产表面活性剂石油降解菌的筛选及其降解率的测定
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摘要
为了筛选出具有高效降解石油能力的菌株,从大庆油田石油污染土壤中分离出石油降解菌株,通过血平板筛选对其进行产表面活性剂能力的初步测定,并利用紫外法和称重法来测定其降解石油的能力。其中低温石油降解菌株DDX82的溶血圈直径为19.47 mm,紫外法测定其降解率为60.69%,称重法测定其降解率为47.92%,较其他石油降解菌株产表面活性剂的能力更强,降解石油的效果更明显,经革兰氏染色、形态学观察、生理生化及16SrDNA序列分析等,确定菌属于假单胞菌属菌株。实验表明,该菌株的最佳降解温度为20℃,在此条件下,DDX82第8天时降解率最高达到64.98%。
In order to screen strains which were good at petroleum degradation,an oil-degrading bacteria was isolated from petroleum-contaminated soil of Daqing Oil Field in Heilongjiang province,and its ability of production surfactant was preliminarily determined by blood plate screening,and its petroleum degradation ability was tested using ultraviolet and weighing method.The results revealed that the DDX82's hemolytic circle diameter was 19.47 mm,the degradation rate by UV method was 60.69%,and by weighing method was 47.92%.Compared to other oil-degrading strains,the surfactant production ability of DDX82 was much stronger and was more efficient on oil degradation.After the Gram stain,morphological observation,physiological and biochemical and 16 S rDNA sequence analysis,the strain was identified as pseudomonas monteilii.The results showed that the optimal degradation conditions were as followings:the temperature was 20 ℃,the degradation rate reached 64.98% at the 8 th day.
In order to screen strains which were good at petroleum degradation,an oil-degrading bacteria was isolated from petroleum-contaminated soil of Daqing Oil Field in Heilongjiang province,and its ability of production surfactant was preliminarily determined by blood plate screening,and its petroleum degradation ability was tested using ultraviolet and weighing method.The results revealed that the DDX82's hemolytic circle diameter was 19.47 mm,the degradation rate by UV method was 60.69%,and by weighing method was 47.92%.Compared to other oil-degrading strains,the surfactant production ability of DDX82 was much stronger and was more efficient on oil degradation.After the Gram stain,morphological observation,physiological and biochemical and 16 S rDNA sequence analysis,the strain was identified as pseudomonas monteilii.The results showed that the optimal degradation conditions were as followings:the temperature was 20 ℃,the degradation rate reached 64.98% at the 8 th day.
引文
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[24]郝婧,高磊,吴春旭,等.石油降解菌Pseudomonas stutzeri TH-31的分离与降解条件优化[J].环境工程学报,2015,9(4):1771-1777.
[25]任兵,张金艳,李绍峰.2株西玛津降解菌的降解特性研究[J].黑龙江八一农垦大学学报,2016,28(3):99-104.
[26]张月梅,祖国仁,那广水,等.北极耐冷石油降解菌的筛选、鉴定及其碳源利用广谱性[J].海洋环境科学,2010,29(2):217-220.
[27]刘秋,张耀尹,曹雪洁,等.海洋石油降解微生物及其降解机理[J].微生物学杂志,2016(36):1-6.
[28]Atlas RM.Petroleum biodegradation and oil spill bioremediation[J].Marine Pollution Bulletin,1995,31(4):178-182.
[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,2002,21(3):177-180.
[3]祝威.石油污染土壤和油泥生物处理技术[M].北京:中国石化出版社,2010.
[4]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.
[5]Meudec A,Poupart N,Dussauze J,et al.Relationship between heavy fuel oil phytotoxicity and plycyc-lic aromatic hydrocarbon contamination in Salicornia fragilis[J].Science of the Total Environment,2007,381(1-3):146-156.
[6]郭军权,王文龙,刘恩斌,等.石油污染土壤生物治理技术研究进展[J].环境管理,2013(3):73-75.
[7]李春荣,王文科,曹玉清,等.石油污染物的微生物降解研[J].生态环境,2008,17(1):113-116.
[8]马飞,潘科,陶文,等.一株产表面活性剂的高效烃降解菌的筛选[J].广东石油化工学院学报,2012,22(1):22-25.
[9]陆昕.产表面活性剂石油降解菌的选育及对陕北石油污染土壤生物修复的实验研究[D].西安:西北大学,2010.
[10]王靖,陈云.生物表面活性剂及其评价方法[J].化学研究与应用,2009,21(2):137-140.
[11]刘桂萍,刘巍巍,刘文杰,等.生物表面活性剂产生菌的筛选及培养条件优化[J].环境保护科学,2011,37(6):12-15.
[12]赵辉,闺华晓,杨腾,等.高效生物表面活性剂产生菌筛选及其性质研究[J].生物技术通报,2010,20(4):76-78.
[13]张娜,郑青松,刘玲,等.产表面活性剂石油降解菌的筛选及发酵条件优化[J].南京农业大学学报,2011,34(4):133-137.
[14]沈萍,范秀容,李广武.微生物学实验[M].北京:高等教育出版社,2004.
[15]鲁雅梅,邹东雷,赵永胜,等.烷烃降解菌SY16的筛选,鉴定及降解能力测定[J].生态环境,2007,16(5):1394-1398.
[16]丁立孝,何国庆,刘晔,等.脂肽生物表面活性剂产生菌的筛选[J].农业生物技术学,2004,12(3):330-333.
[17]包木太,田艳敏,陈庆国.海藻酸钠包埋固定化微生物处理含油废水研究[J].环境科学与技术,2012,35(2):167-172.
[18]奚旦立,孙裕生,刘秀英,等.环境监测(修订版)[M].北京:高等教育出版社,1995.
[19]国家环保局《水和废水监测分析方法》编写组.水和废水监测分析方法[M].4版.北京:中国环境科学出版社,2002.
[20]Holt JG,Krieg NR,Sneath PHA,et al.Bergey's Manual of detective bacteriology[M].Maryland:Williams&Wilkins Baltimore,1994.
[21]夏铁骑,常慧萍,张建清.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].农业灾害研究,2013(6):49-51.
[22]Abalos A,Vinas M,Sabate J,et al.Enhanced biodegradation of Casablanca crude oil by a microbal consortium in presence of a rhamnolipid produced by Pseudomonas aerufinosa AT10[J].Biodegradation,2004,15(4):260.
[23]慕庆峰,识音.石油降解细菌的筛选及培养条件的初步研究[J].黑龙江八一农垦大学学报,2008,28(3):19-22.
[24]郝婧,高磊,吴春旭,等.石油降解菌Pseudomonas stutzeri TH-31的分离与降解条件优化[J].环境工程学报,2015,9(4):1771-1777.
[25]任兵,张金艳,李绍峰.2株西玛津降解菌的降解特性研究[J].黑龙江八一农垦大学学报,2016,28(3):99-104.
[26]张月梅,祖国仁,那广水,等.北极耐冷石油降解菌的筛选、鉴定及其碳源利用广谱性[J].海洋环境科学,2010,29(2):217-220.
[27]刘秋,张耀尹,曹雪洁,等.海洋石油降解微生物及其降解机理[J].微生物学杂志,2016(36):1-6.
[28]Atlas RM.Petroleum biodegradation and oil spill bioremediation[J].Marine Pollution Bulletin,1995,31(4):178-182.