种鉴定····················································································· - 12 -2．2．1．材料··············································································· - 12 -2．2．1．1．菌株········································································· - 12 -2．2．1．2．培养基····································································· - 12 -2．2．1．3．试剂配制································································· - 13 -2．2．2．鉴定方法······································································· - 14 -2．2．3．结果与讨论··································································· - 17 -2．3．本章小结····················································································· - 21 -第三章各假单胞菌株降油性能的初步研究············································· - 22 -3．1．材料····························································································· - 22 -3．1．1．菌株··············································································· - 22 -3．1．2．培养基··········································································· - 22 -3．2．方法····························································································· - 22 -3．3．结果与讨论················································································· - 23 -- 1 -山东轻工业学院2012届本科生毕业论文3．4．本章小结····················································································· - 25 -参考文献········································································································· - 25 -- 2 -山东轻工业学院2012届本科生毕业论文- 3 - 摘 要微生物采油是技术含量较高的一种提高采收率技术，包括微生物在油层中的生长、繁殖和代谢等生物化学过程，还包括微生物菌体、微生物营养液、微生物代谢产物在油层中的运移，以及与岩石、油、气、水的相互作用引起的岩石、油、气、水物性的改变。

两株石油降解菌的筛选及其生长特性DOI：10.14088/jki.issn0439-8114.2016.02.016土壤是人类赖以生存的自然基础和重要资源，近年来，随着社会经济发展对石油产品需求量增加，大量的石油及其加工品在勘探、开采、运输、加工及储存的过程中进入土壤，造成土壤的石油污染日趋严重，已成为世界性的环境问题[1，2]。

石油物理化学性质特殊，进入土壤后残留时间长，在自然条件下难以降解，给受污染土壤带来一系列的危害：石油黏性强，影响土壤的通透性，使土壤盐碱化、沥青化和板结化，破坏土壤结构和功能，在长期累积过程还会导致土壤中碳氮比和碳磷比失调，引起土壤微生物群落变化，破坏土壤微生物区系，影响植物生长；石油中所含的多环芳烃具有致癌、致突变、致畸变等作用，它们能通过食物链在动植物体内逐级富集，危及人类健康和生态安全[3，4]。

鉴于以上原因，修复和治理污染土壤已经成为迫切需要研究解决的问题。

目前石油污染土壤的修复方法主要有物理方法、化学方法和生物方法，其中微生物处理技术具有经济、有效和环境友好等特点，是目前修复石油污染土壤优先考虑的技术[5，6]。

微生物修复技术是微生物通过代谢作用将石油污染物转化为稳定、无毒的以二氧化碳、水和无机物为主的降解产物。

石油的降解程度和速度取决于微生物特性、污染物的性质和环境因素，而具有强降解能力的石油降解菌是决定生物修复效率的关键因素。

因此，需要有针对性地筛选适合本地区石油污染土壤的土著高效降解菌。

新疆是中国石油主产区，存在大面积石油污染土壤亟待修复[7，8]。

基于此，本研究从新疆石油污染土壤中筛选出2株高效石油降解菌，通过形态和生理生化试验对其进行了初步鉴定，并初步研究该菌的生长特性和培养条件，以期为新疆石油污染土壤的微生物修复提供试验基础。

1 材料与方法1.1 材料1.1.1 土壤和石油样品分离菌种土壤采自新疆油田区污染程度不同的位置，采样时取地表10 cm污染土壤1.0 kg，去除杂物，装入密封袋，于4 ℃冰箱保存。

石油污染土中微生物的分离鉴定及降解特性1. 引言1.1 研究背景石油污染是当前环境问题中的重要研究课题之一，随着石油开采和利用的增加，石油污染所造成的环境问题也日益严重。

石油是一种复杂的混合物，其中含有多种化合物，如烷烃、芳烃和环烷烃等，这些化合物对环境及生态系统产生了严重的影响。

石油污染土壤中的石油会对土壤中的微生物群落产生负面影响，破坏了土壤生态系统的平衡。

研究石油污染土中的微生物及其降解特性具有重要的意义。

微生物是地球上最古老的生物之一，具有多样性和适应性强的特点。

许多微生物能够利用石油中的有机物作为碳源和能量源，进行降解。

研究分离和鉴定石油降解菌，探究其生物降解特性和降解途径，对于解决石油污染问题具有重要意义。

本文旨在通过对石油污染土壤中微生物的分离、鉴定和降解特性研究，探讨微生物治理石油污染土壤的应用前景，为解决石油污染问题提供科学依据和参考。

1.2 研究目的研究的目的是通过对石油污染土中微生物的分离鉴定及降解特性的研究，探讨微生物在治理石油污染土壤中的作用机制，为有效治理石油污染提供科学依据。

具体目的包括：1.分离和鉴定石油降解菌，探究其生物降解机制和特性；2.研究石油降解菌的生物降解特性，探讨其在石油降解过程中的作用和影响因素；3.分析石油降解途径，揭示微生物在石油降解过程中的代谢途径和相关基因；4.探讨微生物治理石油污染土壤的应用前景，评估其在实际治理中的可行性和效果。

通过这些研究，可以揭示微生物在石油污染土壤中的重要性，评估其降解潜力，为未来的研究和治理提供科学依据和指导。

2. 正文2.1 石油污染土中微生物的分离和培养石油污染土中微生物的分离和培养是研究石油降解的重要步骤之一。

采集石油污染土样品，并将样品进行筛选和分离，获取微生物样品。

接着，将微生物进行培养，提供适当的营养物质和环境条件，促进微生物的生长和繁殖。

常用的培养基包括富含碳源和氮源的培养基，以及添加不同浓度石油类物质的培养基。

石油降解菌群的筛选、构建及其降解特性研究詹亚斌;张桥;陈凯伦;李方敏;马立安【期刊名称】《环境污染与防治》【年(卷),期】2017(039)008【摘要】为了构建高效石油降解混合菌群,从潜江某油田采集8个石油污染土样(分别记为S1~S8)和2个石油污染水样(分别记为W3、W4),以石油为唯一碳源进行富集驯化培养;采用外观评分、石油降解率、石油3组分降解率、饱和烃中正构烷烃色谱分析等方法筛选石油降解优势菌群,构建石油降解混合菌群;采用正交试验研究混合菌群最佳降解条件.结果表明,富集的10个石油降解菌群中S3、S4、S5、S6、S8为优势菌群,培养30 d后的石油降解率分别为21.67%、22.34%、27.23%、20.46%、19.99%;菌群W3、W4对石油乳化效果较好;混合菌群M3(S3+S4+S5+S8+W3+W4)为石油降解优势混合菌群,其最佳降解条件为35 ℃、pH 7.60、含油率1.70%,在最佳条件下培养30 d后,混合菌群M3对石油降解率达30.71%,比最优菌群S5的石油降解率提高了12.78%.【总页数】6页(P860-864,868)【作者】詹亚斌;张桥;陈凯伦;李方敏;马立安【作者单位】长江大学生命科学学院,湖北荆州 434025;长江大学生命科学学院,湖北荆州 434025;长江大学化学与环境工程学院,湖北荆州 434025;长江大学化学与环境工程学院,湖北荆州 434025;长江大学生命科学学院,湖北荆州 434025【正文语种】中文【相关文献】1.产生物表面活性剂石油降解菌的筛选及高效降解菌群的构建 [J], 夏铁骑;常慧萍;张建清2.石油降解菌的分离筛选及混合菌群的构建与优化 [J], 宁卓;刘雅慈;张胜;张翠云;何泽3.石油降解菌的筛选及复合菌群的构建 [J], 李乐;周飞;孙先锋4.石油污染土壤中石油降解菌的筛选及降解特性研究 [J], 汤小萌;马超;熊元林5.石油降解菌群的构建及其对混合烃的降解特性 [J], 范瑞娟;郭书海;李凤梅因版权原因，仅展示原文概要，查看原文内容请购买。

降解石油复合微生物菌剂的筛选研究摘要：从30个土样中筛选出3株高效原油降解菌株，它们为dch-16，dch-19和dch-20，7天后降油率分别为75.6%，80.3%和73.2%。

经鉴定，分别是脂肪酸芽孢杆菌属alicycolobacillus，芽孢肠状杆菌属sporomaculum和盐芽孢杆菌属halobacillus。

将此3株高效原油降解菌在原油培养基中进行复合实验，结果表明，在相同条件下复合菌降油效果优于单菌；菌株dch-19与dch-20复合的最佳原油降解条件为：接种量比1：1（总接种量为10%），ph值为7.5，底物浓度20mg·ml-1，温度35℃，原油降解时间为7天。

将实验复筛所得部分降油菌用于胜华炼厂废水处理，效果最好的是菌dch-19和dch-20的复合，处理两天后降油率达到80.2%。

表明复合菌株dch-19和dch-20有很强的适应能力。

abstract： we select three strains of high efficient oil degrading strains from 30 soil samples， i.e. the dch-16，dch-19 and dch-20. seven days later， the oil degradation rate turns to 75.6%， 80.3% and 73.2%. the three strains are identified as the fatty acid bacillus alicycolobacillus，bacillus sporomaculum and bacillus halobacillus. thereafter，we use the three strains of high efficient degradation bacteria for the crude oil compound experiments. the result shows that the compound bacteria has much better efficiency than single bacteria， and the optimized condition for dcd-19and dch-20 is that： the inoculation ratio gets 1：1 with total inoculation 10%， the ph value is 7.5， the concentration of substrate is 20mg. ml-1， and the temperature reaches 35℃，then the degradation of crude oil will be 7 days. further more，we introduce part of the microbial inoculants into the refinery wastewater treatment of shenghua refinery. the compound dch-19 and dch-20 shows the best performance of petroleum degradation which is up to 80.2%.关键词：筛选；石油降解；复合生物菌；炼厂废水key words： selection；petroleum degradation；complex microbial inoculants；refinery wastewater中图分类号：[p642.5] 文献标识码：a 文章编号：1006-4311（2013）13-0296-03————————————作者简介：徐宝刚（1979-），男，山东潍坊人，工程师，工学学士，环境工程给排水专业硕士在读。