一株耐低温原油降解菌的分离鉴定及降解特性

doi:10.3969/j.issn.1004-1524.2016.10.21

浙江农业学报 ›› 2016, Vol. 28 ›› Issue (10): 1781-1789.DOI: 10.3969/j.issn.1004-1524.2016.10.21

一株耐低温原油降解菌的分离鉴定及降解特性

杨蕊琪¹, 薛林贵^{1, *}, 常思静², 章高森², 陈拓², 刘光琇²

1.兰州交通大学化学与生物工程学院,甘肃省极端环境微生物资源与工程重点实验室,甘肃兰州 730070;
2.甘肃省极端环境微生物资源与工程重点实验室,中国科学院寒区旱区环境与工程研究所沙漠与沙漠化重点实验室,甘肃兰州 730000

收稿日期:2015-12-23 出版日期:2016-10-15 发布日期:2016-11-20
通讯作者: 薛林贵,E-mail:xuelg@mail.lzjtu.cn
作者简介:杨蕊琪(1989—),女,甘肃平凉人,硕士研究生,从事环境微生物学方面的研究。E-mail:514503456@qq.com
基金资助:
国家国际科技合作专项项目(2014DFA30330); 国家自然科学基金项目(31260090,31260135)

Studies on isolation, identification of cold-resistant petroleum-degrading strain and its degradation capability

YANG Rui-qi¹, XUE Lin-gui^{1, *}, CHANG Si-jing², ZHANG Gao-sen², CHEN Tuo², LIU Guang-xiu²

1. College of Chemical and Biology Engineering, Lanzhou Jiaotong University, Key Laboratory of Extreme Environmental Microbial Resources and Engineering of Gansu Province, Lanzhou 730070, China;
2. Key Laboratory of Extreme Environmental Microbial Resources and Engineering of Gansu Province, Key laboratory of Desert and Desertification, Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China

Received:2015-12-23 Online:2016-10-15 Published:2016-11-20

摘要/Abstract

摘要： 从青藏高原祁连山地区的土壤中分离筛选出一株高效耐低温原油降解菌YF28-1(8),其降解率为(57.90±3.86)%,经形态观察、生理生化及16S rRNA同源性分析进行初步鉴定,确定该菌株为红平红球菌(Rhodococcus erythropolis)。通过气相色谱—质谱联用法(GC-MS)、紫外分光光度法和重量法对原油降解菌降解性能进行研究。结果表明,降解菌YF28-1(8)对原油中的烷烃组分均有不同程度的降解,但对直链烷烃具有较强的生物降解作用。该菌株的最适降解条件为:初始pH 7.5、温度30 ℃、接种量800 μL,培养8 d后生长量及降解率达到最大值,吐温80的添加只略微提高了菌株降解率。

关键词: 低温, 原油降解菌, GC-MS, 降解特性

Abstract: We obtained a low-temperature resistant crude oil degrading strain known as YF28-1(8) from the soil samples in Qinghai-Tibet Plateau, Qilian mountain, whose degradation rate was (57.90±3.86)%. Besides, after a preliminary identification on the basis of investigation on its biochemical and biological characteristics and 16S rRNA homology analysis, we defined this strain as Rhodococcus erythropolis. In addition, we drew a conclusion about degradation capability by means of GC-MS, UV-spectrophotometry, gravimetry that YF28-1(8) was able to biodegrade all alkane constituents in different degrees and especially effective to linear saturated alkane. The initial optimum conditions of this strain included: pH value at 7.5, temperature at 30 ℃ and inoculation size of 800 μL and its quantity and degradation rate could reach maximum in only 8 days, meanwhile, surfactant named Tween 80 could only slightly improve its degradation rate.

Key words: low-temperature, crude oil degrading bacterial, GC-MS, degradation capability

中图分类号:

S476.1

杨蕊琪, 薛林贵, 常思静, 章高森, 陈拓, 刘光琇. 一株耐低温原油降解菌的分离鉴定及降解特性[J]. 浙江农业学报, 2016, 28(10): 1781-1789.

YANG Rui-qi, XUE Lin-gui, CHANG Si-jing, ZHANG Gao-sen, CHEN Tuo, LIU Guang-xiu. Studies on isolation, identification of cold-resistant petroleum-degrading strain and its degradation capability[J]. , 2016, 28(10): 1781-1789.

参考文献

[1] HEAD I M, JONES D M, RÖLING W F M. Marine microorganisms make a meal of oil[J]. Nature Reviews Microbiology , 2006, 4(3): 173-182.
[2] 金会军,喻文兵,陈友昌,等. 多年冻土区输油管道工程中的(差异性)融沉和冻胀问题[J]. 冰川冻土, 2005, 27(4): 454-464.
JIN H J, YU W B, CHEN Y C, et al. (Differential)frost heave and thaw settlement in the engineering design and construction of oil pipelines in permafrost regions: A review[J]. Journal of Glaciology and Geocryology , 2005, 27(3): 454-464. (in Chinese with English abstract)
[3] 李潞滨,刘振静,杨凯,等. 青藏铁路沿线唐古拉山口土壤微生物的ARDRA分析[J]. 生态学报, 2008, 28(11): 5482-5487.
LI L B, LIU Z J, YANG K, et al. Soil microbial ARDRA analysis of Tanggula Mountain pass along the Qinghai-Tibet railway[J]. Acta Ecologica Sinica , 2008, 28(11): 5482-5487. (in Chinese with English abstract)
[4] 徐冯楠,冯贵颖,马雯,等. 高效石油降解菌的筛选及其降解性能研究[J]. 生物技术通报, 2010, (7): 221-226.
XU F N, FENG G Y, MA W, et al. Study on the screening and degradation characteristics of highly efficient petroleum degrading bacteria[J]. Biotechnology Bulletin , 2010, (7): 221-226. (in Chinese with English abstract)
[5] ESMAEIL A L S, DROBIOVA H, OBUEKWE C. Predominant culturable crude oil-degrading bacteria in the coast of Kuwait[J]. International Biodeterioration & Biodegradation , 2008, 63(4): 400-406.
[6] JURELEVICIUS D, ALVAREZ V M, PEIXOTO R, et al. The use of a combination of alkB primers to better characterize the distribution of alkane-degrading bacteria[J]. PloS One , 2013, 8(6): e66565.
[7] GUIBERT L M, LOVISO C L, MARCOS M S, et al. Alkane biodegradation genes from chronically polluted subantarctic coastal sediments and their shifts in response to oil exposure[J]. Microbial Ecology , 2012, 64(3): 605-616.
[8] JURELEVICIUS D, COTTA S R, PEIXOTO R, et al. Distribution of alkane-degrading bacterial communities in soils from King George Island, Maritime Antarctic[J]. European Journal of Soil Biology , 2012, 51: 37-44.
[9] 王红旗,郝旭光,孙寓姣,等. 土壤中耐低温石油降解菌的优选、鉴定及降解性能分析[J]. 北京师范大学学报(自然科学版), 2009, 45(5/6): 509-514.
WANG H Q, HAO X G, SUN Y J, et al. Selection, identification and degradation characteristics of cold-resistant petroleum-degrading strains in soil[J]. Journal of Beijing Normal University ( Natural Science ), 2009, 45(5/6): 509-514. (in Chinese with English abstract)
[10] 金铭,车宗玺,敬文茂,等. 祁连山水源涵养林区降水及温度时空变化研究[J]. 中国沙漠, 2012, 32(4): 1071-1076.
JIN M, CHE Z X, JING W M, et al. Spatiotemporal changes of precipitation and temperature at water resources conservation mountains, China[J]. Journal of Desert Research , 2012, 32(4): 1071-1076. (in Chinese with English abstract)
[11] 贾燕,尹华,彭辉,等. 石油降解菌株的筛选、初步鉴定及其特性[J]. 暨南大学学报(自然科学版), 2007, 28(3): 296-301.
JIA Y, YIN H, PENG H, et al. Isolation, Identification and characteristics of crude oil-degrading microorganism[J]. Journal of Jinan University ( Natural Science ), 2007, 28(3): 296-301. (in Chinese with English abstract)
[12] WANG W P, WANG L P, SHAO Z Z. Diversity and abundance of oil-degrading bacteria and alkane hydroxylase (alkB) genes in the subtropical seawater of Xiamen Island[J]. Microbial Ecology , 2010, 60(2): 429-439.
[13] WANG B J, LAI Q L, CUI Z S, et al. A pyrene-degrading consortium from deep-sea sediment of the West Pacific and its key member Cycloclasticus sp. P1[J]. Environmental Microbiology , 2008, 10(8): 1948-1963.
[14] DUNBAR J, TAKALA S, BARNS S M, et al. Levels of bacterial community diversity in four arid soils compared by cultivation and 16S rRNA gene cloning[J]. Applied and environmental microbiology , 1999, 65(4): 1662-1669.
[15] LANE D J. 16S/23S rRNA sequencing[C]//STACKEBRANDT E,GOODFELLOW M. Nucleic acid techniques in bacterial systematics. Chichester: Wiley, 1991: 125-175.
[16] SUN J, XU L, TANG Y, et al. Simultaneous degradation of phenol and n-hexadecane by Acinetobacter strains[J]. Bioresource Technology , 2012, 123: 664-668.
[17] 王博. 土壤石油降解菌的筛选与微生物生态修复研究[D]. 南京: 南京农业大学, 2012.
WANG B. Studies on isolation of oil-degrading bacteria and microbial remediation of oil contaminated soil[D]. Nanjing: Nanjing Agricultural University, 2012. (in Chinese with English abstract)
[18] WANG X, CHI C, NIE Y, et al. Degradation of petroleum hydrocarbons (C6-C40) and crude oil by a novel Dietzia strain[J]. Bioresource Technology , 2011, 102(17): 7755-7761.
[19] 陆健,黄潇,伍贤明,等. 高效烷烃降解菌xcz的分离鉴定及降解特性[J]. 土壤, 2008, 40(3): 460-464.
LU J, HUANG X, WU X M, et al. Isolation, identification and characterization of a high-efficiency alkane-degrading bacterium xcz[J]. Soils , 2008, 40(3): 460-464. (in Chinese with English abstract)
[20] 张鲁进,杨谦,陈中祥,等. 两株石油降解菌的降解性能研究[J]. 南京理工大学学报(自然科学版), 2010, 34(6): 849-854.
ZHANG L J, YANG Q, CHEN Z X, et al. Degreding characteristics of two petrolecm-degrading strains[J]. Journal of Nanjing University of Science and Technology ( Natural Science ), 2010, 34(6): 849-854. (in Chinese with English abstract)
[21] 齐永强,王红旗,刘敬奇. 土壤中石油污染物微生物降解过程中各石油烃组分的演变规律[J]. 环境科学学报, 2003, 23(6): 834-836.
QI Y Q, WANG H Q, LIU J Q. Component changes of hydrocarbon during the precess of bioremediation of hydrocarbon-contaminated soil[J]. Acta Scientiae Circumstantiae , 2003, 23(6): 834-836. (in Chinese with English abstract)
[22] 徐恒刚,姚秀清,李倩,等. 两种高效原油降解菌降解率测定方法的对比研究[J]. 化学与生物工程, 2006, 23(9): 43-44.
XU H G, YAO X Q, LI Q, et al. The contrast study on two methods for degradation rate of crude oil by high efficient degradation bacteria[J]. Chemistry & Bioengineering , 2006, 23(9): 43-44. (in Chinese with English abstract)
[23] 何丽媛,党志,唐霞,等. 混合菌对原油的降解及其降解性能的研究[J]. 环境科学学报, 2010, 30(6): 1220-1227.
HE L Y, DANG Z, TANG X, et al. Biodegradation characteristics of crude oil by mixed bacterial strains[J]. Acta Scientiae Circumstantiae , 2010, 30(6): 1220-1227. (in Chinese with English abstract)
[24] MARGESIN R. SCHINNER F. Effect of temperature on oil degradation by a psychrotrophic yeast in liquid culture and in oil[J]. FEMS Microbiology Ecology , 1997, 24(3): 243-249.
[25] 谢鲲鹏,周集体,曲媛媛,等. 一株耐盐原油降解菌的分离鉴定及其降解特性研究[J]. 海洋环境科学, 2009, 28(6): 680-683.
XIE K P, ZHOU J T, QU Y Y, et al. Studies on isolation, identification of halophil crude oil degrading bacteria and their degradation capability[J]. Marine Environmental Science , 2009, 28(6): 680-683. (in Chinese with English abstract)
[26] 白洁,崔爱玲,吕艳华. 石油降解菌对石油烃的降解能力及影响因素研究[J]. 海洋湖沼通报, 2007(3): 41-48.
BAI J, CUI A L, LV Y H. The degradation capability of crude oil degrading strains and the impacting factors[J]. Transactions of Oceanology and Limnology , 2007(3): 41-48. (in Chinese with English abstract)
[27] 马爱青,陈连喜,包木太. 表面活性剂对原油生物降解的强化作用[J]. 油田化学, 2011, 28(2): 224-228.
MA A Q, CHEN L X, BAO M T. Reinforcement effect of surfactants on petroleum biodegradation[J]. Oilfield Chemistry , 2011, 28(2): 224-228. (in Chinese with English abstract)
[28] 张丽芳,肖红,魏德洲. 表面活性剂对土壤石油污染物微生物降解的影响[J]. 辽宁化工, 2002, 32(12): 509-513.
ZHANG L F, XIAO H, WEI D Z. Effect of surfactants on biodegradation of petroleum-contaminated soil[J]. Liaoning Chemical Industry , 2002, 32(12): 509-513. (in Chinese with English abstract)
[29] 王冬梅,陈丽华,周立辉,等. 鼠李糖脂对微生物菌剂降解石油的影响[J]. 环境工程学报, 2013, 7(10): 4121-4126.
WANG D M, CHEN L H, ZHOU L H, et al. Effects of rhamnolipid on petroleum degradation of compound microbial inoculant[J]. Chinese Journal of Environmental Engineering , 2013, 7(10): 4121-4126. (in Chinese with English abstract)
[30] ARONSTEIN B N, CALVILLO Y M, ALEXANDER M. Effect of surfactants at low concentrations on the desorption and biodegradation of sorbed aromatic compounds in soil[J]. Environmental Science & Technology , 1991, 25(10): 1728-1731.
[31] WI O ' KOMIRSKI B, SUDNIK-WÓJCIKOWSKA B, GALERA H, et al. Railway transportation as a serious source of organic and inorganic pollution[J]. Water , Air , & Soil Pollution , 2011, 218(1/4): 333-345.

一株耐低温原油降解菌的分离鉴定及降解特性

Studies on isolation, identification of cold-resistant petroleum-degrading strain and its degradation capability

RichHTML

PDF (PC)

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	苟秉调, 段盼盼, 杨楠, 赵淑芳, 王永富, 张高原, 魏兵强. 低温弱光胁迫下辣椒苗期光合相关指标的杂种优势[J]. 浙江农业学报, 2021, 33(3): 429-436.
[2]	李甜竹, 孙玉文, 汪季涛, 甘德芳, 胡克玲. 不同贮藏温度对糙皮侧耳采后品质及木质素积累的影响[J]. 浙江农业学报, 2021, 33(2): 248-258.
[3]	许双燕, 张涛, 张成, 林辉, 水贤磊, 郑华宝. 一株红霉素降解菌的筛选、鉴定与降解特性[J]. 浙江农业学报, 2021, 33(1): 131-141.
[4]	高文静, 肖丽娇, 王顺民, 韩秋霞. 降解柴油嗜盐菌的筛选、鉴定及其降解特性[J]. 浙江农业学报, 2020, 32(7): 1241-1252.
[5]	张淑文, 梁森苗, 朱婷婷, 任海英, 郑锡良, 戚行江. 不同杨梅品种的耐低温能力比较[J]. 浙江农业学报, 2020, 32(10): 1772-1779.
[6]	梁浩花, 陶红, 王亚娟, 李娇玲. 一株邻苯二甲酸二丁酯和邻苯二甲酸二(2-乙基己基)酯降解菌的筛选鉴定与降解特性[J]. 浙江农业学报, 2019, 31(7): 1145-1153.
[7]	时羽杰, 李兴龙, 唐媛, 余海萍, 邬晓勇. 基于GC-MS分析两地白色藜麦种子的代谢差异[J]. 浙江农业学报, 2019, 31(6): 869-877.
[8]	李国丽, 曾小英, 翟立翔, 冷艳, 刘梦圆, 李师翁, 陈拓. 一株石油降解菌Lysinibacillus fusiformis 23-1的筛选鉴定及原油降解特性[J]. 浙江农业学报, 2018, 30(7): 1229-1236.
[9]	滕尧, 李安定, 郝自远, 张洪亮, 张丽敏, 蔡国俊. 西番莲解剖结构特征及低温胁迫下叶片结构与抗寒性的关系[J]. 浙江农业学报, 2018, 30(11): 1849-1858.
[10]	洪森荣, 宁本松, 叶思雨, 刘燕, 张铭心, 占学林. 黄独胚性愈伤组织冻后黑暗培养及其再生植株半薄切片观察[J]. 浙江农业学报, 2018, 30(1): 65-70.
[11]	肖文斐, 倪深, 裘劼人, 王淑珍, 忻雅, 阮松林. 水稻冷激蛋白基因OsCSP2启动子的克隆与分析[J]. 浙江农业学报, 2017, 29(6): 857-863.
[12]	洪森荣, 吴夏俊鹏, 徐文慧, 占学林, 谢妮妮, 蒋妍, 汪金华, 凌飞, 吴丽霞, 万琳. 低温离体保存黄独微型块茎转录组、蛋白质组和代谢组的关联分析[J]. 浙江农业学报, 2017, 29(11): 1827-1834.
[13]	尹明华, 周宇瑶, 杨星鹏, 徐玉琴, 刘郑英, 舒荣建, 魏志敏, 夏华炎, 洪森荣. 上饶早梨主栽品种病毒种类分析及其茎尖脱毒技术效率比较[J]. 浙江农业学报, 2017, 29(1): 89-100.
[14]	高青海，王亚坤，郭远远.. 薄皮甜瓜种质资源苗期耐低温弱光鉴定及形态指标选择[J]. 浙江农业学报, 2016, 28(8): 1360-.
[15]	蒋景龙，沈季雪，徐卫平，田雲，李丽. 外源H2O2对低温胁迫下大红柑生长及叶片生理指标的影响[J]. 浙江农业学报, 2016, 28(7): 1164-.