温度对硝化杆菌(Nitrobacter)活性动力学影响
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摘要
为探究温度对亚硝酸盐氧化细菌中硝化杆菌属(Nitrobacter)活性动力学影响,本试验采用序批式活性污泥(SBR)反应器,在通过改变系统进水亚硝态氮(NO_2~--N)浓度达到富集Nitrobacter基础上,以富含Nitrobacter污泥为对象(宏基因组物种注释和丰度分析显示Nitrobacter占细菌总数40. 3%),考察不同NO_2~--N浓度梯度下亚硝酸盐氧化过程比亚硝态氮氧化速率(SNi OR)变化规律,基于Monod模型考察系统温度对Nitrobacter活性动力学影响,并进行统计学分析.结果表明,30℃条件下SNi OR达到最大(以N/VSS计),为1. 31 g·(g·d)~(-1).统计学分析结果显示,Monod方程可较好地反映不同温度条件下基质底物浓度对Nitrobacter活性影响.基于菲尔普斯方程计算不同温度区间内温度系数(θ)可知,当系统温度低于25℃或高于30℃时,反应速率随温度变化越敏感.
A sequencing batch reactor( SBR) was operated in this study to investigate the effect of temperature on the kinetics of Nitrobacter activity among nitrite oxidizing bacteria. At the beginning of the experiment,the NO_2~--N concentration in the influent was changed to enrich Nitrobacter. Then,the sludge with enriched Nitrobacter was employed to determine the variation of the specific nitrite oxidation rate( SNi OR) during the nitrite oxidation process in batch tests. Metagenomics species annotation and abundance analysis showed that Nitrobacter accounted for 40. 3% of the total bacterial population. The variation of SNi OR in the nitrite oxidation process was investigated under different NO_2~--N concentrations. The effect of temperature on the kinetics of Nitrobacter was investigated using the Monod model. Furthermore,the kinetics model of the effect of temperature on Nitrobacter activity was fitted for statistical analysis.The results showed that SNi OR reached its maximum at 30℃,which was 1. 31 g·( g·d)~(-1). Statistical analysis showed that the Monod equation could describe the effect of substrate concentration on Nitrobacter activity under different temperature conditions. Calculating the temperature coefficient( θ) in different temperature intervals based on the Phelps equation,showed that when the system temperature is lower than 25℃ or higher than 30℃,the reaction rate is more sensitive to temperature changes.
A sequencing batch reactor( SBR) was operated in this study to investigate the effect of temperature on the kinetics of Nitrobacter activity among nitrite oxidizing bacteria. At the beginning of the experiment,the NO_2~--N concentration in the influent was changed to enrich Nitrobacter. Then,the sludge with enriched Nitrobacter was employed to determine the variation of the specific nitrite oxidation rate( SNi OR) during the nitrite oxidation process in batch tests. Metagenomics species annotation and abundance analysis showed that Nitrobacter accounted for 40. 3% of the total bacterial population. The variation of SNi OR in the nitrite oxidation process was investigated under different NO_2~--N concentrations. The effect of temperature on the kinetics of Nitrobacter was investigated using the Monod model. Furthermore,the kinetics model of the effect of temperature on Nitrobacter activity was fitted for statistical analysis.The results showed that SNi OR reached its maximum at 30℃,which was 1. 31 g·( g·d)~(-1). Statistical analysis showed that the Monod equation could describe the effect of substrate concentration on Nitrobacter activity under different temperature conditions. Calculating the temperature coefficient( θ) in different temperature intervals based on the Phelps equation,showed that when the system temperature is lower than 25℃ or higher than 30℃,the reaction rate is more sensitive to temperature changes.
引文
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[24]Huson D H,Auch A F,Qi J,et al.MEGAN analysis of metagenomic data[J].Genome Research,2007,17(3):377-386.
[25]张自杰,彭永臻.对菲尔普斯(Phelps)公式、温度系数θ的实用价值与理论意义的探讨[J].哈尔滨建筑工程学院学报,1983,(2):71-80.
[2]Peng Y Z,Zhu G B.Biological nitrogen removal with nitrification and denitrification via nitrite pathway[J].Applied Microbiology and Biotechnology,2006,73(1):15-26.
[3]Spieck E,Hartwig C,Cormack I,et al.Selective enrichment and molecular characterization of a previously uncultured Nitrospira-like bacterium from activated sludge[J].Environmental Microbiology,2006,8:405-415.
[4]包鹏,王淑莹,马斌,等.不同溶解氧间歇曝气对亚硝酸盐氧化菌的影响[J].中国环境科学,2016,36(9):2696-2702.Bao P,Wang S Y,Ma B,et al.Effect of dissolve oxygen on the microbial community of the nitrite-oxidizing bacteria in an intermittent aeration reactor[J].China Environmental Science,2016,36(9):2696-2702.
[5]孙洪伟,尤永军,赵华南,等.游离氨对硝化菌活性的抑制及可逆性影响[J].中国环境科学,2015,35(1):95-100.Sun H W,You Y J,Zhao H N,et al.Inhibitory effect of free ammonia on the activity of nitrifying bacteria and recoverability[J].China Environmental Science,2015,35(1):95-100.
[6]GabarróJ,GaniguéR,Gich F,et al.Effect of temperature on AOB activity of a partial nitritation SBR treating landfill leachate with extremely high nitrogen concentration[J].Bioresource Technology,2012,126:283-289.
[7]张宇坤,王淑莹,董怡君,等.游离氨和游离亚硝酸对亚硝态氮氧化菌活性的影响[J].中国环境科学,2014,34(5):1242-1247.Zhang Y K,Wang S Y,Dong Y J,et al.Effect of FA and FNAon activity of nitrite-oxidising bacteria[J].China Environmental Science,2014,34(5):1242-1247.
[8]Madigan M T,Martinko J M,Parker J.Brock biology of microorganisms[M].Upper Saddle River,New Jersey:Prentice Hall,2000.
[9]吴鹏,陆爽君,徐乐中,等.温度对ABR-MBR复合工艺处理生活污水的影响及其微生物群落分析[J].环境科学,2014,35(9):3466-3472.Wu P,Lu S J,Xu L Z,et al.Effects of temperature on combined process of ABR and MBR for domestic sewage treatment and analysis of microbial community[J].Environmental Science,2014,35(9):3466-3472.
[10]Goudar C T,Ganji S M,Pujar B G,et al.Substrate inhibition kinetics of phenol biodegradation[J].Water Environmental Research,2000,72(1):50-55.
[11]Edwards V H.The influence of high substrate concentrations on microbial kinetics[J].Biotechnology and Bioengineering,1970,12(5):679-712.
[12]Andrews J F.A mathematical model for the continuous culture of microorganisms utilizing inhibitory substrates[J].Biotechnology and Bioengineering,1968,10(6):707-723.
[13]Haldane J B S.Enzymes[M].London:Longmans Green and Co,1930.
[14]Aiba S,Shoda M,Nagatani M.Kinetics of product inhibition in alcohol fermentation[J].Biotechnology and Bioengineering,1968,10(6):845-864.
[15]Tessier G.Croissance des populations bactériennes et quantitéd'aliment disponible[J].Reviews Science,1942,80:209-216.
[16]Luong J H T.Generalization of Monod kinetics for analysis of growth data with substrate inhibition[J].Biotechnology and Biochemistry,1987,29(2):242-248.
[17]Han K,Levenspiel O.Extended monod kinetics for substrate,product,and cell inhibition[J].Biotechnology and Bioengineering,1988,32(4):430-437.
[18]Park S W,Bae W.Modeling kinetics of ammonium oxidation and nitrite oxidation under simultaneous inhibition by free ammonia and free nitrous acid[J].Process Biochemistry,2009,44(6):631-640.
[19]Carrera J,Jubany I,Carvallo L,et al.Kinetic models for nitrification inhibition by ammonium and nitrite in a suspended and an immobilised biomass systems[J].Process Biochemistry,2004,39(9):1159-1165.
[20]Ciudad G,Werner A,Bornhardt C,et al.Differential kinetics of ammonia-and nitrite-oxidizing bacteria:A simple kinetic study based on oxygen affinity and proton release during nitrification[J].Process Biochemistry,2006,41(8):1764-1772.
[21]唐崇俭,熊蕾,王云燕,等.高效厌氧氨氧化颗粒污泥的动力学特性[J].环境科学,2013,34(9):3544-3551.Tang C J,Xiong L,Wang Y Y,et al.Kinetic characteristics of high-rate ANAMMOX granules[J].Environmental Science,2013,34(9):3544-3551.
[22]国家环境保护总局.水和废水监测分析方法[M].(第三版).北京:中国环境科学出版社,1997.
[23]Huson D H,Mitra S,Ruscheweyh H J,et al.Integrative analysis of environmental sequences using MEGAN4[J].Genome Research,2011,21(9):1552-1560.
[24]Huson D H,Auch A F,Qi J,et al.MEGAN analysis of metagenomic data[J].Genome Research,2007,17(3):377-386.
[25]张自杰,彭永臻.对菲尔普斯(Phelps)公式、温度系数θ的实用价值与理论意义的探讨[J].哈尔滨建筑工程学院学报,1983,(2):71-80.