球形红细菌降解对硝基酚特性及响应面优化
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摘要
以对硝基酚(PNP)为目标污染物,利用球形红细菌(Rhodobacter sphaeroides)H菌株研究其对PNP的降解特性,通过单因素实验和响应面分析相结合的方法优化降解条件,以提高H菌株对PNP的降解能力。设置不同反应体系证明了H菌株活细胞是降解PNP主体,且在厌氧光照、厌氧黑暗、好氧光照和好氧黑暗四种条件下均能降解PNP。通过单因素实验得出显著影响因素为:PNP初始浓度、pH值和温度,响应面优化后的最优降解条件为:PNP初始浓度为81.01 mg·L~(-1)、pH值8.09和温度30.49℃,PNP降解率的预测值为92.3%,与实际值(91.1%)相差1.2%(<2%),说明预测值可靠。在最优条件下,H菌株的生长和PNP浓度随时间变化关系表明,在H菌株生长的适应期96 h内,PNP浓度从81.01 mg·L~(-1)降低到20.33 mg·L~(-1),降解率为74.9%,指数生长期96～168 h,PNP被快速降解,降解率达到91.1%;同时,拟合了该条件下H菌株降解PNP的一级动力学方程。
With p?nitrophenol(PNP)as the target pollutant,the degradation characteristics of PNP by Rhodobacter sphaeroides H strain were studied. The degradation conditions were optimized by single factor test and response surface analysis,and the degradation ability of H strain to PNP was improved. Different reaction systems have been set up to prove that H strain living cells are the main body of degrading PNP,and can degrade PNP under anaerobic light,anaerobic darkness,aerobic light and aerobic darkness. The single factor experiments show that the significant influencing factors are initial concentration of PNP,pH value and temperature. The optimal degradation conditions after response surface optimization are:initial concentration of PNP is 81.01 mg·L~(-1),pH value is 8.09 and temperature is 30.49 ℃. The predicted value of PNP degradation rate is 92.3%,which is1.2%(<2%)different from the actual value(91.1%). Under the optimum conditions,the relationship between the growth of H strain and the concentration of PNP with time shows that the concentration of PNP decreased from 81.01 mg·L~(-1) to 20.33 mg·L~(-1) within 96 hours of the growth adaptation period of H strain,and the corresponding degradation rate is 74.9%. Then,in the expo?nential growth period of 96-168 hours,PNP is rapidly degraded,and the degradation rate reaches 91.1%. At the same time,the first?order kinetic equation of PNP degradation of H strain under this condition was fitted.
With p?nitrophenol(PNP)as the target pollutant,the degradation characteristics of PNP by Rhodobacter sphaeroides H strain were studied. The degradation conditions were optimized by single factor test and response surface analysis,and the degradation ability of H strain to PNP was improved. Different reaction systems have been set up to prove that H strain living cells are the main body of degrading PNP,and can degrade PNP under anaerobic light,anaerobic darkness,aerobic light and aerobic darkness. The single factor experiments show that the significant influencing factors are initial concentration of PNP,pH value and temperature. The optimal degradation conditions after response surface optimization are:initial concentration of PNP is 81.01 mg·L~(-1),pH value is 8.09 and temperature is 30.49 ℃. The predicted value of PNP degradation rate is 92.3%,which is1.2%(<2%)different from the actual value(91.1%). Under the optimum conditions,the relationship between the growth of H strain and the concentration of PNP with time shows that the concentration of PNP decreased from 81.01 mg·L~(-1) to 20.33 mg·L~(-1) within 96 hours of the growth adaptation period of H strain,and the corresponding degradation rate is 74.9%. Then,in the expo?nential growth period of 96-168 hours,PNP is rapidly degraded,and the degradation rate reaches 91.1%. At the same time,the first?order kinetic equation of PNP degradation of H strain under this condition was fitted.
引文
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[27]郑永良,肖婷,钟玉林,等.一株酚降解菌株的分离鉴定及特性研究[J].湖北农业科学,2010,20(9):2097-2100.ZHENG Liang-yong,XIAO Ting,ZHONG Yu-lin,et al.Isolation and identification of a phenol degra-dation strain and its characteristics analysis[J].Hubei Agricultural Sciences,201 0,20(9):2097-2100.
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[3]郑凤英,钱沙华,李顺兴,等.3,5-二硝基水杨酸表面修饰纳米Ti O_2吸附对硝基苯酚[J].环境科学,2006,27(6):1140-1143.ZHENG Feng-ying,QIAN Sha-hua,LI Shun-xin,et al.Adsorption of p-nitrophenol by nanosized titani-um dioxide surface modified with 3,5-dinitrosalicylic acid[J].Environmental Science,2006,27(6):1140-1143.
[4]万年升,顾继东,黄锦辉,等.Achromobacter xylosoxidans NS12的分离和对硝基苯酚的降解[J].环境科学,2007,28(2):422-426.WAN Nian-sheng,GU Ji-dong,HUANG Jin-hui,et al.Isolation of Achromobacter xylosoxidans NS12 and degradation of nitrophenols[J].Environmental Science,2007,28(2):422-426.
[5]Zhilin Yang,Junxian Chen,Yang Zhou.Understanding the hydrogen transfer mechanism forthe biodegra-dation of 2,4,6-trinitrotoluene catalyzed by pentaerythritol tetranitrate reductase:molecular dynamics si-mulations[J].Phys Chem Chem Phys,2018,20(7):12157-12165.
[6]Suresh R.Subashchandrabose Rhodococcus wratislaviensis strain 9:an efficient p-nitrophenol degader with a great potential for bioremediation[J].Journal of Hazardous Materials,2018,34(7):176-183.
[7]Sahoo NK.Pakshirajan K,Ghosh PK,et al.Batch biodegradation of para-nitrophenol using Arthrobacterchlorophenolicus A6[J].Applied Biochemistry and Biotechnology,2011,1 65(7):1587-1 596.
[8]黄强,张明强.固定化铜绿假单胞菌生物降解对硝基苯酚[J].环境工程技术学报,2012,3(2):247-252.HUANG Qiang,ZHANG Ming-qiang.Biodegradation of p-Nitrophenol by im mobilized cells of Pseudomonas aeruginosa[J].Journal of Environmental Engineering Technology,2012,3(2):247-252.
[9]Bhaswati Chakraborty.Kinetic study of degradation of p-nitrophenol by a mixed bacterial culture and it constituent pure strains[J].Materials Today:Proceedings,201 6,1 0(3):3505-3 524.
[10]李可峰,陈海涛,吴龙飞,等.细菌的光响应及其机制研究进展[J].微生物学通报,201 8,45(7):1574-1 587.LI Ke-feng,CHENG Hai-tao,WU Long-fei,et al.Behavior and mechanism of bacterial response to light illumination[J].Microbiol.China,201 8,45(7):1 574-1 587.
[11]LIANG Fang-nan,BAI Hong-juan,CHAI Chun-jing,et al.Anaerobic biodegradation of 2,4-dinitrotoluene by Rhodobactersphaeroides[J].Microbiology Ch ina,201 6,43(2):279-284.
[12]王玉芬,张肇铭,胡筱敏,等.球形红细菌好氧降解氯代苯研究[J].环境工程学报,2011,5(5):1187-1193.WANG Yu-fen,ZHANG Zhao-ming,HU Xiao-min,et al.Study on aerobic degradation of chlorobenzene by Rhodoba-cter sphaeroides[J].Ch inese Jouinal of Environmental Engineering,2011,5(5):1 187-1 193.
[13]康鹏洲,白红娟,罗征,等.球形红细菌对六价铬的生物还原与三价铬积累[J].国际药学研究杂志,2018,45(4):380-386.KANG Peng-zhou,BAI Hong-juan,LUO Zheng,et al.Biological reduction of hexavalent ch romium and trivalent c-h romium accumu lation by Rhodobacter sphaeroides[J].Journal of International Pharm aceutical Research,2018,45(4):380-386.
[14]红娟,王珊,柴春境,等.球形红细菌降解RDX的动力学及其机理研究[J].火炸药学报,2015,38(6):51-60.BAI Hong-juan,WANG Shan,CHAI Chun-jing,et al.Study on degradation kinetics and mechanism of explosive hexahyd ro-1,3,5-trinitro-1,3,5-triazine(RDX)by Rhodobacter sphaeroides[J].Ch inese Jouinal of Explosives&Propellants,2015,38(6):51-60.
[15]齐永强,王红旗,刘敬奇,等.土壤石油微生物降解影响因子的正交实验分析[J].地球学报,2003,24(3):279-284.QI Yong-qiang,WANG Hong-qi,LIU Jing-qi,et al.Impact of several factors on the bioremediation of oil in soil[J].Acta Geoscientia Sinica,2003,24(3):279-284.
[16]张东升,余丽胜,焦纬洲,等.基于响应面法的超声强化铁碳微电解处理硝基苯废水工艺优化研究[J].含能材料,2018,26(2):178-184.ZHANG Dong-sheng,YU Li-sheng,JIAO Wei-zhou,et al.Treatment of nitrobenzene wastewater via ultrasonic enhanced iron-carbon micro-electrolysis with response surface methodology[J].Ch inese Journal of Energetic Materials(Hanneng Cail-iao),201 8,26(2):1 78-184.
[17]曾茂贵,李颖.正交设计与响应面优化法对瓜蒌桂枝解痉颗粒提取工艺的比较[J].福建中医药大学学报,201 4,24(3):32-36.ZENG Mao-gui,LI Ying.Comparative study on orthogonal design and response surface optimiza-tion used for extraction of gualou guizhi jiejing granules[J].Journal of Fujian University of Tra-ditional Chinese Medicine,2014,24(3):32-36.
[18]姚竹云,张肇铭.几株光合细菌的表型特征及D N A-D N A同源性分析[J].应用与环境生物学报,1996,2(1):84-89.YAO Zhu-yun,ZHANG Zhao-ming.Phenotypic features and DNA-DNA homology analyses of some photosynthetic b-acteria[J].Chinese Journal of Applied and En vironmental Biology,1996,2(1):84-89.
[19]陈正军.黄河兰州段铬还原菌和对硝基酚降解菌的分离筛选及其在微生物燃料电池中的应用研究[D].兰州:兰州大学,201 6.CHEN Zheng-jun.Isolation and screening of strains for chromate reduction and p-nitrophenol degradati-on from the Lanzhou reaches of the Yellow river and their applications in microbial fuel cells[D].Lanzhou:Lanzhou University,201 6.
[20]徐向宏.试验设计与Design-Expert、SPSS应用[M].北京:科学出版社,2010.XU Xiang-hong.Test design and application of Design-Expert and SPSS[M].Beijing:Science Press,2010.
[21]丁丹,胡忠策,金赞芳,等.光合细菌降解废水中对硝基苯酚的研究[J].安徽农业科学,2010,38(1 9):21 9-221.DING Dan,HU Zhong-ce,JIN Zan-fang,et al.Study on degradation of p-nitrophenol in wastewater by photosyntheticbacteria[J].Journal of Anhui Agricultural Sciences,201 0,38(1 9):21 9-221.
[22]胡筱敏,董怡华,李亮,等.光合细菌PSB-1D对2-氯苯酚的降解特性研究[J].环境科学,2010,31(7):1 672-1 678.HU Xiao-min,DONG Yi-hua,LI Liang,et al.Biodegradation characteristics of o-chlorophenol with photosynthetic bacteria PSB-1 D[J].Environmental Science,2010,31(7):1 672-1 678.
[23]董小军,洪青,李恋,等.对硝基苯酚降解菌Pseudomonas sp.PDS-7的降解特性及其降解相关基因的克隆[J].微生物学报,2008,48(1 1):1486-1492.DONG Xiao-jun,HONG Qing,LI Lian,et al.Characterization of a p-nitrophenol degrading bacterium Pseudom-nas sp.PDS-7 and cloning of degradation relevant genes[J].Acta Microbiologica Sinica,2008,48(1 1):1 486-1 492.
[24]尹园,马佳歌,倪春蕾,等.居间驹形氏杆菌发酵大豆糖蜜生产细菌纤维素条件的优化[J].食品科学,201 7,38(1 8):8-1 6.YI Yuan,MA Jia-ge,NI Chun-lei,et al.Optim ization of bacterial cellulose production by fermented soybean molas-ses with Kom aga taeibacter in termedius[J].Acta M icrobiologica Sin ica,2017,38(18):8-16.
[25]刘雪莲.接种量对红景天药渣发酵制有机肥的影响[J].吉林农业,2014,20(22):1 674-1685.LIU Xue-lian.Effect of inoculation amount on the production of organic fertilizer from rhodiola-sachalinensis residues[J].Jilin Agriculture,2014,20(22):1674-1 685.
[26]Samuel M.Bioremediation of p-Nitrophenol by Pseudomonas putida strain[J].Journal of Environmental Health Science and Engineering,2014,12(1):1-8.
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