反应器进水管生物膜中喹啉降解菌的分离鉴定及降解特性
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摘要
【背景】喹啉是一类高毒、致癌且难降解的含氮杂环化合物,本实验室建立了一个长期高效运行的反硝化喹啉降解生物反应器。【目的】从反应器进水管富集的生物膜中筛选有氧条件下降解喹啉的菌株。【方法】通过以喹啉为唯一碳源的培养基来富集、分离、纯化菌株;利用16S rRNA基因的序列分析鉴定分离株的系统发育地位;比较不同pH及温度条件下菌株的喹啉降解特性。【结果】经鉴定,4株分离物Q1、Q3、Q7和Q8分别属于Sphingobium、Massilia、Rhodococcus和Dyadobacter属。降解实验表明,以上菌株均能在48 h内实现50 mg/L喹啉的完全去除,但各自表现出不同的降解特性,其中Q1、Q3和Q8在降解过程中都检测到了喹啉降解产物2-羟基喹啉的积累。降解喹啉的Sphingobium、Massilia和Dyadobacter属菌株尚未见报道。【结论】从喹啉降解生物反应器的进水管内分离的4株喹啉降解菌可为设计处理含喹啉工业废水的反应器提供新菌种资源,对于完善喹啉生物降解机理研究具有实际意义。
[Background] Quinoline is a nitrogen heterocyclic compound with high toxicity,carcinogenicity but low degradability. A lab-scale denitrifying quinoline-degrading bioreactor was developed and in operation for several years. [Objective] To isolate the potential quinoline-degrading bacteria from the influent pipeline of the bioreactor. [Methods] The media with quinoline as sole carbon source were adopted to enrich, screen and purify degrading strains. Phylogenetic analysis was performed through 16 S rRNA gene sequencing. The characteristics of quinoline degradation by different strains were investigated under different pH and temperature conditions. [Results] Based on the phylogenetic analysis,four isolates Q1, Q3, Q7 and Q8 were identified as the genera of Sphingobium, Massilia, Rhodococcus and Dyadobacter, respectively. We demonstrated that 50 mg/L quinoline could be removed within 48 h by all of the strains with varied degradation characteristics. The accumulation of 2-hydroxyquinoline was detected during the cultivation of strain Q1, Q3 and Q8. Notably, the genera of Sphingobium, Massilia and Dyadobacter have not been reported yet on quinoline degradation. [Conclusion] The four quinoline-degrading bacteria isolated from the influent pipeline of bioreactor can provide novel strain resources for decontaminating industrial wastewater containing quinoline, which may contribute to further understanding of the mechanism of quinoline biodegradation.
[Background] Quinoline is a nitrogen heterocyclic compound with high toxicity,carcinogenicity but low degradability. A lab-scale denitrifying quinoline-degrading bioreactor was developed and in operation for several years. [Objective] To isolate the potential quinoline-degrading bacteria from the influent pipeline of the bioreactor. [Methods] The media with quinoline as sole carbon source were adopted to enrich, screen and purify degrading strains. Phylogenetic analysis was performed through 16 S rRNA gene sequencing. The characteristics of quinoline degradation by different strains were investigated under different pH and temperature conditions. [Results] Based on the phylogenetic analysis,four isolates Q1, Q3, Q7 and Q8 were identified as the genera of Sphingobium, Massilia, Rhodococcus and Dyadobacter, respectively. We demonstrated that 50 mg/L quinoline could be removed within 48 h by all of the strains with varied degradation characteristics. The accumulation of 2-hydroxyquinoline was detected during the cultivation of strain Q1, Q3 and Q8. Notably, the genera of Sphingobium, Massilia and Dyadobacter have not been reported yet on quinoline degradation. [Conclusion] The four quinoline-degrading bacteria isolated from the influent pipeline of bioreactor can provide novel strain resources for decontaminating industrial wastewater containing quinoline, which may contribute to further understanding of the mechanism of quinoline biodegradation.
引文
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[13]Mao YJ,Zhang XJ,Zhang BR,et al.Specific-PCR and denaturing gradient gel electrophoresis assistant isolation of Thauera spp.from a coking wastewater treatment plant[J].Acta Microbiologica Sinica,2008,48(12):1634-1641(in Chinese)毛跃建,张晓君,张宝让,等.专一性PCR和变性梯度胶电泳协助从焦化废水处理装置中分离优势功能菌Thauera属菌株[J].微生物学报,2008,48(12):1634-1641
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[16]Sun QH,Bai YH,Zhao C,et al.Aerobic biodegradation characteristics and metabolic products of quinoline by a Pseudomonas strain[J].Bioresource Technology,2009,100(21):5030-5036
[17]Zhu SN,Liu DQ,Fan L,et al.Isolation,identification and degradation characteristics of a quinoline-degrading bacterium Rhodococcus sp.QL2[J].Environmental Science,2008,29(2):488-493(in Chinese)朱顺妮,刘冬启,樊丽,等.喹啉降解菌Rhodococcus sp.QL2的分离鉴定及降解特性[J].环境科学,2008,29(2):488-493
[18]Wang CR,Zhang MR,Cheng FL,et al.Biodegradation characterization and immobilized strains’potential for quinoline degradation by Brevundimonas sp.K4 isolated from activated sludge of coking wastewater[J].Bioscience,Biotechnology,and Biochemistry,2015,79(1):164-170
[19]Zhong HH,Zhang XJ,Yue SQ,et al.Analysis for metabolic activity of predominant bacterial isolates in a lab-scale denitrifying bioreactor for quinoline-degradation[J].Microbiology China,2010,37(5):645-650(in Chinese)仲汇慧,张晓君,岳思青,等.喹啉废水反硝化反应器中优势菌的代谢功能分析[J].微生物学通报,2010,37(5):645-650
[20]Chen XJ,Wang HJ,Xu JJ,et al.Sphingobium hydrophobicum sp.nov.,a hydrophobic bacterium isolated from electronic-wastecontaminated sediment[J].International Journal of Systematic and Evolutionary Microbiology,2016,66(10):3912-3916
[21]Zhang XY,Peng X,Masai E.Recent advances in Sphingobium sp.SYK-6 for lignin aromatic compounds degradation-a review[J].Acta Microbiologica Sinica,2014,54(8):854-867(in Chinese)张晓琰,彭学,政井英司.木质素芳香族化合物降解菌Sphingobium sp.SYK-6的研究进展[J].微生物学报,2014,54(8):854-867
[22]Lang E,Lapidus A,Chertkov O,et al.Complete genome sequence of Dyadobacter fermentans type strain(NS114T)[J].Standards in Genomic Sciences,2009,1(2):133-140
[23]Wang JL,Han LP,Shi HC,et al.Biodegradation of quinoline by gel immobilized Burkholderia sp.[J].Chemosphere,2001,44(5):1041-1046
[24]Zhu SN,Fan L,Ni JR.Metabolic pathway analysis of two quinoline-degrading bacterial strains[J].China Environmental Science,2008,28(5):456-460(in Chinese)朱顺妮,樊丽,倪晋仁.两株喹啉降解菌代谢途径的分析[J].中国环境科学,2008,28(5):456-460
[25]Bai Q,Yang LH,Li RJ,et al.Accelerating quinoline biodegradation and oxidation with endogenous electron donors[J].Environmental Science&Technology,2015,49(19):11536-11542
[26]Li J,Li WY.Screening of a highly efficient quinoline-degrading strain and its enhanced biotreatment on coking waste water[J].Environmental Science,2015,36(4):1385-1391(in Chinese)李静,李文英.喹啉降解菌筛选及其对焦化废水强化处理[J].环境科学,2015,36(4):1385-1391
[2]Felczak A,Zawadzka K,Lisowska K.Efficient biodegradation of quinolone-Factors determining the process[J].International Biodeterioration&Biodegradation,2014,96:127-134
[3]Qiao L,Wang JL.Biodegradation characteristics of quinoline by Pseudomonas putida[J].Bioresource Technology,2010,101(19):7683-7686
[4]Bai YH,Sun QH,Zhao C,et al.Quinoline biodegradation and its nitrogen transformation pathway by a Pseudomonas sp.strain[J].Biodegradation,2010,21(3):335-344
[5]Tuo BH,Yan JB,Fan BA,et al.Biodegradation characteristics and bioaugmentation potential of a novel quinoline-degrading strain of Bacillus sp.isolated from petroleum-contaminated soil[J].Bioresource Technology,2012,107:55-60
[6]Zhu SN,Liu DQ,Fan L,et al.Degradation of quinoline by Rhodococcus sp.QL2 isolated from activated sludge[J].Journal of Hazardous Materials,2008,160(2/3):289-294
[7]Cui MC,Chen FZ,Fu JM,et al.Microbial metabolism of quinoline by Comamonas sp.[J].World Journal of Microbiology and Biotechnology,2004,20(6):539-543
[8]Wang JL,Wu WZ,Zhao X.Microbial degradation of quinoline:kinetics study with Burkholderia picekttii[J].Biomedical and Environmental Sciences,2004,17(1):21-26
[9]Hong X,Zhang XJ,Meng ZQ,et al.Isolation and characteristics of quinoline degrading bacteria from wastewater denitrifying bioreactor[J].Chinese Journal of Applied&Environmental Biology,2008,14(6):803-808(in Chinese)洪璇,张晓君,孟智奇,等.废水反硝化生物反应器中喹啉降解细菌的分离与特性[J].应用与环境生物学报,2008,14(6):803-808
[10]Deng XQ,Ren Y,Wei CH,et al.Isolation and characterization of Achromobacter sp.DN-06 for degradation of pyridine and quinoline[A]//Proceedings of the 2009 Annual Conference of the Chinese Society of Environmental Sciences(Volume IV)[C]Wuhan:Chinese Society for Environmental Sciences,2009(in Chinese)邓秀琼,任源,韦朝海,等.降解吡啶与喹啉的Achromobacter sp.DN-06筛选、鉴定与降解特性研究[A]//中国环境科学学会2009年学术年会论文集(第4卷)[C].武汉:中国环境科学学会,2009
[11]Johansen SS,Licht D,Arvin E,et al.Metabolic pathways of quinoline,indole and their methylated analogs by Desulfobacterium indolicum(DSM 3383)[J].Applied Microbiology and Biotechnology,1997,47(3):292-300
[12]Licht D,Johansen SS,Arvin E,et al.Transformation of indole and quinoline by Desulfobacterium indolicum(DSM 3383)[J].Applied Microbiology and Biotechnology,1997,47(2):167-172
[13]Mao YJ,Zhang XJ,Zhang BR,et al.Specific-PCR and denaturing gradient gel electrophoresis assistant isolation of Thauera spp.from a coking wastewater treatment plant[J].Acta Microbiologica Sinica,2008,48(12):1634-1641(in Chinese)毛跃建,张晓君,张宝让,等.专一性PCR和变性梯度胶电泳协助从焦化废水处理装置中分离优势功能菌Thauera属菌株[J].微生物学报,2008,48(12):1634-1641
[14]Renshaw MA,Olds BP,Jerde CL,et al.The room temperature preservation of filtered environmental DNA samples and assimilation into a phenol-chloroform-isoamyl alcohol DNAextraction[J].Molecular Ecology Resources,2015,15(1):168-176
[15]Zhang PH,Jia R,Zhang YX,et al.Quinoline-degrading strain Pseudomonas aeruginosa KDQ4 isolated from coking activated sludge is capable of the simultaneous removal of phenol in a dual substrate system[J].Journal of Environmental Science and Health,Part A,2016,51(13):1139-1148
[16]Sun QH,Bai YH,Zhao C,et al.Aerobic biodegradation characteristics and metabolic products of quinoline by a Pseudomonas strain[J].Bioresource Technology,2009,100(21):5030-5036
[17]Zhu SN,Liu DQ,Fan L,et al.Isolation,identification and degradation characteristics of a quinoline-degrading bacterium Rhodococcus sp.QL2[J].Environmental Science,2008,29(2):488-493(in Chinese)朱顺妮,刘冬启,樊丽,等.喹啉降解菌Rhodococcus sp.QL2的分离鉴定及降解特性[J].环境科学,2008,29(2):488-493
[18]Wang CR,Zhang MR,Cheng FL,et al.Biodegradation characterization and immobilized strains’potential for quinoline degradation by Brevundimonas sp.K4 isolated from activated sludge of coking wastewater[J].Bioscience,Biotechnology,and Biochemistry,2015,79(1):164-170
[19]Zhong HH,Zhang XJ,Yue SQ,et al.Analysis for metabolic activity of predominant bacterial isolates in a lab-scale denitrifying bioreactor for quinoline-degradation[J].Microbiology China,2010,37(5):645-650(in Chinese)仲汇慧,张晓君,岳思青,等.喹啉废水反硝化反应器中优势菌的代谢功能分析[J].微生物学通报,2010,37(5):645-650
[20]Chen XJ,Wang HJ,Xu JJ,et al.Sphingobium hydrophobicum sp.nov.,a hydrophobic bacterium isolated from electronic-wastecontaminated sediment[J].International Journal of Systematic and Evolutionary Microbiology,2016,66(10):3912-3916
[21]Zhang XY,Peng X,Masai E.Recent advances in Sphingobium sp.SYK-6 for lignin aromatic compounds degradation-a review[J].Acta Microbiologica Sinica,2014,54(8):854-867(in Chinese)张晓琰,彭学,政井英司.木质素芳香族化合物降解菌Sphingobium sp.SYK-6的研究进展[J].微生物学报,2014,54(8):854-867
[22]Lang E,Lapidus A,Chertkov O,et al.Complete genome sequence of Dyadobacter fermentans type strain(NS114T)[J].Standards in Genomic Sciences,2009,1(2):133-140
[23]Wang JL,Han LP,Shi HC,et al.Biodegradation of quinoline by gel immobilized Burkholderia sp.[J].Chemosphere,2001,44(5):1041-1046
[24]Zhu SN,Fan L,Ni JR.Metabolic pathway analysis of two quinoline-degrading bacterial strains[J].China Environmental Science,2008,28(5):456-460(in Chinese)朱顺妮,樊丽,倪晋仁.两株喹啉降解菌代谢途径的分析[J].中国环境科学,2008,28(5):456-460
[25]Bai Q,Yang LH,Li RJ,et al.Accelerating quinoline biodegradation and oxidation with endogenous electron donors[J].Environmental Science&Technology,2015,49(19):11536-11542
[26]Li J,Li WY.Screening of a highly efficient quinoline-degrading strain and its enhanced biotreatment on coking waste water[J].Environmental Science,2015,36(4):1385-1391(in Chinese)李静,李文英.喹啉降解菌筛选及其对焦化废水强化处理[J].环境科学,2015,36(4):1385-1391
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