柴油降解菌的筛选、降解条件优化及其降解机理研究
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摘要
石油烃降解菌广泛存在于自然界中。影响石油污染物降解效果的因素很多,如石油烃的种类和组成、物理状态、浓度、温度、pH值、供氧及营养物质等。石油烃的降解速率及最终降解效果是上述各种因素综合作用的结果。
由于石油烃的自然降解过程缓慢,而向污染环境中添加某些物质,优化降解条件,往往能加速石油污染物的生物降解过程。已有研究表明,表面活性物质可以促进石油烃的乳化,提高石油烃在水中溶解度及生物可利用性。与化学合成的表面活性剂相比,生物表面活性剂因其具有无毒、可生物降解、生态安全和高生物活性等特征越来越受到人们的关注。现在已经成为水体污染治理的热点。
本课题首先从长沙市某炼油厂受石油烃污染的土壤中分离筛选出以0#柴油为唯一碳源生长的9株石油烃降解菌。并对菌株的降解能力进行了研究,确定降解优势菌株。其中一株菌能在3d内能使柴油的降解率达到71.7%。经形态学特征和生理生化特性鉴定,这株菌为假单胞菌属。
其次本文研究了培养时间、pH值、接种量、温度、柴油浓度、N源对菌株降解率的影响,实验结果表明,该菌株在培养时间为6d、pH值为5.0、接种量为5%、温度为35℃、柴油浓度为1000mg/L、N源为(NH4)2SO4的条件下生长旺盛,降解率达到了80%。
此外本文还研究了如何从发酵液中提取表面活性剂,同时测定发酵液的表面张力,证明该菌能够产生生物表面活性剂,对产表面活性剂的条件进行优化,使表面活性剂达到最高产量。结果表明,假单胞菌在温度为30℃、初始pH为8.0～9.0、转速为200r/min、氮源为NH4NO3的条件下培养2d,培养液的表面张力可由69mN/m降至31.4mN/m,此时表面活性物质的产量达到了1.62g/L。将该菌产生的生物表面活性剂用于柴油降解实验,72h柴油去除率达到了85.7%,比不加表面活性剂的效果好。从降解前后柴油GC-MS图谱分析表明,该菌主要将C17-C29之间的直链烃和少量支链烃降解成C14-C18的短链烃。这为该菌株及其合成的表面活性物质应用于柴油污染土壤和水体的生物修复提供了有力的依据。
There are many oil degrading strains in the nature. The degradation of the petroleum hydrocarbons is influenced by many factors, such as the sort and composition of petroleum hydrocarbons, the physical state, the concentration, temperature, pH, oxygen supply, nutrients, et al. The degrade speed of petroleum hydrocarbons and the final effect is the interactional result of all the factors.
Because the natural degradation ratio of petroleum hydrocarbons is slow, adding some materials to the polluted environment, and optimizing the degradation condition can often accelerate the biodegradation of the petroleum pollutant. Some researches have indicated that surface-active compounds can promote the emulsification of the petroleum hydrocarbons, increase the solubility and the bioavailability of the petroleum hydrocarbons in the water. Compared with the chemical surfactants, biosurfactants are attentioned by more and more people because of their nontoxic, biodegradability, mildness to the environment, high bioactivity and so on. At present, it is becoming the hot of water pollution control. Our research was focus on the following aspects and received several results:
Firstly, nine diesel oil degradable strains were obtained from oil-contaminated soil by using screening cultural medium method which using 0# diesel oil as the sole carbon source. The degradation ratio of diesel oil by the strains was tested by using ultraviolet spectrometry method. One of the strains showed potential for further utilization with a degradation ratio of diesel oil by 71.7% incubating for 3 days. Based on the morphological, physiological and biochemical test results, the strain was identified as Pseudomonas sp..
Secondly, six factors that affect the strain growth and the degradation ratio to diesel oil, such as pH value of the culture medium, temperature, inoculation amount, N source, initial diesel oil concentration and incubation time, were also studied in the proposed paper. The result showed that the optimum culture condition for Pseudomonas sp. strain was pH5.0, inoculation by 5%, (NH4)2SO4 as the N source in the culture medium and incubation under 30℃. When the proposed strain grows under this condition, 80% of the diesel oil could be degraded with an initial concentration of 1000mg/L and incubating for 6d.
Furthermore, this paper also studied the extract of biosurfactant from broth. The Pseudomonas was capable of producing glycolipids biosurfactant by a series of tests including surface tension determination, thin-layer chromatography assay etc, and could reduce the surface tension of fermentation solution. The analysis of different culture conditions indicated that the biosurfactant produced by Pseudomonas was able to reduce the surface tension of fermentation solution from 69mN/m to 31.4mN/m when incubating for two days in conditions that the temperature was 30℃, the initial pH was 8.0～9.0, the shaking speed was 200r/min, the nitrogen source was NH4NO3. At the same time, the output of biosurfactant was 1.62g/L. Employing the biosurfactant to degrade diesel oil, 85.7% of the diesel oil was removed after 72h,it had good effect. Using the techniques of GC-MS, the effect of biosurfactant on degrading the composition of diesel oil was analyzed. The results showed it could degrade C17-C29 straight chain alkanes and some branched ones into C14-C18 short chain alkanes. This paper will provide strong basis for the strain and its biosurfactants applied in oil-contaminated soil and water.
由于石油烃的自然降解过程缓慢,而向污染环境中添加某些物质,优化降解条件,往往能加速石油污染物的生物降解过程。已有研究表明,表面活性物质可以促进石油烃的乳化,提高石油烃在水中溶解度及生物可利用性。与化学合成的表面活性剂相比,生物表面活性剂因其具有无毒、可生物降解、生态安全和高生物活性等特征越来越受到人们的关注。现在已经成为水体污染治理的热点。
本课题首先从长沙市某炼油厂受石油烃污染的土壤中分离筛选出以0#柴油为唯一碳源生长的9株石油烃降解菌。并对菌株的降解能力进行了研究,确定降解优势菌株。其中一株菌能在3d内能使柴油的降解率达到71.7%。经形态学特征和生理生化特性鉴定,这株菌为假单胞菌属。
其次本文研究了培养时间、pH值、接种量、温度、柴油浓度、N源对菌株降解率的影响,实验结果表明,该菌株在培养时间为6d、pH值为5.0、接种量为5%、温度为35℃、柴油浓度为1000mg/L、N源为(NH4)2SO4的条件下生长旺盛,降解率达到了80%。
此外本文还研究了如何从发酵液中提取表面活性剂,同时测定发酵液的表面张力,证明该菌能够产生生物表面活性剂,对产表面活性剂的条件进行优化,使表面活性剂达到最高产量。结果表明,假单胞菌在温度为30℃、初始pH为8.0～9.0、转速为200r/min、氮源为NH4NO3的条件下培养2d,培养液的表面张力可由69mN/m降至31.4mN/m,此时表面活性物质的产量达到了1.62g/L。将该菌产生的生物表面活性剂用于柴油降解实验,72h柴油去除率达到了85.7%,比不加表面活性剂的效果好。从降解前后柴油GC-MS图谱分析表明,该菌主要将C17-C29之间的直链烃和少量支链烃降解成C14-C18的短链烃。这为该菌株及其合成的表面活性物质应用于柴油污染土壤和水体的生物修复提供了有力的依据。
There are many oil degrading strains in the nature. The degradation of the petroleum hydrocarbons is influenced by many factors, such as the sort and composition of petroleum hydrocarbons, the physical state, the concentration, temperature, pH, oxygen supply, nutrients, et al. The degrade speed of petroleum hydrocarbons and the final effect is the interactional result of all the factors.
Because the natural degradation ratio of petroleum hydrocarbons is slow, adding some materials to the polluted environment, and optimizing the degradation condition can often accelerate the biodegradation of the petroleum pollutant. Some researches have indicated that surface-active compounds can promote the emulsification of the petroleum hydrocarbons, increase the solubility and the bioavailability of the petroleum hydrocarbons in the water. Compared with the chemical surfactants, biosurfactants are attentioned by more and more people because of their nontoxic, biodegradability, mildness to the environment, high bioactivity and so on. At present, it is becoming the hot of water pollution control. Our research was focus on the following aspects and received several results:
Firstly, nine diesel oil degradable strains were obtained from oil-contaminated soil by using screening cultural medium method which using 0# diesel oil as the sole carbon source. The degradation ratio of diesel oil by the strains was tested by using ultraviolet spectrometry method. One of the strains showed potential for further utilization with a degradation ratio of diesel oil by 71.7% incubating for 3 days. Based on the morphological, physiological and biochemical test results, the strain was identified as Pseudomonas sp..
Secondly, six factors that affect the strain growth and the degradation ratio to diesel oil, such as pH value of the culture medium, temperature, inoculation amount, N source, initial diesel oil concentration and incubation time, were also studied in the proposed paper. The result showed that the optimum culture condition for Pseudomonas sp. strain was pH5.0, inoculation by 5%, (NH4)2SO4 as the N source in the culture medium and incubation under 30℃. When the proposed strain grows under this condition, 80% of the diesel oil could be degraded with an initial concentration of 1000mg/L and incubating for 6d.
Furthermore, this paper also studied the extract of biosurfactant from broth. The Pseudomonas was capable of producing glycolipids biosurfactant by a series of tests including surface tension determination, thin-layer chromatography assay etc, and could reduce the surface tension of fermentation solution. The analysis of different culture conditions indicated that the biosurfactant produced by Pseudomonas was able to reduce the surface tension of fermentation solution from 69mN/m to 31.4mN/m when incubating for two days in conditions that the temperature was 30℃, the initial pH was 8.0～9.0, the shaking speed was 200r/min, the nitrogen source was NH4NO3. At the same time, the output of biosurfactant was 1.62g/L. Employing the biosurfactant to degrade diesel oil, 85.7% of the diesel oil was removed after 72h,it had good effect. Using the techniques of GC-MS, the effect of biosurfactant on degrading the composition of diesel oil was analyzed. The results showed it could degrade C17-C29 straight chain alkanes and some branched ones into C14-C18 short chain alkanes. This paper will provide strong basis for the strain and its biosurfactants applied in oil-contaminated soil and water.
引文
[1] Jonathan D V H,Ajay S, Owen P W.Recent Advances in Petroleum Microbiology[J].Microbiology and Molecular Biology Reviews,2003,67(4):503-549
[2]王家玲.环境微生物.北京:高等教育出版社,1998,55-78
[3]张从,夏立江.污染土壤的生物修复技术.北京:中国环境出版社,2000,116-145
[4]李宝明.石油降解细菌的分离与筛选研究:[沈阳农业大学硕士学位论文].沈阳:沈阳农业大学环境工程系,2004,3-18
[5]曾国涛,徐梦虹.石油地球化学.北京:石油工业出版社,1990,19-54
[6]吕志萍,程龙飞.石油污染土壤中石油含量对玉米的影响.油气田环境保护, 2001,11(1):36-37
[7]丁克强,孙铁衍,李培军.石油污染土壤的生物修复技术.生态学杂志, 2000,19(2):50-55
[8] Truax D D,Brittu R,Sherrard J H.Bench-scalt studies of reactor based treatment of fuel-contaminated soils[J].Waste Management,1995,15(5-6):351-357
[9]李凯峰,温青.石油污染土壤的生物处理技术[J].应用科技,2002,29(10):62-64
[10]姚德明,许华夏,张海荣,等.石油污染土壤生物修复过程中微生物生态研究[J].生态学杂志,2002,21(1):26-28
[11] Herbes,Schwall S R.Microbial transformation of polycyclicaromatic hydrocarbons in pristine and petroleum-contaminated sediments.Applied and Environmental Microbiology[J],1978,20(3):306-316
[12]何良菊,魏德洲,张维庆,等.土壤微生物处理石油污染的研究[M].环境科学进展,1999,7(3):110-115
[13]李慧.石油烃污染对稻田土壤微生物生态系统的影响:[中科院博士学位论文].沈阳:中国科学院沈阳应用生态研究所,2005,5-18
[14]解岳.延河流域石油类污染物非点源污染特征及其在客流沉积物中的吸附与释放:[西安建筑大学博士学位论文].西安:西安建筑科技大学,1999:13-21
[15]朱孟龙.石油成品油对健康的危害及防护[J].石油库与加油站,1999,8(3):39-40
[16] Haimi J.Decomposer animals and bioremediation of soils[J].Environ.Poll,2000,107(3):233-238
[17]陈坚,华兆哲.生物表面活性剂在环境工程中的应用[J].环境科学,1996,17 (4): 84-87
[18] Wehner H,Teschner M,K.Bosecker.Org.Gechem[M].New York:ChemicalPublishion,1986:463-471
[19] Milner C W,Rogers M A,Evans C R.Petroleum transformations inreservoir[J]. Geochem Explor,1977,7(10):101-153
[20] Seifer W K,Moldowan J M.The effect of biodegradation on steanes and terpanes in crude oils[J].Geochim cosmo chim Acta,1979,43(4):111-126
[21] Mahlon C k.The effect of degradation on crude oil bulk and molecular composition[J].Oil and Chemical Pollution,1988,11(2):89-112
[22] Scherrer P,Mille G.Marine Pollution Bulletin[M].London:Industry Publishion,1989:430-432
[23] Dkpokwasili G C,Ddokuma L O.Effect of salinity on biodegradation of oil spill dispersants.Waste management[J],1990,10(1):141-146
[24]常志州,何加骏,Weaver R W.两种土壤上接种微生物对提高石油降解率的影响[J].农业环境保护,1998,17(1):15-17
[25]常志州,何加骏,Weaver R W.水势与用氮量对土壤中石油降解的影响[J].农村生态环境,1997,13(1):61-65
[26]陈碧娥,郭厚宝,苏荣西,等.湄洲湾天然菌群对石油烃的降解作用[J].华侨大学学报,2000,21(2):182-186.
[27]陈碧娥,刘祖同.湄洲湾海洋细菌降解石油烃研究[J].石油学报,2001,17(5): 31-35.
[28]丁克强,骆永明.多环芳烃污染土壤的生物修复[J].土壤,2001,33(4):169-178
[29]龚利萍,张甲耀,罗宇煊.土壤微生物降解石油污染物[J].上海环境科学,2001,20 (4):201-202
[30]巩宗强,李培军,郭书海,等.多环芳烃污染土壤的生物泥浆法修复[J].环境科学,2001,22(5):112-116
[31]姜昌亮,孙铁衍,李培军,等.石油污染土壤长料堆式异位生物修复技术研究[J].应用生态学报,2001,12(2):279-282
[32]谢明杰,谢正等.微生物降解原油提高原油采油率的研究[J].抚顺石油学院学报, 1999,19(2):29-32
[33]林凤翱,于占国,李洪,等.海洋丝状真菌降解原油研究[J].海洋学报,1997,19(6): 68-75
[34]陆光华,万蕾,苏瑞莲,等.石油烃类污染土壤的生物修复技术研究进展[J],生态环境,2003,12(2):220-223
[35]魏德洲,秦熠民.H2O2在石油污染土壤微生物治理过程中的作用[J].中国环境科学,1997,17(5):429-432
[36] Ellis B,Harold P,Kronberg H.Bioremediation of a creosote contaminatedsite[M].Environ Technol,1991,12(1):447-459
[37]张文娟,沈德中,张从,等.堆制处理过程中的多环芳烃降解[J].应用与环境生物学报,1999,22(6):605-609
[38] Castldi F J,Ford D L.Bioremediation of petrochemical water sludges[J].Water Science and Technology,1992,25(3):207-212
[39]李法云.污染土壤生物修复理论基础与技术[M].北京:化学工业出版社,2006,42-50
[40] Higgings L J.Microb.Ecosyst Proc[M].Chicago:Oil Press.1978:80-84
[41]潘学芳,烃类降解菌的筛选及其在微生物技术处理石油污染物中的应用:[山东大学硕士学位论文].济南:山东大学,2001:32-38
[42] MeKenna E.Biochim.Biopbys[J].Annual Review of Microbiol.1965,19(1):183-187
[43] Andrew W,Jackson.Potential for Enhancement of Biodegradation of Crude Oil in Louisiana Salt Marshes Nutrient Amendments.Water[J].Air and Soil Pollution,1999,109(5):343-355
[44] Cemiglia C E,Heitkamp M A.Microbialde gradation of PAHs in the aquatic environment[J].Environmental Research Laboratory,1988,20(3):34-38
[45]华兆哲,陈坚,朱文昌,等.假丝酵母(Candida antartic WSH 112)生产生物表面活性剂及其降解正构烷烃的研究[J].南京大学学报,1998,34(2):149-154
[46] Li K Y,Annamali S N,Hopper J R. Rate controlling model for bioremediation of oil contaminated soil[J].Environmental Progress,1993,12(4):257-261
[47] Carriere P P E,Mesania F A.Enhanced biodegradation of creosote-contaminated Soil[J].Waste Management,1995,15(8):579-583
[48] Dibble J T,Bartha R.Effect of environmental parameters on the biodegradation of oil sludge[J].J Appl Environ Microbial.1979,37(5):729-739
[49] Brink R H.Biodegradation of organic chemicals in the environment[M].London:Enviroomental health chemistry Technomic Pubishing Company,1981:1-10
[50] Edward J C,Paul T K,et al.Principles and practices for Petroleum contaminated Soils[M].Berlin:Lewis Publishers INC,1993:111-129
[51]金文标,宋莉晖,等.油污土壤微生物治理的影响[j].环境保护,1998,10(1):27-28
[52]郑远扬.石油污染生化治理的进展.国外环境科学技术[J],1993,20(3):46-59
[53] Atlas R M.Microbial hydrocarbon degradation-biodegradation of oil spills[J]. Chem Tech Bio-technol,1991,52(2):149-156
[54]金文标.盐度对油污土壤生物治理的影响研究[J].钻采工艺,1998,21(4):71-72
[55] Rhykerd R L.Influence of salinity on bioremediation of oil in soil[J].Environ Pollute,1995,90(1):127-130
[56] J.M.亨特著,石油地球化学和地质学[M].胡伯良.第一版.北京:石油工业出版社, 1986:286-280
[57]吴玉新.紫外分光光度法测定污水中油含量的研究[J].石化技术,1998,5(2): 112-114
[58]黄红,吴红雨,朱红昆.紫外分光光度法测定污水中油的分析方法探讨[J].黑龙江环境通报,2000,24(3):64-96
[59]刘汉初,胡玉.水中油的测定方法探讨[J].环境监测管理与技术,1997,9(3):38-39
[60] Robert M,Garrett.Photo oxidation of Crude Oils[M].Environ Sci Technol.1998,32(2):3719-3723
[61] Rasiah V.Biodegradation of an Oily Waste as Influenced By Nitrogen Forms and Sources[J].Water,Air and Soil Pollution,1991,65(4):143-151
[62] Jackson W A.Potential for Enhancement of Biodegradation of Crude Oil in Louisiana Salt Marshes Using Nutrient Amendments[J].Water,Air and Soil Pollution,1999,10(9):343-355
[63]庄毅,马梦瑞.生物技术与生物表面活性剂[J].山东化工,1994,22(1):20-26
[64]肖进新,赵振国.表面活性剂应用原理[M].北京:化学工业出版社,2003,395-408
[65]马歌丽,彭新榜,马翠卿,等.生物表面活性剂及其应用[J].中国生物工程杂志, 2003,23(5):42-45
[66]徐志伟,尤勤.生物表面活性剂的工业应用[J].生物技术,1995,5(3):6-8
[67]石梅,孙凤荣,侯兆伟,等.生物表面活性剂在油田中的应用[J].日用化学品科学,2000,23(1):176-179
[68]赵淑梅,郑西来,高增文,等.生物表面活性剂及其在油污染生物修复技术中的应用[J].海洋科学进展,2005,23(2):234-238
[69] Desai J D,Banat I M.Microbial production of surfactants and their commercial potential.Microbial.Molec[J].Biol.Rev,1997,61(1):47-64
[70]宋玉芳,孙铁衍,许华夏.表面活性剂TW-80对土壤中多环芳烃生物降解的影响[J].应用生态学报,1999,10(2):103-105
[71]朱万琴,王淑华,冯健,等.S-1菌株的筛选及乳化剂在其降解石油过程中的促进作用初探[J].微生物学杂志,1997,17(1):40-42
[72]张小凡.多环芳烃化合物菲分解菌的分离鉴定及分解特性研究[J].上海环境科学,2003,22(8):544-547
[73]张翠竹,张心平,梁凤来,等.一株地衣芽饱杆菌产生的生物表面活性剂[J].南开大学学报,2000,33(4):41-46
[74] Hayes M E,Nestaas E,Hrebenar K R,et al.Microbial surfactants[J].Chen tech,1986,22(4):239-243
[75] Banat I M,Samarah N,Murad M, et al.Biosurfactants production and use in oil tank clear-up[J].World J Microbiol Biotechnol,1991,32(7):80-84
[76]李道山,廖广志,杨林.生物表面活性剂作为牺牲剂在三元复合驱中的应用研究.石油勘探与开[J].微生物学通报,2002,29(2):106-109
[77] Shephord R,ROCKEY J,Shutherland I W,et al.Novel bioemulsifier from microorganisms for use in foods[J]. J Biotechnol,1995,40(3):207-217
[78] Inoue S,Kimwa Y,Kinta M, et al.German Patent 2905252 to Kao Soap CO[M].Tokyo:Labrotory,1997:110-120
[79] White R T,Damm D,Miller J,et al.Isolation and characterization of the human pulmonary surfactant apoprotein gene[J].Nature,1985,31(7):316-320
[80] Stanghellini M E,Miller R M.Biosurfactants: Their identity and potential efficacy in the biological control of zoosporic plant pathogens[J].Plant Dis,1997,8(1):4-12
[81] Hafeburg D,Hommel R,Kleber H P,et al.Antiphytovirale aktivitat von rhamnolipid aus Pseudominasa eruginosa acta[J].Biotechnol,1987,20(7):353-356
[82] Awashti N,KumarA,Makkar R S,et al.Enhanced biodegradation of endosulfan,a chlorinated pesticide in presence of a biosurfactant[J].Environ Sci Health, 1999,34(6):234-239
[83]江开治.微生物表面活性剂在工业、农业、医药卫生和环境保护中的应用[J].生物技术通报,2002,22(3):45-49
[85] Bossert I,Bartba R.Plant growth in soil with history of oily sludge disposal[J].Soil science,1985,14(1):75-77
[85] Liu D.Fate of petroleum hydrocarbons in sewage sludge after land disposal[J].Bull Environ Contam toxicol,1980,25(3):612-622
[86]陈玉成.土壤污染的生物修复[J].环境科学动态,1999,12(5):7-11
[87] Aries E,Doumenq P,Artaud J,et al.Effects of petroleum hydrocarbons on the phospholipid fatty acid composition of a consortium composed of marine hydrocarbon-degrading bacteria[J].Organic Geochem,2001,32(7):891-903
[88]周林红,吴燕.紫外分光光度法测定炼油废水中的石油类含量[J].石化技术与应用, 2004,22(6):456-458
[89]东秀珠,蔡妙英,等.常见细菌系统鉴定手册[M].北京:科学出版社. 2001:100-113
[90]袁红莉,杨金水,王占生,等.降解石油微生物菌种的筛选及降解特性[J].中国环境科学,2003,23(2):157-161
[91] Rahman K S M,Vasudevan N,Lakshmanaperumalsamy P Enhancement of biosurfactant production to emulsity different hydrocarbons[J].Environ Pollut,1999,34(6):87-93
[92] Salminen J M,Tuomi P M,Suortti A M,et al.Potential for aerobic and anaerobic biodegradation of petroleum hydrocarbons in boreal subsurface[J].Biodegradation,2004,15(1):29-39
[93] Leathy J G,Colwell R R.Microbial degradation of hydrocarbons in environment[J].Microbiol Rev,1990,54(3):305-315
[94] Rambeloarisoa E,Rontani J F,Giusti G,et al.Degradation of crude oil by a mixed population of bacteria isolated from sea surface foams[J].Marine Biol,1984,83(5):69-81
[95]陆泗进,王红旗.汽油降解菌的分离及降解研究[M].环境科学研究,2006,19(4: 95-99
[96]何良菊,李培杰,魏德洲,等.石油烃微生物降解的营养平衡及降解机理[M].环境科学,2004,25(1):91-94
[97] Carriere P E P.Enhanced biodegradation of creosote-contaminated soil[J].Waste Management,1995,33(4):579-583
[98] Kosaric N.Biosurfactants Production, Properties and Applications[M].NewYork: Marcel Dekker Inc,1993:66-97
[99] SARNEY D B,VULFEN E N.Application of enzymes to the synthesis of surfactants[J].Trends Biotechnol,1995,13(2):164-172
[100]牛明芬,李凤梅,韩晓日,等.生物表面活性剂产生菌的筛选及表面活性剂稳定性研究[M].生态学杂志,2005,24(6):631-634
[101]王修垣,田新玉,宋沛然.菌株H255原油发酵液乳化物质的分析[M].微生物学报,1982,22(3):269-275
[102] Harvey S,Elashvili I,Valdes JJ,et al.Enhanced removel of Exxon Valdez spilled oil from Alaskan gravel by a microbial surfactant[J].Biotechnology,1990,8(3):228-230
[103] Whang L M,Liu P G,Ma C C,et al.Application of biosurfactants, rhamnolipid and surfactin for enhanced biodegradation of diesel-contaminated water and soil[J].Journal of Hazardous Materials,2007,25(5):63-71
[104]傅海燕,曾光明,袁兴中,等.生物表面活性剂的分离提纯及其发展前景[J].生物学杂志,2003,20(6):1-4
[105]卢国满.产表面活性剂菌株的筛选、发酵条件优化及定量研究:[湖南大学硕士学位论文].长沙:湖南大学环境科学与工程学院,2006:8-10
[106] Cooper DG,Macdonald CR,Duff SBJ.Enhanced production of surfactin from Bacillus subtilis by continuous product removel and metal cation additions[J].Appl Environ Microbiol,1981,42(3):408-412
[107]麦有斌,尹华,叶锦韶,等.假单胞菌表面活性物主产生与性能[J].环境科学与技术,2006,29(2):12-14
[108] Velanker S K,Barneet S M,Houston C W,et al.Microbiol growth on hydrocarbons-some experimental results[J].Biotechnol Bioeng,1975,17(2): 241-251
[109] Davis D A,Lynch H C,Varley J.The production of surfactin in batch culture by Bacillus subtilis 21332 is strongly influenced by the conditions of nitrogen metabolism[J].Enzyme and Microbial Technol,1999,25(3):322-329
[110] Olness A,Clapp C G.Microbial degradation of montmorillo-nite-dextran complex[J].Soil Sci Soc AM Pro,1972,37(5):37-45
[111] Sen R,Swaminathan T.Application of response surface methodology to evaluate the optimum environmental conditions for the enhanced production of surfactin[J].Apply of Micrology Biotechnology, 997,47(8):358-363
[112] Fu H Y,Zeng G M,Huang G H,et al.Isolation of biosurfactant-producing bacteria from compost and their prospective application in composting[J].Transaction of Nonferrous Metals Societyof China,2004,14(1):131-134
[2]王家玲.环境微生物.北京:高等教育出版社,1998,55-78
[3]张从,夏立江.污染土壤的生物修复技术.北京:中国环境出版社,2000,116-145
[4]李宝明.石油降解细菌的分离与筛选研究:[沈阳农业大学硕士学位论文].沈阳:沈阳农业大学环境工程系,2004,3-18
[5]曾国涛,徐梦虹.石油地球化学.北京:石油工业出版社,1990,19-54
[6]吕志萍,程龙飞.石油污染土壤中石油含量对玉米的影响.油气田环境保护, 2001,11(1):36-37
[7]丁克强,孙铁衍,李培军.石油污染土壤的生物修复技术.生态学杂志, 2000,19(2):50-55
[8] Truax D D,Brittu R,Sherrard J H.Bench-scalt studies of reactor based treatment of fuel-contaminated soils[J].Waste Management,1995,15(5-6):351-357
[9]李凯峰,温青.石油污染土壤的生物处理技术[J].应用科技,2002,29(10):62-64
[10]姚德明,许华夏,张海荣,等.石油污染土壤生物修复过程中微生物生态研究[J].生态学杂志,2002,21(1):26-28
[11] Herbes,Schwall S R.Microbial transformation of polycyclicaromatic hydrocarbons in pristine and petroleum-contaminated sediments.Applied and Environmental Microbiology[J],1978,20(3):306-316
[12]何良菊,魏德洲,张维庆,等.土壤微生物处理石油污染的研究[M].环境科学进展,1999,7(3):110-115
[13]李慧.石油烃污染对稻田土壤微生物生态系统的影响:[中科院博士学位论文].沈阳:中国科学院沈阳应用生态研究所,2005,5-18
[14]解岳.延河流域石油类污染物非点源污染特征及其在客流沉积物中的吸附与释放:[西安建筑大学博士学位论文].西安:西安建筑科技大学,1999:13-21
[15]朱孟龙.石油成品油对健康的危害及防护[J].石油库与加油站,1999,8(3):39-40
[16] Haimi J.Decomposer animals and bioremediation of soils[J].Environ.Poll,2000,107(3):233-238
[17]陈坚,华兆哲.生物表面活性剂在环境工程中的应用[J].环境科学,1996,17 (4): 84-87
[18] Wehner H,Teschner M,K.Bosecker.Org.Gechem[M].New York:ChemicalPublishion,1986:463-471
[19] Milner C W,Rogers M A,Evans C R.Petroleum transformations inreservoir[J]. Geochem Explor,1977,7(10):101-153
[20] Seifer W K,Moldowan J M.The effect of biodegradation on steanes and terpanes in crude oils[J].Geochim cosmo chim Acta,1979,43(4):111-126
[21] Mahlon C k.The effect of degradation on crude oil bulk and molecular composition[J].Oil and Chemical Pollution,1988,11(2):89-112
[22] Scherrer P,Mille G.Marine Pollution Bulletin[M].London:Industry Publishion,1989:430-432
[23] Dkpokwasili G C,Ddokuma L O.Effect of salinity on biodegradation of oil spill dispersants.Waste management[J],1990,10(1):141-146
[24]常志州,何加骏,Weaver R W.两种土壤上接种微生物对提高石油降解率的影响[J].农业环境保护,1998,17(1):15-17
[25]常志州,何加骏,Weaver R W.水势与用氮量对土壤中石油降解的影响[J].农村生态环境,1997,13(1):61-65
[26]陈碧娥,郭厚宝,苏荣西,等.湄洲湾天然菌群对石油烃的降解作用[J].华侨大学学报,2000,21(2):182-186.
[27]陈碧娥,刘祖同.湄洲湾海洋细菌降解石油烃研究[J].石油学报,2001,17(5): 31-35.
[28]丁克强,骆永明.多环芳烃污染土壤的生物修复[J].土壤,2001,33(4):169-178
[29]龚利萍,张甲耀,罗宇煊.土壤微生物降解石油污染物[J].上海环境科学,2001,20 (4):201-202
[30]巩宗强,李培军,郭书海,等.多环芳烃污染土壤的生物泥浆法修复[J].环境科学,2001,22(5):112-116
[31]姜昌亮,孙铁衍,李培军,等.石油污染土壤长料堆式异位生物修复技术研究[J].应用生态学报,2001,12(2):279-282
[32]谢明杰,谢正等.微生物降解原油提高原油采油率的研究[J].抚顺石油学院学报, 1999,19(2):29-32
[33]林凤翱,于占国,李洪,等.海洋丝状真菌降解原油研究[J].海洋学报,1997,19(6): 68-75
[34]陆光华,万蕾,苏瑞莲,等.石油烃类污染土壤的生物修复技术研究进展[J],生态环境,2003,12(2):220-223
[35]魏德洲,秦熠民.H2O2在石油污染土壤微生物治理过程中的作用[J].中国环境科学,1997,17(5):429-432
[36] Ellis B,Harold P,Kronberg H.Bioremediation of a creosote contaminatedsite[M].Environ Technol,1991,12(1):447-459
[37]张文娟,沈德中,张从,等.堆制处理过程中的多环芳烃降解[J].应用与环境生物学报,1999,22(6):605-609
[38] Castldi F J,Ford D L.Bioremediation of petrochemical water sludges[J].Water Science and Technology,1992,25(3):207-212
[39]李法云.污染土壤生物修复理论基础与技术[M].北京:化学工业出版社,2006,42-50
[40] Higgings L J.Microb.Ecosyst Proc[M].Chicago:Oil Press.1978:80-84
[41]潘学芳,烃类降解菌的筛选及其在微生物技术处理石油污染物中的应用:[山东大学硕士学位论文].济南:山东大学,2001:32-38
[42] MeKenna E.Biochim.Biopbys[J].Annual Review of Microbiol.1965,19(1):183-187
[43] Andrew W,Jackson.Potential for Enhancement of Biodegradation of Crude Oil in Louisiana Salt Marshes Nutrient Amendments.Water[J].Air and Soil Pollution,1999,109(5):343-355
[44] Cemiglia C E,Heitkamp M A.Microbialde gradation of PAHs in the aquatic environment[J].Environmental Research Laboratory,1988,20(3):34-38
[45]华兆哲,陈坚,朱文昌,等.假丝酵母(Candida antartic WSH 112)生产生物表面活性剂及其降解正构烷烃的研究[J].南京大学学报,1998,34(2):149-154
[46] Li K Y,Annamali S N,Hopper J R. Rate controlling model for bioremediation of oil contaminated soil[J].Environmental Progress,1993,12(4):257-261
[47] Carriere P P E,Mesania F A.Enhanced biodegradation of creosote-contaminated Soil[J].Waste Management,1995,15(8):579-583
[48] Dibble J T,Bartha R.Effect of environmental parameters on the biodegradation of oil sludge[J].J Appl Environ Microbial.1979,37(5):729-739
[49] Brink R H.Biodegradation of organic chemicals in the environment[M].London:Enviroomental health chemistry Technomic Pubishing Company,1981:1-10
[50] Edward J C,Paul T K,et al.Principles and practices for Petroleum contaminated Soils[M].Berlin:Lewis Publishers INC,1993:111-129
[51]金文标,宋莉晖,等.油污土壤微生物治理的影响[j].环境保护,1998,10(1):27-28
[52]郑远扬.石油污染生化治理的进展.国外环境科学技术[J],1993,20(3):46-59
[53] Atlas R M.Microbial hydrocarbon degradation-biodegradation of oil spills[J]. Chem Tech Bio-technol,1991,52(2):149-156
[54]金文标.盐度对油污土壤生物治理的影响研究[J].钻采工艺,1998,21(4):71-72
[55] Rhykerd R L.Influence of salinity on bioremediation of oil in soil[J].Environ Pollute,1995,90(1):127-130
[56] J.M.亨特著,石油地球化学和地质学[M].胡伯良.第一版.北京:石油工业出版社, 1986:286-280
[57]吴玉新.紫外分光光度法测定污水中油含量的研究[J].石化技术,1998,5(2): 112-114
[58]黄红,吴红雨,朱红昆.紫外分光光度法测定污水中油的分析方法探讨[J].黑龙江环境通报,2000,24(3):64-96
[59]刘汉初,胡玉.水中油的测定方法探讨[J].环境监测管理与技术,1997,9(3):38-39
[60] Robert M,Garrett.Photo oxidation of Crude Oils[M].Environ Sci Technol.1998,32(2):3719-3723
[61] Rasiah V.Biodegradation of an Oily Waste as Influenced By Nitrogen Forms and Sources[J].Water,Air and Soil Pollution,1991,65(4):143-151
[62] Jackson W A.Potential for Enhancement of Biodegradation of Crude Oil in Louisiana Salt Marshes Using Nutrient Amendments[J].Water,Air and Soil Pollution,1999,10(9):343-355
[63]庄毅,马梦瑞.生物技术与生物表面活性剂[J].山东化工,1994,22(1):20-26
[64]肖进新,赵振国.表面活性剂应用原理[M].北京:化学工业出版社,2003,395-408
[65]马歌丽,彭新榜,马翠卿,等.生物表面活性剂及其应用[J].中国生物工程杂志, 2003,23(5):42-45
[66]徐志伟,尤勤.生物表面活性剂的工业应用[J].生物技术,1995,5(3):6-8
[67]石梅,孙凤荣,侯兆伟,等.生物表面活性剂在油田中的应用[J].日用化学品科学,2000,23(1):176-179
[68]赵淑梅,郑西来,高增文,等.生物表面活性剂及其在油污染生物修复技术中的应用[J].海洋科学进展,2005,23(2):234-238
[69] Desai J D,Banat I M.Microbial production of surfactants and their commercial potential.Microbial.Molec[J].Biol.Rev,1997,61(1):47-64
[70]宋玉芳,孙铁衍,许华夏.表面活性剂TW-80对土壤中多环芳烃生物降解的影响[J].应用生态学报,1999,10(2):103-105
[71]朱万琴,王淑华,冯健,等.S-1菌株的筛选及乳化剂在其降解石油过程中的促进作用初探[J].微生物学杂志,1997,17(1):40-42
[72]张小凡.多环芳烃化合物菲分解菌的分离鉴定及分解特性研究[J].上海环境科学,2003,22(8):544-547
[73]张翠竹,张心平,梁凤来,等.一株地衣芽饱杆菌产生的生物表面活性剂[J].南开大学学报,2000,33(4):41-46
[74] Hayes M E,Nestaas E,Hrebenar K R,et al.Microbial surfactants[J].Chen tech,1986,22(4):239-243
[75] Banat I M,Samarah N,Murad M, et al.Biosurfactants production and use in oil tank clear-up[J].World J Microbiol Biotechnol,1991,32(7):80-84
[76]李道山,廖广志,杨林.生物表面活性剂作为牺牲剂在三元复合驱中的应用研究.石油勘探与开[J].微生物学通报,2002,29(2):106-109
[77] Shephord R,ROCKEY J,Shutherland I W,et al.Novel bioemulsifier from microorganisms for use in foods[J]. J Biotechnol,1995,40(3):207-217
[78] Inoue S,Kimwa Y,Kinta M, et al.German Patent 2905252 to Kao Soap CO[M].Tokyo:Labrotory,1997:110-120
[79] White R T,Damm D,Miller J,et al.Isolation and characterization of the human pulmonary surfactant apoprotein gene[J].Nature,1985,31(7):316-320
[80] Stanghellini M E,Miller R M.Biosurfactants: Their identity and potential efficacy in the biological control of zoosporic plant pathogens[J].Plant Dis,1997,8(1):4-12
[81] Hafeburg D,Hommel R,Kleber H P,et al.Antiphytovirale aktivitat von rhamnolipid aus Pseudominasa eruginosa acta[J].Biotechnol,1987,20(7):353-356
[82] Awashti N,KumarA,Makkar R S,et al.Enhanced biodegradation of endosulfan,a chlorinated pesticide in presence of a biosurfactant[J].Environ Sci Health, 1999,34(6):234-239
[83]江开治.微生物表面活性剂在工业、农业、医药卫生和环境保护中的应用[J].生物技术通报,2002,22(3):45-49
[85] Bossert I,Bartba R.Plant growth in soil with history of oily sludge disposal[J].Soil science,1985,14(1):75-77
[85] Liu D.Fate of petroleum hydrocarbons in sewage sludge after land disposal[J].Bull Environ Contam toxicol,1980,25(3):612-622
[86]陈玉成.土壤污染的生物修复[J].环境科学动态,1999,12(5):7-11
[87] Aries E,Doumenq P,Artaud J,et al.Effects of petroleum hydrocarbons on the phospholipid fatty acid composition of a consortium composed of marine hydrocarbon-degrading bacteria[J].Organic Geochem,2001,32(7):891-903
[88]周林红,吴燕.紫外分光光度法测定炼油废水中的石油类含量[J].石化技术与应用, 2004,22(6):456-458
[89]东秀珠,蔡妙英,等.常见细菌系统鉴定手册[M].北京:科学出版社. 2001:100-113
[90]袁红莉,杨金水,王占生,等.降解石油微生物菌种的筛选及降解特性[J].中国环境科学,2003,23(2):157-161
[91] Rahman K S M,Vasudevan N,Lakshmanaperumalsamy P Enhancement of biosurfactant production to emulsity different hydrocarbons[J].Environ Pollut,1999,34(6):87-93
[92] Salminen J M,Tuomi P M,Suortti A M,et al.Potential for aerobic and anaerobic biodegradation of petroleum hydrocarbons in boreal subsurface[J].Biodegradation,2004,15(1):29-39
[93] Leathy J G,Colwell R R.Microbial degradation of hydrocarbons in environment[J].Microbiol Rev,1990,54(3):305-315
[94] Rambeloarisoa E,Rontani J F,Giusti G,et al.Degradation of crude oil by a mixed population of bacteria isolated from sea surface foams[J].Marine Biol,1984,83(5):69-81
[95]陆泗进,王红旗.汽油降解菌的分离及降解研究[M].环境科学研究,2006,19(4: 95-99
[96]何良菊,李培杰,魏德洲,等.石油烃微生物降解的营养平衡及降解机理[M].环境科学,2004,25(1):91-94
[97] Carriere P E P.Enhanced biodegradation of creosote-contaminated soil[J].Waste Management,1995,33(4):579-583
[98] Kosaric N.Biosurfactants Production, Properties and Applications[M].NewYork: Marcel Dekker Inc,1993:66-97
[99] SARNEY D B,VULFEN E N.Application of enzymes to the synthesis of surfactants[J].Trends Biotechnol,1995,13(2):164-172
[100]牛明芬,李凤梅,韩晓日,等.生物表面活性剂产生菌的筛选及表面活性剂稳定性研究[M].生态学杂志,2005,24(6):631-634
[101]王修垣,田新玉,宋沛然.菌株H255原油发酵液乳化物质的分析[M].微生物学报,1982,22(3):269-275
[102] Harvey S,Elashvili I,Valdes JJ,et al.Enhanced removel of Exxon Valdez spilled oil from Alaskan gravel by a microbial surfactant[J].Biotechnology,1990,8(3):228-230
[103] Whang L M,Liu P G,Ma C C,et al.Application of biosurfactants, rhamnolipid and surfactin for enhanced biodegradation of diesel-contaminated water and soil[J].Journal of Hazardous Materials,2007,25(5):63-71
[104]傅海燕,曾光明,袁兴中,等.生物表面活性剂的分离提纯及其发展前景[J].生物学杂志,2003,20(6):1-4
[105]卢国满.产表面活性剂菌株的筛选、发酵条件优化及定量研究:[湖南大学硕士学位论文].长沙:湖南大学环境科学与工程学院,2006:8-10
[106] Cooper DG,Macdonald CR,Duff SBJ.Enhanced production of surfactin from Bacillus subtilis by continuous product removel and metal cation additions[J].Appl Environ Microbiol,1981,42(3):408-412
[107]麦有斌,尹华,叶锦韶,等.假单胞菌表面活性物主产生与性能[J].环境科学与技术,2006,29(2):12-14
[108] Velanker S K,Barneet S M,Houston C W,et al.Microbiol growth on hydrocarbons-some experimental results[J].Biotechnol Bioeng,1975,17(2): 241-251
[109] Davis D A,Lynch H C,Varley J.The production of surfactin in batch culture by Bacillus subtilis 21332 is strongly influenced by the conditions of nitrogen metabolism[J].Enzyme and Microbial Technol,1999,25(3):322-329
[110] Olness A,Clapp C G.Microbial degradation of montmorillo-nite-dextran complex[J].Soil Sci Soc AM Pro,1972,37(5):37-45
[111] Sen R,Swaminathan T.Application of response surface methodology to evaluate the optimum environmental conditions for the enhanced production of surfactin[J].Apply of Micrology Biotechnology, 997,47(8):358-363
[112] Fu H Y,Zeng G M,Huang G H,et al.Isolation of biosurfactant-producing bacteria from compost and their prospective application in composting[J].Transaction of Nonferrous Metals Societyof China,2004,14(1):131-134