嗜盐菌群对酸性金黄G的脱色特性和机理
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摘要
【目的】为了获得能够在高盐环境下脱色偶氮染料的嗜盐菌群及其降解机理。【方法】采用富集驯化的方法获得一个嗜盐菌群,采用Illumina HiSeq2500测序平台对其群落结构进行测定;采用分光光度法测定了其降解特性;采用GC-MS和红外图谱分析了其降解机理;采用微核实验的方法比较了偶氮染料降解前后的毒性。【结果】该菌群在10%的盐度下,使100mg/L的酸性金黄G在8h内脱色。菌群主要由Zobellella、Rheinheimera、Exiguobacterium和Marinobacterium组成。最适宜的脱色条件是:pH=6,酵母粉为碳源,蛋白胨或硝酸钾作为氮源,盐度为1%–10%。酸性金黄G降解产物的毒性比降解前降低。酸性金黄G主要的降解产物是对氨基二苯胺和二苯胺。此外,该菌群还能使酸性大红GR和直接湖蓝5B等多种偶氮染料脱色,具有较好的脱色广谱性。【结论】获得了快速降解偶氮染料的嗜盐菌群及降解机理,为该嗜盐菌群应用于高盐印染废水的处理提供菌种资源和理论支持。
[Objective] In order to obtain highly efficient bacterial consortium that can rapidly decolorize azo dyes in high salinity environment. [Methods] A halophilic bacterial consortium was obtained by enrichment, and its community structure was determined by Illumina HiSeq2500 sequencing platform. The degradation characteristics were determined by spectrophotometry. The degradation mechanism was analyzed by GC-MS and Fourier Transform infrared spectroscopy. The toxicity of azo dye and degradation product were compared by micronuclear test. [Results] At 10% salinity, the bacterial consortium decolorizes methanil yellow G completely in 8 h. The bacterial consortium was composed of Zobellella, Rheinheimera, Exiguobacterium and Marinobacterium. At pH 6.0, with yeast powder as carbon sources, peptone or potassium nitrate as nitrogen sources and 1% NaCl were the optimal decolorization conditions. Decolorization metabolites of methanil yellow G were less toxic than original dye. The main degradation products of methanil yellow G were p-aminodiphenylamine and diphenylamine. In addition, the bacterial consortium also decolorized various azo dyes, such as acid magenta GR and direct lake blue 5 B. [Conclusion] A halophilic bacterial consortium for fast degradation of azo dyes was obtained, with the application potential to treat high-salt printing and dyeing wastewater.
[Objective] In order to obtain highly efficient bacterial consortium that can rapidly decolorize azo dyes in high salinity environment. [Methods] A halophilic bacterial consortium was obtained by enrichment, and its community structure was determined by Illumina HiSeq2500 sequencing platform. The degradation characteristics were determined by spectrophotometry. The degradation mechanism was analyzed by GC-MS and Fourier Transform infrared spectroscopy. The toxicity of azo dye and degradation product were compared by micronuclear test. [Results] At 10% salinity, the bacterial consortium decolorizes methanil yellow G completely in 8 h. The bacterial consortium was composed of Zobellella, Rheinheimera, Exiguobacterium and Marinobacterium. At pH 6.0, with yeast powder as carbon sources, peptone or potassium nitrate as nitrogen sources and 1% NaCl were the optimal decolorization conditions. Decolorization metabolites of methanil yellow G were less toxic than original dye. The main degradation products of methanil yellow G were p-aminodiphenylamine and diphenylamine. In addition, the bacterial consortium also decolorized various azo dyes, such as acid magenta GR and direct lake blue 5 B. [Conclusion] A halophilic bacterial consortium for fast degradation of azo dyes was obtained, with the application potential to treat high-salt printing and dyeing wastewater.
引文
[1]Hua L,Xie JK,Zhu C,Ma HR,Peng XY.Advance in microbial decolorization and degradation of azo dyes.Journal of Northwest A&F University(Natural Science Edition),2015,43(10):103-113.(in Chinese)花莉,解井坤,朱超,马宏瑞,彭香玉.微生物对偶氮染料脱色降解的研究进展.西北农林科技大学学报(自然科学版),2015,43(10):103-113.
[2]Lu J,Yu ZS,Zhang HX.Research progress in the microbial degradation of azo dyes.Industrial Water Treatment,2013,33(1):15-19.(in Chinese)卢婧,余志晟,张洪勋.微生物降解偶氮染料的研究进展.工业水处理,2013,33(1):15-19.
[3]Bhatia D,Sharma NR,Singh J,Kanwar RS.Biological methods for textile dye removal from wastewater:A review.Critical Reviews in Environmental Science and Technology,2017,47(19):1836-1876.
[4]Vikrant K,Giri BS,Raza N,Roy K,Kim KH,Rai BN,Singh RS.Recent advancements in bioremediation of dye:Current status and challenges.Bioresource Technology,2018,253:355-367.
[5]He HJ,Chen YJ,Li X,Cheng Y,Yang CP,Zeng GM.Influence of salinity on microorganisms in activated sludge processes:a review.International Biodeterioration&Biodegradation,2017,119:520-527.
[6]Wei N,Zhang QQ,Du ZJ,Zhang DM.Isolation and identification of halophilic bacteria strain Halomonas elogata and its application on degradation of an azo dye Reactive Blue BRF.Acta Scientiae Circumstantiae,2012,32(9):2091-2096.(in Chinese)韦娜,张前前,杜宗军,张栋梅.极端嗜盐菌Halomonas elogata的分离鉴定及其降解偶氮染料活性兰BRF的条件优化研究.环境科学学报,2012,32(9):2091-2096.
[7]Patel VR,Bhatt N.Isolation,development and identification of salt-tolerant bacterial consortium from crude-oil-contaminated soil for degradation of di-azo dye Reactive Blue 220.Water Science and Technology,2015,72(2):311-321.
[8]Guo G,Wang YH,Tian F,Ding KQ,Xu Y,Chai J,Liu T,Jia YS,Zhang L.Enrichment and decoloring capability of an acid brilliant scarlet GR decoloring halophilic bacterial consortium.Chinese Journal of Environmental Engineering,2016,10(2):631-636.(in Chinese)郭光,王艳华,田芳,丁克强,徐瑶,柴晋,刘婷,贾永胜,张丽.酸性大红嗜盐菌群的富集及脱色效果.环境工程学报,2016,10(2):631-636.
[9]Guo G,He F,Tian F,Huang Y,Wang H.Effect of salt contents on enzymatic activities and halophilic microbial community structure during phenanthrene degradation.International Biodeterioration&Biodegradation,2016,110:8-15.
[10]Guo G,Tian F,Liu Y,Liu TF,Ding KQ,Liu C.Decoloring acid scarlet GR by a moderately halophilic bacterial consortium.Microbiology China,2017,44(11):2567-2574.(in Chinese)郭光,田芳,刘妍,刘廷凤,丁克强,刘翀.中度嗜盐菌群对酸性大红GR的脱色性能研究.微生物学通报,2017,44(11):2567-2574.
[11]Xu XT,Lu P,Wu D,Li XK.Isolation,characterization and toxicity of a malachite green degradation bacterial strain.Microbiology China,2017,44(8):1817-1824.(in Chinese)徐晓婷,卢平,吴丹,李旭凯.孔雀石绿脱色菌的分离鉴定及染料脱色后毒性研究.微生物学通报,2017,44(8):1817-1824.
[12]Xie XH,Fan FX,Yuan XW,Liu N,Liu JS.Isolation of high performance bacterial consortium FF and its decolorization effects on reactive black 5.Journal of Donghua University(Natural Science),2013,39(6):802-813.(in Chinese)谢学辉,范凤霞,袁学武,刘娜,柳建设.高效混合菌群FF的筛选及其对活性黑5的脱色作用.东华大学学报(自然科学版),2013,39(6):802-813.
[13]Song L,Shao YF,Ning SX,Tan L.Performance of a newly isolated salt-tolerant yeast strain Pichia occidentalis G1 for degrading and detoxifying azo dyes.Bioresource Technology,2017,233:21-29
[14]Bai J,Chen L,Huang X,Hu CH,Zhao YG,Li KR.Isolation and nitrogen removal characteristics of salt-tolerant heterotrophic nitrification and aerobic denitrification bacteria Zobellella sp.B307.Environmental Science,2018,39(10):4793-4801.(in Chinese)白洁,陈琳,黄潇,胡春辉,赵阳国,李岿然.1株耐盐异养硝化-好氧反硝化菌Zobellella sp.B307的分离及脱氮特性.环境科学,2018,39(10):4793-4801.
[15]Lee DW,Lee H,Kwon BO,Khim JS,Yim UH,Park H,Park B,Choi IG,Kim BS,Kim JJ.Zobellella maritima sp.nov.,a polycyclic aromatic hydrocarbon-degrading bacterium,isolated from beach sediment.International Journal of Systematic and Evolutionary Microbiology,2018,68(7):2279-2284.
[16]Li XY,Wang LP,Lai QL,Li DF,Shao ZZ.Preliminary study on aerobic denitrification characteristics of Zobellella sp.F13-1.Journal of Applied Oceanography,2016,35(1):122-129.(in Chinese)李小义,王丽萍,赖其良,李登峰,邵宗泽.一株卓贝尔氏菌F13-1好氧反硝化特性及其反硝化基因的初步研究.应用海洋学学报,2016,35(1):122-129.
[17]Zhang Y,Shi P,Ma J.Exiguobacterium spp.and their applications in environmental remediation.Chinese Journal of Applied and Environmental Biology,2013,19(5):898-904.(in Chinese)张莹,石萍,马炯.微小杆菌Exiguobacterium spp.及其环境应用研究进展.应用与环境生物学报,2013,19(5):898-904.
[18]Tan L,Qu YY,Zhou JT,Li A.Decolorization of azo dyes by a newly isolated Exiguobacterium sp.strain TL under high salt condition.Journal of Biotechnology,2008,136 Suppl:S688.
[19]Dhanve RS,Shedbalkar UU,Jadhav JP.Biodegradation of diazo reactive dye Navy blue HE2R(Reactive blue 172)by an isolated Exiguobacterium sp.RD3.Biotechnology and Bioprocess Engineering,2008,13(1):53-60.
[20]Imran M,Arshad M,Negm F,Khalid A,Shaharoona B,Hussain S,Nadeem SM,Crowley DE.Yeast extract promotes decolorization of azo dyes by stimulating azoreductase activity in Shewanella sp.strain IFN4.Ecotoxicology and Environmental Safety,2016,124:42-49.
[21]Khan R,Bhawana P,Fulekar MH.Microbial decolorization and degradation of synthetic dyes:a review.Reviews in Environmental Science and Bio/Technology,2013,12(1):75-97.
[22]Saratale RG,Saratale GD,Chang JS,Govindwar SP.Bacterial decolorization and degradation of azo dyes:A review.Journal of the Taiwan Institute of Chemical Engineers,2011,42(1):138-157.
[23]Xiang XM,Wang XK,Zhou JT,Wang R,Qu BC.Decolorization of reactive X-3B red dye by salt-tolerant bacterium GTY.Environmental Science&Technology,2010,33(2):31-34.(in Chinese)项学敏,王晓坤,周集体,王刃,曲宝成.耐盐菌GTY对偶氮染料活性艳红X-3B的脱色研究.环境科学与技术,2010,33(2):31-34.
[24]Dai SG,Song WH,Li T,Zhuang YY.Study on azo dyes structure-biodegradability relationships.Advances in Environmental Science,1996,4(6):1-9.(in Chinese)戴树桂,宋文华,李彤,庄源益.偶氮染料结构与其生物降解性关系研究进展.环境科学进展,1996,4(6):1-9.
[25]Arun Prasad AS,Satyanarayana VSV,Bhaskara Rao KV.Biotransformation of Direct Blue 1 by a moderately halophilic bacterium Marinobacter sp.strain HBRA and toxicity assessment of degraded metabolites.Journal of Hazardous Materials,2013,262:674-684.
[26]Chen Y,Feng LL,Li HG,Wang YX,Chen GT,Zhang QH.Biodegradation and detoxification of Direct Black G textile dye by a newly isolated thermophilic microflora.Bioresource Technology,2018,250:650-657.
[2]Lu J,Yu ZS,Zhang HX.Research progress in the microbial degradation of azo dyes.Industrial Water Treatment,2013,33(1):15-19.(in Chinese)卢婧,余志晟,张洪勋.微生物降解偶氮染料的研究进展.工业水处理,2013,33(1):15-19.
[3]Bhatia D,Sharma NR,Singh J,Kanwar RS.Biological methods for textile dye removal from wastewater:A review.Critical Reviews in Environmental Science and Technology,2017,47(19):1836-1876.
[4]Vikrant K,Giri BS,Raza N,Roy K,Kim KH,Rai BN,Singh RS.Recent advancements in bioremediation of dye:Current status and challenges.Bioresource Technology,2018,253:355-367.
[5]He HJ,Chen YJ,Li X,Cheng Y,Yang CP,Zeng GM.Influence of salinity on microorganisms in activated sludge processes:a review.International Biodeterioration&Biodegradation,2017,119:520-527.
[6]Wei N,Zhang QQ,Du ZJ,Zhang DM.Isolation and identification of halophilic bacteria strain Halomonas elogata and its application on degradation of an azo dye Reactive Blue BRF.Acta Scientiae Circumstantiae,2012,32(9):2091-2096.(in Chinese)韦娜,张前前,杜宗军,张栋梅.极端嗜盐菌Halomonas elogata的分离鉴定及其降解偶氮染料活性兰BRF的条件优化研究.环境科学学报,2012,32(9):2091-2096.
[7]Patel VR,Bhatt N.Isolation,development and identification of salt-tolerant bacterial consortium from crude-oil-contaminated soil for degradation of di-azo dye Reactive Blue 220.Water Science and Technology,2015,72(2):311-321.
[8]Guo G,Wang YH,Tian F,Ding KQ,Xu Y,Chai J,Liu T,Jia YS,Zhang L.Enrichment and decoloring capability of an acid brilliant scarlet GR decoloring halophilic bacterial consortium.Chinese Journal of Environmental Engineering,2016,10(2):631-636.(in Chinese)郭光,王艳华,田芳,丁克强,徐瑶,柴晋,刘婷,贾永胜,张丽.酸性大红嗜盐菌群的富集及脱色效果.环境工程学报,2016,10(2):631-636.
[9]Guo G,He F,Tian F,Huang Y,Wang H.Effect of salt contents on enzymatic activities and halophilic microbial community structure during phenanthrene degradation.International Biodeterioration&Biodegradation,2016,110:8-15.
[10]Guo G,Tian F,Liu Y,Liu TF,Ding KQ,Liu C.Decoloring acid scarlet GR by a moderately halophilic bacterial consortium.Microbiology China,2017,44(11):2567-2574.(in Chinese)郭光,田芳,刘妍,刘廷凤,丁克强,刘翀.中度嗜盐菌群对酸性大红GR的脱色性能研究.微生物学通报,2017,44(11):2567-2574.
[11]Xu XT,Lu P,Wu D,Li XK.Isolation,characterization and toxicity of a malachite green degradation bacterial strain.Microbiology China,2017,44(8):1817-1824.(in Chinese)徐晓婷,卢平,吴丹,李旭凯.孔雀石绿脱色菌的分离鉴定及染料脱色后毒性研究.微生物学通报,2017,44(8):1817-1824.
[12]Xie XH,Fan FX,Yuan XW,Liu N,Liu JS.Isolation of high performance bacterial consortium FF and its decolorization effects on reactive black 5.Journal of Donghua University(Natural Science),2013,39(6):802-813.(in Chinese)谢学辉,范凤霞,袁学武,刘娜,柳建设.高效混合菌群FF的筛选及其对活性黑5的脱色作用.东华大学学报(自然科学版),2013,39(6):802-813.
[13]Song L,Shao YF,Ning SX,Tan L.Performance of a newly isolated salt-tolerant yeast strain Pichia occidentalis G1 for degrading and detoxifying azo dyes.Bioresource Technology,2017,233:21-29
[14]Bai J,Chen L,Huang X,Hu CH,Zhao YG,Li KR.Isolation and nitrogen removal characteristics of salt-tolerant heterotrophic nitrification and aerobic denitrification bacteria Zobellella sp.B307.Environmental Science,2018,39(10):4793-4801.(in Chinese)白洁,陈琳,黄潇,胡春辉,赵阳国,李岿然.1株耐盐异养硝化-好氧反硝化菌Zobellella sp.B307的分离及脱氮特性.环境科学,2018,39(10):4793-4801.
[15]Lee DW,Lee H,Kwon BO,Khim JS,Yim UH,Park H,Park B,Choi IG,Kim BS,Kim JJ.Zobellella maritima sp.nov.,a polycyclic aromatic hydrocarbon-degrading bacterium,isolated from beach sediment.International Journal of Systematic and Evolutionary Microbiology,2018,68(7):2279-2284.
[16]Li XY,Wang LP,Lai QL,Li DF,Shao ZZ.Preliminary study on aerobic denitrification characteristics of Zobellella sp.F13-1.Journal of Applied Oceanography,2016,35(1):122-129.(in Chinese)李小义,王丽萍,赖其良,李登峰,邵宗泽.一株卓贝尔氏菌F13-1好氧反硝化特性及其反硝化基因的初步研究.应用海洋学学报,2016,35(1):122-129.
[17]Zhang Y,Shi P,Ma J.Exiguobacterium spp.and their applications in environmental remediation.Chinese Journal of Applied and Environmental Biology,2013,19(5):898-904.(in Chinese)张莹,石萍,马炯.微小杆菌Exiguobacterium spp.及其环境应用研究进展.应用与环境生物学报,2013,19(5):898-904.
[18]Tan L,Qu YY,Zhou JT,Li A.Decolorization of azo dyes by a newly isolated Exiguobacterium sp.strain TL under high salt condition.Journal of Biotechnology,2008,136 Suppl:S688.
[19]Dhanve RS,Shedbalkar UU,Jadhav JP.Biodegradation of diazo reactive dye Navy blue HE2R(Reactive blue 172)by an isolated Exiguobacterium sp.RD3.Biotechnology and Bioprocess Engineering,2008,13(1):53-60.
[20]Imran M,Arshad M,Negm F,Khalid A,Shaharoona B,Hussain S,Nadeem SM,Crowley DE.Yeast extract promotes decolorization of azo dyes by stimulating azoreductase activity in Shewanella sp.strain IFN4.Ecotoxicology and Environmental Safety,2016,124:42-49.
[21]Khan R,Bhawana P,Fulekar MH.Microbial decolorization and degradation of synthetic dyes:a review.Reviews in Environmental Science and Bio/Technology,2013,12(1):75-97.
[22]Saratale RG,Saratale GD,Chang JS,Govindwar SP.Bacterial decolorization and degradation of azo dyes:A review.Journal of the Taiwan Institute of Chemical Engineers,2011,42(1):138-157.
[23]Xiang XM,Wang XK,Zhou JT,Wang R,Qu BC.Decolorization of reactive X-3B red dye by salt-tolerant bacterium GTY.Environmental Science&Technology,2010,33(2):31-34.(in Chinese)项学敏,王晓坤,周集体,王刃,曲宝成.耐盐菌GTY对偶氮染料活性艳红X-3B的脱色研究.环境科学与技术,2010,33(2):31-34.
[24]Dai SG,Song WH,Li T,Zhuang YY.Study on azo dyes structure-biodegradability relationships.Advances in Environmental Science,1996,4(6):1-9.(in Chinese)戴树桂,宋文华,李彤,庄源益.偶氮染料结构与其生物降解性关系研究进展.环境科学进展,1996,4(6):1-9.
[25]Arun Prasad AS,Satyanarayana VSV,Bhaskara Rao KV.Biotransformation of Direct Blue 1 by a moderately halophilic bacterium Marinobacter sp.strain HBRA and toxicity assessment of degraded metabolites.Journal of Hazardous Materials,2013,262:674-684.
[26]Chen Y,Feng LL,Li HG,Wang YX,Chen GT,Zhang QH.Biodegradation and detoxification of Direct Black G textile dye by a newly isolated thermophilic microflora.Bioresource Technology,2018,250:650-657.