|
|
|
| Isolation and identification of a high-efficient diesel degrading bacterial strain Acinetobacter sp. L7 |
| HU Chun-hui1,2, YU Hao2, ZHAO Yang-guo1, TIAN Wei-jun1, BAI Jie1 |
1. Key laboratory of Marine Environmental Science and Ecology, Ministry of Education, Ocean University of China, Qingdao 266100, China;
2. College of Life Science, Qingdao Agricultural University, Qingdao 266109, China |
|
|
|
|
Abstract A high-efficient diesel degrading bacterial strain isolated from oil-contaminated soil in Liaohe oil field was identified as Acinetobacter sp. and named as L7. The strain could efficiently grow with diesel, n-tridecane, n-hexadecane, n-heptadecane, n-octadecane and n-nonadecane as its sole carbon and energy source. The cultivation conditions of the strain L7 in diesel basic salt medium were investigated. The suitable pH value for the growth of the strain ranged from 6.0~9.0 with the optimum value of 7.0. The optimum temperature and salinity for the cultivation of the strain was 30℃ and 1%, respectively. The degradation efficiency of diesel oil was 61.5% under the optimum conditions. Based on the whole-genome DNA sequence analysis of the bacterial strain, an alkane hydroxylase gene, alkB, was identified. To study the function of the alkB gene in n-alkane utilization by the bacterial strain, the pME6032-alkB plasmid was constructed and electroporated into another bacterial strain Pseudomonas KT2440, which could not grow in diesel. The electroporated bacteria Pseudomonas KT2440 could grow in diesel basic salt medium. Through gas chromatography-mass spectrometric analysis, the degradation capacity of the bacterial strain L7 for n-alkanes and the roles of the alkB gene were determined. The results indicated that the n-alkanes were degraded by the strain L7 via the terminal oxidation and β-oxidation pathway.
|
|
Received: 05 April 2017
|
|
|
|
|
|
| [1] |
Hassanshahian M, Emtiazi G, Cappello S. Isolation and characterization of crude-oil-degrading bacteria from the Persian Gulf and the Caspian Sea[J]. Marine Pollution Bulletin, 2012, 64(1):712.
|
| [2] |
黄艺,礼晓,蔡佳良.石油污染生物修复研究进展[J]. 生态环境学报, 2009,18:361367.
|
| [3] |
Rojo F. Degradation of alkanes by bacteria[J]. Environmental Microbiology, 2009,11(10):24772490.
|
| [4] |
林莹莹,聂麦茜,王琰,等.生物膜处理高含油废水及膜表面微生物群落特性研究[J]. 中国环境科学, 2016,36(9):28002806.
|
| [5] |
张楠,初里冰,丁鹏元,等.A/O生物膜法强化处理石化废水及生物膜种群结构研究[J]. 中国环境科学, 2015,35(1):8086.
|
| [6] |
Huan L, Jing X, Rubing L, et al. Characterization of the medium-and long-chain n-alkanes degrading Pseudomonas aeruginosa strain SJTD-1and its alkane hydroxylase genes[J]. PLOS ONE, 2014,9(8):e105506.
|
| [7] |
刁硕,王红旗,许洁,等.低温耐盐芘降解菌的筛选鉴定及降解特性研究[J]. 中国环境科学, 2017,37(2):677685.
|
| [8] |
Shigeaki H, Yuki K, Akihiro H. Microbial communities in oil-contaminated seawater[J]. Current Opinion Biotechnology, 2004,15(3):205214.
|
| [9] |
Van Beilen J B, Funhoff E G. Alkane hydroxylases involved in microbial alkane degradation[J]. Applied Microbiology and Biotechnology, 2007,74(1):1321.
|
| [10] |
Diogo J, Vanessa M A, Raquel P, et al. The use of a combination of alkB primers to better characterize the distribution of alkane-degradign bacteria[J]. PLOS ONE, 2013,8(6):e66565.
|
| [11] |
Pearson W R, Lipman D J. Improved tools for biological sequence comparison[J]. Proceedings of the National Academy of Science of the United States of America, 1988,85(8):24442448.
|
| [12] |
Shan Z Y, Xu H J, Shi X Q, et al. The study of optimal conditions of electroporation in Pseudomonas aeruginosa[J]. Acta Genetica Sinica, 2004,31(3):311316.
|
| [13] |
黄延林,宁亚平,柴蓓蓓.石油高效降解菌的筛选及其降解特性[J]. 西安建筑科技大学学报(自然科学版), 2006,38(6):795803.
|
| [14] |
Wang X B, Chi C Q, Nie Y, et al. Degradation of petroleum hydrocarbons (C6-C40) and crude oil by a novel Dietzia strain[J]. Bioresource Technology, 2011,102(17):77557761.
|
| [15] |
Andreas R, Walter G, Wolfgang H. Alkane hydroxylase from Acinetobacter sp. ADP1is encoded by alkM and belongs to a new family of bacterial integral-membrane hydrocarbon hydroxylases[J]. Applied and Environment Microbiology, 1998,64(4):11751179.
|
| [16] |
Van Hamme J D, Singh A, Ward O P. Recent advances in petroleum microbiology[J]. Microbiology and Molecular Biology Reviews, 2003,67(4):503549.
|
| [17] |
Maeng J H, Sakai Y, Tani Y. Isolation and characterization of a novel oxygenase that catalyzes the first step of n-alkane oxidation in Acinetobacter spstrain M-1[J]. Journal of Bacteriology, 1996,178(13):36953700.
|
| [18] |
Mimmi T H, Alexander W, Trond E, et al. Identification of novel genes involved in long-chain n-alkane degradation by Acinetobacter sp. strain DSN 17874[J]. Applied and Environmental Microbiology, 227,73(10)33273332.
|
| [19] |
Mercedes M M, Luis Y, Fernando R. Differential expession of the components of the two alkane hydroxylases from Pseudomonas aeruginosa[J]. Journal of Bacteriology, 2003,185(10):32323237.
|
| [20] |
Cappelltti M, Fedi S, Frascari D, et al. Analyses of both the alkB gene transcriptional atart site and alkB promoter-inducing properties of Rhpdococcus sp. strain BCP1grown on n-alkanes[J]. Applied and Environment Microbiology, 2011,77(5):16191627.
|
| [21] |
Liu H, Xu J, Liang R B, et al. Characterization of the medium-and long-chain n-alkanes degrading Pseudomonas aeruginosa strain SJTD-1and its alkane hydroxylase genes[J]. PLOS ONE, 2014,9(8):e105506.
|
| [22] |
陆洪省,魏文超,王厚伟,等.高效石油烃降解菌SKD-1的分离、筛选及其降解性能[J]. 环境工程学报, 2013,7(10):41164120.
|
|
|
|
