高效石油降解菌去除石油污染土壤中的油
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摘要
随着现代生物技术的发展,微生物在环境保护方面的应用前景越来越广阔。本文就微生物在石油污染土壤的石油降解领域进行了菌种的选育、菌株的鉴定、S92菌株微生物学特性、石油降解微生物的紫外诱变、营养平衡与降解机理、以及高效降解菌群在土壤修复中的应用进行了初步研究。
本文通过一系列试验,从多种样品中广泛筛选石油降解菌,共筛选得石油降解效果较好的石油降解菌13株,分别为S43、S67、S85、S92、S104、S109、S120、S129、S136、S138、S145、S155、S167。S进一步鉴定,确认为假单胞菌属。其中石油降解效果最好的S92对模拟石油降解的降解率为87.0%。
以S92菌株为原菌株通过紫外诱变得到一株石油降解能力提高显著的菌株S-3-15。用此菌株对石油污染土样进行石油降解试验,固液质量比为40%,接种量20%,温度37℃,摇床转速150 r/min,好氧培养15 d,降解率为96.1%。因此,菌株S-3-15将作为以后的石油降解研究试验的菌源。
研究了土壤中石油烃污染物生物降解过程中营养物质供给的平衡关系及其影响降解的机理。对样品按1:45接种,加入氮,磷营养物,在37℃下恒温振荡培养。试验研究结果表明:氮、磷源同时添加效果是最好的,氮、磷元素合适的浓度分别为80 mg/L、20 mg/L.按质量计算的N、P元素的添加比例N:P为4:1。
利用菌群生物强化技术能有效提高土壤修复效率,与自然条件下的土壤修复相比,15 d内利用生物强化的降解率达到了94.2%,比自然条件下的降解率提高了38.75%。
影响上壤生物修复的因素有温度、氮磷浓度、通氧量等,通过实验得出:随着温度的升高,土壤修复效率增加,生物生长活跃,而低温条件下生物生长缓慢,即使增加通氧量也不能提高降解效果;投加氮磷营养物质能促进微生物生长,提高土壤修复效率。
石油降解微生物石油降解运行成本低,不产生二次污染,工艺简单,既有经济效益和环境效益,又具有重要的现实意义。
With the development of modern biotechnology,more and more microorganism is used in environmental protection.The screening and determinant of microbial for oil degradation,the characteristic of strain S92,the biotransformation conditions of oil degradation,and the mechanism of Strain S92 and the soil bioremediation by microorganism were investigated.
Several sorts of strains were isolated from samples by a series of experiments, 13 of them proved to be satisfactory effect on oil degradation were obtained.The best strain,named as S92 in this paper,with a higher oil degradation efficiency,was identified Pseudomons.
S92 was mutagenized to S-3-15 by ultraviolet radiation.The optimum conditions for oil degradation was as follows:ratio of solid to liquid 2:5,inoculation capacity 20%,temperature 37℃,shaking table rotating speed 150 r/min,and aerobic cultivation for 15 d.Under these conditions,an oil degradation rate of 96.1%was gained by using S-3-15.
The nutrient balance and mechanism were studied for enhancing the bioremediation of petroleum contaminated soils.The results show that the suitable condition was inoculation to sample at 1:45,And at a rotating speed of 150rpm,30℃of temperature,and 15d for cultivation,the weight ratio of N and P element in the nutrient is 4:1.
The effects of different conditions on the bioremediation were studied.The results show that.During the process of 15-day bioremediation,bioaugmentation improved the degradation rate by 94.2%compared with that in nature condition. Microorganism will be more active with the increase of temperature,so does the degradation rate;adding nutrition and oxygen content will also be contributed to microorganism growth and in the end enhance the biodegradation.
Therefore,with the advantage of low energy waste,higher efficiency, non-poisonous,harmless and non-secondary pollution,the microorganism oil degradation for polluted soil by oil is worth regarding and a further research.
本文通过一系列试验,从多种样品中广泛筛选石油降解菌,共筛选得石油降解效果较好的石油降解菌13株,分别为S43、S67、S85、S92、S104、S109、S120、S129、S136、S138、S145、S155、S167。S进一步鉴定,确认为假单胞菌属。其中石油降解效果最好的S92对模拟石油降解的降解率为87.0%。
以S92菌株为原菌株通过紫外诱变得到一株石油降解能力提高显著的菌株S-3-15。用此菌株对石油污染土样进行石油降解试验,固液质量比为40%,接种量20%,温度37℃,摇床转速150 r/min,好氧培养15 d,降解率为96.1%。因此,菌株S-3-15将作为以后的石油降解研究试验的菌源。
研究了土壤中石油烃污染物生物降解过程中营养物质供给的平衡关系及其影响降解的机理。对样品按1:45接种,加入氮,磷营养物,在37℃下恒温振荡培养。试验研究结果表明:氮、磷源同时添加效果是最好的,氮、磷元素合适的浓度分别为80 mg/L、20 mg/L.按质量计算的N、P元素的添加比例N:P为4:1。
利用菌群生物强化技术能有效提高土壤修复效率,与自然条件下的土壤修复相比,15 d内利用生物强化的降解率达到了94.2%,比自然条件下的降解率提高了38.75%。
影响上壤生物修复的因素有温度、氮磷浓度、通氧量等,通过实验得出:随着温度的升高,土壤修复效率增加,生物生长活跃,而低温条件下生物生长缓慢,即使增加通氧量也不能提高降解效果;投加氮磷营养物质能促进微生物生长,提高土壤修复效率。
石油降解微生物石油降解运行成本低,不产生二次污染,工艺简单,既有经济效益和环境效益,又具有重要的现实意义。
With the development of modern biotechnology,more and more microorganism is used in environmental protection.The screening and determinant of microbial for oil degradation,the characteristic of strain S92,the biotransformation conditions of oil degradation,and the mechanism of Strain S92 and the soil bioremediation by microorganism were investigated.
Several sorts of strains were isolated from samples by a series of experiments, 13 of them proved to be satisfactory effect on oil degradation were obtained.The best strain,named as S92 in this paper,with a higher oil degradation efficiency,was identified Pseudomons.
S92 was mutagenized to S-3-15 by ultraviolet radiation.The optimum conditions for oil degradation was as follows:ratio of solid to liquid 2:5,inoculation capacity 20%,temperature 37℃,shaking table rotating speed 150 r/min,and aerobic cultivation for 15 d.Under these conditions,an oil degradation rate of 96.1%was gained by using S-3-15.
The nutrient balance and mechanism were studied for enhancing the bioremediation of petroleum contaminated soils.The results show that the suitable condition was inoculation to sample at 1:45,And at a rotating speed of 150rpm,30℃of temperature,and 15d for cultivation,the weight ratio of N and P element in the nutrient is 4:1.
The effects of different conditions on the bioremediation were studied.The results show that.During the process of 15-day bioremediation,bioaugmentation improved the degradation rate by 94.2%compared with that in nature condition. Microorganism will be more active with the increase of temperature,so does the degradation rate;adding nutrition and oxygen content will also be contributed to microorganism growth and in the end enhance the biodegradation.
Therefore,with the advantage of low energy waste,higher efficiency, non-poisonous,harmless and non-secondary pollution,the microorganism oil degradation for polluted soil by oil is worth regarding and a further research.
引文
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[14]Aries E.Doumenq P.Artaud J.et al.Effects of petroleum hydrocarbons 013.the phospholipid fatty acid composition of a—consortium composed of marinehydrocarbon—degrading bacteria[J].Organic Geochem.,2001,32:891—903.
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[24]邓振山,任桂梅,张宝成等.陕北油田降解石油微生物的筛选及其降解能力[J].延安大学学报,2005.3(24)65-67.
[25]谢丹平,尹华,彭辉.石油降解菌株的分离及其降解特性研究[J].上海环境科学,2003,12(22)951-954.
[26]Owen Ward,Ajay Singh,J.van Hamme.Accelerated biodegradation of petroleum hydrocarbon waste[J].J of Microbio Biotechnol.2003,(30):260-270.
[27]谢丹,平尹,华彭.石油降解菌株的分离及其降解特性研究[J].上海环境科学.2003,22(12):951-954.
[28]郭书海,张海荣,张春桂.陈化石油污染物降解菌的筛选[J].农业环境科学学报.2005,24(1):161-16.
[29]赵璇,陶雪琴,卢桂宁等.萘降解菌的筛选及降解性能的初步研究[J].广东化工.2006,4(33):37-39.
[30]贺延龄,陈爱侠.环境微生物学[M].北京:中国轻工业出版社,2001,56.
[31]无锡轻工学院,华南工学院,天津轻工学院,大连轻工学院合编.微生物学[M].中国轻工业出版社,1983
[32]姚晓惠,刘秀花,梁峰.土壤中磷细菌的筛选和鉴定[J].河南农业科学,2002,(7):28-31.
[33]张爱华.巨大芽胞杆菌PS01P菌株突变株的诱变选育及其发酵条件的研究[D].南京:南京师范大学,2004:27-32.
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[2]齐永强,王红旗.微生物处理土壤石油污染的研究进展[J].上海环境科学,2002,2(3):177-195.
[3]Watanabe K,Kodama Y,Syutsubo K.Molecular characterization of bacterial populations in petroleum—contaminated groundwaterdischarged from underground crude oil storage cavities[J].Appl.Environ.Microbio1.2000,66(11):4803-4809.
[4]郑远扬编译.石油污染生化治理的进展[J].国外环境科学技术:1993.3.
[5]王保军译.石油工业油渣在土壤中的处理[J].治理技术:1998
[6]Aries E.Doumenq P.Artaud J.et al.Effects of petroleum hydrocarbons 013.the phospholipid fatty acid composition of a—consortium composed of marinehydrocarbon—degrading bacteria[J].Organic Geochem.,2001,32:891—903.
[7]王连生.有机污染化学进展[M].南京:南京大学出版社,1991.
[8]夏平,李学亚,刘斌.石油污染的生物修复[J].污染防治技术.2006,19(3):37-40
[9]陈坚.任洪强.环境生物技术运用于发展[M].北京:中国轻工业出版社,2001.
[10]沈德中.污染环境的生物修复[M].北京:化学工业出版社,2003.
[11]齐永强,王红旗.微生物处理土壤石油污染的研究进展[J].上海环境科学.2002,21(3):177-195
[12]齐永强,王红旗.微生物处理土壤石油污染的研究进展[J].上海环境科学,2002,21(3):177-195.
[13]Watanabe K,Kodama Y,Syutsubo K.Molecular characterization of bacterial populations in petroleum—contaminated groundwaterdischarged from underground crude oil storage cavities[J].App1.Environ.Microbiol.2000,66(11):4803-4809.
[14]Aries E.Doumenq P.Artaud J.et al.Effects of petroleum hydrocarbons 013.the phospholipid fatty acid composition of a—consortium composed of marinehydrocarbon—degrading bacteria[J].Organic Geochem.,2001,32:891—903.
[15]王连生.有机污染化学进展[M].南京大学出版社1991年
[16]马文漪.杨柳燕.环境微生物工程[M].南京:南京大学出版社.1998.128-131.
[17]Owen Ward,Ajay Singh,J.Van Hamme.Accelerated biodegradation of petroleum hydrocarbon waste[J].J of Microbiol Biotechnol.2003,30:260-270.
[18]李凯峰,温青,夏淑梅.石油污染土壤的生物处理技术[J].应用科技.2002,19(10):62-64.
[19]赵留辉,马娟,杨建涛.浅析石油污染土壤的处理技术[J].甘肃科技.2003,19(10):36-37.
[20]沈德中.污染物的原位生物处理一生物清消技术[J].环境科学进展,1993,11(5):56-59.
[21]秦煜民 隋智慧 魏有权.应用生物修复技术处理石油污染土壤[J].石油与天然气化工.2002,31(6):333-335.
[22]王辉,赵春燕,李宝明,孙军德.石油污染土壤中细菌的分离筛选[J].土壤通报,2005,36(2)237-239.
[23]周长征.李秀云.宋延博.三维荧光法在石油污染鉴别中的应用[J].光谱学与光谱分析.1998,18(1):500-502
[24]邓振山,任桂梅,张宝成等.陕北油田降解石油微生物的筛选及其降解能力[J].延安大学学报,2005.3(24)65-67.
[25]谢丹平,尹华,彭辉.石油降解菌株的分离及其降解特性研究[J].上海环境科学,2003,12(22)951-954.
[26]Owen Ward,Ajay Singh,J.van Hamme.Accelerated biodegradation of petroleum hydrocarbon waste[J].J of Microbio Biotechnol.2003,(30):260-270.
[27]谢丹,平尹,华彭.石油降解菌株的分离及其降解特性研究[J].上海环境科学.2003,22(12):951-954.
[28]郭书海,张海荣,张春桂.陈化石油污染物降解菌的筛选[J].农业环境科学学报.2005,24(1):161-16.
[29]赵璇,陶雪琴,卢桂宁等.萘降解菌的筛选及降解性能的初步研究[J].广东化工.2006,4(33):37-39.
[30]贺延龄,陈爱侠.环境微生物学[M].北京:中国轻工业出版社,2001,56.
[31]无锡轻工学院,华南工学院,天津轻工学院,大连轻工学院合编.微生物学[M].中国轻工业出版社,1983
[32]姚晓惠,刘秀花,梁峰.土壤中磷细菌的筛选和鉴定[J].河南农业科学,2002,(7):28-31.
[33]张爱华.巨大芽胞杆菌PS01P菌株突变株的诱变选育及其发酵条件的研究[D].南京:南京师范大学,2004:27-32.
[34]Atlas R M.Microbial degradation of petroleum hydrocarbons:an environmental perspective[J].Microb.,1981,45:180-209.
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