低C/N比异养硝化-好氧反硝化菌筛选及硝化特性
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摘要
针对生物法处理低C/N比废水存在碳源不足、脱氮效率不高问题,从石化废水处理厂活性污泥中分离得到一株低C/N比异养硝化-好氧反硝化菌株WUST-7。通过形态学观察、生理生化试验和16S rDNA序列分析,鉴定其为假单胞菌属(Pseudomonas sp.)。通过单因素实验,考察碳源种类、培养温度、初始pH和摇床转速对菌株硝化性能的影响,确定最优异养硝化培养条件为:丁二酸钠为碳源、培养温度30～35℃、初始pH8.0～9.0、摇床转速150～200r/min。在最优异养硝化条件下培养9h,可将初始浓度为107.52mg/L的氨氮去除90.64%,并且在整个培养过程中没有亚硝酸盐氮的积累,硝酸盐氮含量也始终低于3.5mg/L,总氮的去除率达88.63%。实验结果表明,菌株WUST-7在利用氨氮进行硝化反应的同时,还可以利用硝酸盐氮进行反硝化,具有良好的同步硝化反硝化潜能。
In view of the problems of insufficient carbon source and low nitrogen removal efficiency for biological treatment of low C/N ratio wastewater, a heterotrophic nitrification-aerobic denitrification strain WUST-7 with low C/N ratio was isolated from the activated sludge of petrochemical wastewater treatment plant. Pseudomonas sp. was identified by morphological observation, physiological and biochemical tests and 16 S rDNA sequence analysis. The effects of carbon source type, culture temperature, initial pH and rotation speed on the nitrification characteristics were investigated by single factor experiments. The optimal conditions for heterotrophic nitrification were determined as follows: sodium succinate as carbon source, culture temperature 30-35℃, initial pH 8.0-9.0, rotation speed 150-200 r/min. Under the optimal culture conditions for 9 h, 90.64 % of ammonia nitrogen with initial concentration of 107.52 mg/L could be removed there was no accumulation of nitrite nitrogen and the nitrate nitrogen content was always lower than 3.5 mg/L in the whole process. The removal rate of total nitrogen is up to 88.63%. The experimental results showed that the strain WUST-7 had good simultaneous nitrification and denitrification potential when it used ammonia nitrogen for nitrification and nitrate nitrogen for denitrification.
In view of the problems of insufficient carbon source and low nitrogen removal efficiency for biological treatment of low C/N ratio wastewater, a heterotrophic nitrification-aerobic denitrification strain WUST-7 with low C/N ratio was isolated from the activated sludge of petrochemical wastewater treatment plant. Pseudomonas sp. was identified by morphological observation, physiological and biochemical tests and 16 S rDNA sequence analysis. The effects of carbon source type, culture temperature, initial pH and rotation speed on the nitrification characteristics were investigated by single factor experiments. The optimal conditions for heterotrophic nitrification were determined as follows: sodium succinate as carbon source, culture temperature 30-35℃, initial pH 8.0-9.0, rotation speed 150-200 r/min. Under the optimal culture conditions for 9 h, 90.64 % of ammonia nitrogen with initial concentration of 107.52 mg/L could be removed there was no accumulation of nitrite nitrogen and the nitrate nitrogen content was always lower than 3.5 mg/L in the whole process. The removal rate of total nitrogen is up to 88.63%. The experimental results showed that the strain WUST-7 had good simultaneous nitrification and denitrification potential when it used ammonia nitrogen for nitrification and nitrate nitrogen for denitrification.
引文
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[4]LIU Y X,WANG Y,LI Y,et al.Nitrogen removal characteristics of heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis C16[J].Chinese Journal of Chemical Engineering,2015,23(5):827-834.
[5]GE Q L,YUE X P,WANG G Y.Simultaneous heterotrophic nitrification and aerobic denitrification at high initial phenol concentration by isolated bacterium Diaphorobacter sp.PD-7[J].Chinese Journal of Chemical Engineering,2015,23(5):835-841.
[6]REN Y X,YANG L,LIANG X.The characteristics of a novel heterotrophic nitrifying and aerobic denitrifying bacterium,Acinetobacter junii YB[J].Bioresource Technology,2014,171:1-9.
[7]CHEN M X,WANG W C,FENG Y,et al.Impact resistance of different factors on ammonia removal by heterotrophic nitrificationaerobic denitrification bacterium Aeromonas sp.HN-02[J].Bioresource Technology,2014,167:456-461.
[8]HE T X,LI Z L,SUN Q,et al.Heterotrophic nitrification and aerobic denitrification by Pseudomonas tolaasii Y-11 without nitrite accumulation during nitrogen conversion[J].Bioresource Technology,2016,200:493-499.
[9]武文丽,颜家保,陈佩,等.炼油废水中好氧反硝化菌的筛选及降解特性[J].化工进展,2016,35(5):1524-1528.WU W L,YAN J B,CHEN P,et al.Screening of an aerobic denitrifier from refinery wastewater and its characteristics[J].Chemical Industry and Engineering Progress,2016,35(5):1524-1528.
[10]乔楠,陈瑞佳,于大禹.硅藻土负载异养硝化-好氧反硝化菌的脱氮性能[J].化工进展,2015,34(5):1459-1465.QIAO N,CHEN R J,YU D Y.Study on the denitrification characteristics of heterotrophic nitrification-aerobic denitrifier loaded on diatomite[J].Chemical Industry and Engineering Progress,2015,34(5):1459-1465.
[11]彭永臻,马斌.低C/N比条件下高效生物脱氮策略分析[J].环境科学学报,2009,29(2):225-230.PENG Y Z,MA B.Review of biological nitrogen removal enhancement technologies and processes under low C/N ratio[J].Acta Scientiae Circumstantiae,2009,29(2):225-230.
[12]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.DONG X Z,CAI M Y.Common bacterial system identification manual[M].Beijing:Science Press,2001.
[13]国家环境保护总局.水和废水监测分析方法[M].4版.北京:中国环境科学出版社,2002.State Environmental Protection Administration of China.Water and wastewater analysis methods[M].4th ed.Beijing:China Environmental Science Press,2002.
[14]YANG L,REN Y X,ZHAO S Q,et al.Isolation and characterization of three heterotrophic nitrifying-aerobic denitrifying bacteria from a sequencing batch reactor[J].Annals of Microbiology,2015,66(2):737-747.
[15]HUANG F,PAN L Q,LV N,et al.Characterization of novel Bacillus strain N31 from mariculture water capable of halophilic heterotrophic nitrification-aerobic denitrification[J].Journal of Bioscience&Bioengineering,2017,124(5):564-571.
[16]张小燕.新金分枝杆菌ZJUVN-08转化植物甾醇合成雄甾烯二酮的研究[D].杭州:浙江大学,2013.ZHANG X Y.Study on synthesis of androstenedione from phytosterol transformed by Mycobacterium neoaurum.ZJUVN-08[D].Hangzhou:Zhejiang University,2013.
[17]XU Y,HE T X,LI Z L,et al.Nitrogen removal characteristics of Pseudomonas putida Y-9 capable of heterotrophic nitrification and aerobic denitrification at low temperature[J].BioMed Research International,2017,2017:1429018.