Diversity of PAH dioxygenase genes of activated sludge from coking wastewater
MENG Xiao-jun1,2, LI Hai-bo2, SHENG Yu-xing2, CAO Hong-bin2
1. School of Tourism and Environment, Ankang University, Ankang 725000, China;
2. Key Laboratory of Green Process and Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
In order to analyze the biodiversity of PAH dioxygenase in activated sludge from a full-scale coking wastewater, primers were used for dioxygenase gene cloning expression and diversity analysis by 16Sr DNA-PCR-DGGE method using total DNA of aerobic activated sludge as the template. The results showed that significant amplification product was found using primers RHD-GN-610F and RHD-GN-916R, and the product size was 300bp. PCR product was conducted by DGGE analysis and eight separate bands were found, the abundance was 2.99%, 8.16%, 20.75%, 28.50%, 8.62%, 7.26%, 10.62% and 13.10%, respectively. Obvious amplification products further appeared after strips by gel extraction and PCR amplification, and 4sequences were tested successfully by TA cloning, the length of the sequences was 305bp, 298bp, 334bp and 294bp, respectively, indicating the presence of different PAH RHD enzymes in coking activated sludge. These results provide a theoretical basis for the risk assessment and potential biodegradation of PAHs.
Qiao M, Qi W X, Liu H J, et al. Occurrence, behavior and removal of typical substituted and parent polycyclic aromatic hydrocarbons in a biological wastewater treatment plant[J]. Water Research., 2014,52:11-19.
[2]
Yao M, Zhang X W, Lei L C. Polycyclic aromatic hydrocarbons in the centralized wastewater treatment plant of a chemical industry zone:removal, mass balance and source analysis[J]. Science China Chemistry, 2012,55(3):416-425.
[3]
Tian W J, Bai J, Liu K K, et al. Occurrence and removal of polycyclic aromatic hydrocarbons in the wastewater treatment process[J]. Ecotoxicology and Environmental Safety, 2012,82:1-7.
[4]
Zhang Y X, Tao S. Global atmospheric emission inventory of polycyclic aromatic hydrocarbons for 2004[J]. Atmospheric Environment, 2009,43:812-819.
[5]
Mu L, Peng L, Cao J J, et al. Emissions of polycyclic aromatic hydrocarbons from coking industries in China[J]. Particuology, 2013,11:86-93.
[6]
Ma W, Li Y, Qi H, et al. Seasonal variations of sources of polycyclic aromatic hydrocarbons (PAHs) to a northeastern urban city, China[J]. Chemosphere, 2010,79(4):441-447.
[7]
Pavlovich L B, Zhuravleva N V, Bal'tser D V. Polycyclic aromatic hydrocarbons in coke plant wastewater[J].Coke and Chemistry, 2010,53(10):390-395.
[8]
Zhang W H, Wei C H, Chai X S, et al. The behaviors and fate of polycyclic aromatic hydrocarbons (PAHs) in a coking wastewater treatment plant[J]. Chemosphere, 2012,88(2):174-182.
[9]
Vaessen H A M G, Jekel A A, Wilbers A A M M. Dietary intake of polycyclic aromatic hydrocarbons[J]. Toxicologiacl and Environmental Chemistry, 1988,16:281-287.
Deng L J, Ren Y, Wei C H. Pyrene degradation by Pseudomonas sp. and Burkholderia sp. enriched from coking wastewater sludge[J]. Journal of Environmental Science and Health, part A, 2012, 47:1984-1991.
[12]
Klankeo P, Nopcharoenkul W, Pinyakong O, et al. Two novel pyrene-degrading Diaphorobacter sp. and Pseudoxanthomonas sp. isolated from soil[J]. Journal of Bioscience and Bioengineering, 2009,108(6):488-495.
[13]
Balachandran C, Duraipandiyan V, Balakrishna K,et al.Petroleum and polycyclic aromatic hydrocarbons (PAHs) degradation and naphthalene metabolism in Streptomyces sp. (ERI-CPDA-1) isolated from oil contaminated soil[J]. Bioresource Technology, 2012,112:83-89.
[14]
Zhang J, Zhang X, Liu J, et al. Isolation of a thermophilic bacterium, Geobacillus sp. SH-1, capable of degrading aliphatic hydrocarbons and naphthalene simultaneously and identification of its naphthalene degrading pathway[J]. Bioresource Technology, 2012,124:83-89.
[15]
Lin M, Hu X K, Chen W W, et al. Biodegradation of phenanthrene by Pseudomonas sp. BZ-3, isolated, from crude oil contaminated soil[J]. International Biodeterioration & Biodegradation, 2014,94:176-181.
[16]
Tao X Q, Lu G N, Dang Z, et al. A phenanthrene-degrading strain Sphingomonas sp. GY2B isolated from contaminated soils[J]. Process Biochemistry, 2007,42:401-408.
[17]
Wongwongsee W, Chareanpat P, Pinyakong O. Abilities and genes for PAH biodegradation of bacteria isolated from mangrove sediments from the central of Thailand[J]. Marine Pollution bulletin, 2013,74:95-104.
[18]
Peng R H, Xiong A S, Xue Y, et al. Microbial biodegradation of polyaromatic hydrocarbons[J]. FEMS Microbiology Reviews, 2008,32:927-955.
[19]
Zhou H W, Guo C L, Wong, Y S, et al. Genetic diversity of dioxygenase genes in polycyclic aromatic hydrocarbon-degrading bacteria isolated from mangrove sediments[J]. FEMS Microbiology Letters, 2006,262:148-157.
[20]
Gomes N C M, Borges L R, Paranhos R, et al. Diversity of nod genes in mangrove sediments exposed to different sources of polycyclic aromatic hydrocarbon pollution[J]. Applied Environmental Microbiology, 2007,73:7392-7399.
[21]
Cébron A, Norini M P, Beguiristain T, et al. Real-Time PCR quantification of PAH-ring hydroxylating dioxygenase (PAH- RHDα) genes from Gram positive and Gram negative bacteria in soil and sediment samples[J]. Journal of Microbiological Methods, 2008,73:148-159.
[22]
Ding G C, Heuer H, Zuhlke S, et al. Soil type-dependent responses to phenanthrene as revealed by determining the diversity and abundance of polycyclic aromatic hydrocarbon ring-hydroxylating dioxygenase genes by using a novel PCR detection system[J]. Applied and Environmental Microbiology, 2010,76(14):4765-4771.
Deng L J, Ren Y, Wei C H. Pyrene degradation by Pseudomonas sp. and Burkholderia sp. enriched from coking wastewater sludge[J]. Journal of Environmental Science and Health, part A, 2012,47:1984-1991.
[27]
Ma Q, Qu Y Y, Shen W L, et al. Bacterial community compositions of coking wastewater treatment plants in steel industry revealed by Illumina high-throughput sequencing[J]. Bioresource Technology, 2015,179:436-443.
[28]
Klankeo P, Nopcharoenkul W, Pinyakong O. Two novel pyrene-degrading Diaphorobacter sp. and Pseudoxanthomonas sp. isolated from soil[J]. Journal of Bioscience and Bioengineering, 2009,108:488-495.
