厌氧微生物降解苯酚过程中胞外蛋白及多糖变化分析
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摘要
为研究厌氧微生物在降解有毒难降解类物质时,胞外多聚物(EPS)的成分变化情况,选取苯酚作为培养基质,按苯酚浓度由低到高(100、300和500 mg/L)分三阶段驯化颗粒污泥。试验过程中,提取对照组(苯酚浓度为零)和三个阶段驯化的颗粒污泥的EPS,测定胞外蛋白质(PN)和胞外多糖(PS)的含量,并对PN和颗粒污泥分别进行三维荧光光谱和电镜扫描(SEM)分析。结果表明:EPS中PN含量变化明显,是抵御苯酚毒性的主要物质;PS含量变化不明显,基本维持在2.6~3.2 mg/g VSS之间,通过其中的负性官能团相互连接构成网状结构,起到维持颗粒污泥形态的作用。通过三维荧光光谱(EEM)分析,细胞分泌的酪氨酸类蛋白质和其他芳香族类蛋白质是厌氧微生物能够有效抵御苯酚毒性的主要原因,且其含量与苯酚浓度呈正相关。SEM照片显示,EPS对污泥结构有影响,PN/PS值增加促进污泥粒径变大,PN减少使污泥颗粒逐渐裂解。
In order to study the change of extracellular polymeric substances( EPS) in the degradation of toxic refractory substances by anaerobic microorganisms,phenol was chosen as the culture medium and the granular sludge was domesticated in three stages according to phenol concentration from low to high( 100 mg/L,300 mg/L and 500 mg/L). The EPS of control group( phenol concentration was zero)and three stages of domesticated granular sludge was extracted,and the content of extracellular protein( PN) and polysaccharide( PS) were determined. Three dimensional molecular fluorescence technique and scanning electron microscopy( SEM) were used to analyze PN and the internal structure of granular sludge respectively. The results showed that the content of PN in EPS changed obviously,which was the main substance to resist the toxicity of phenol. PS content was basically stable,maintaining between 2. 6-3. 2 mg/g VSS. The negative functional groups of PS were connected to each other to form a network structure,which could maintain the shape of granular sludge. Through excitation-emission-matrix( EEM) spectra analysis,the protein of tyrosine and other aromatic species secreted by cells could effectively resist the toxicity of phenol,and the content was positively correlated with the concentration of phenol. SEM pictures showed that EPS affected the sludge structure. The increase of PN/PS value promoted the particle size of sludge,and the decrease of PN caused the sludge particles to break down gradually.
In order to study the change of extracellular polymeric substances( EPS) in the degradation of toxic refractory substances by anaerobic microorganisms,phenol was chosen as the culture medium and the granular sludge was domesticated in three stages according to phenol concentration from low to high( 100 mg/L,300 mg/L and 500 mg/L). The EPS of control group( phenol concentration was zero)and three stages of domesticated granular sludge was extracted,and the content of extracellular protein( PN) and polysaccharide( PS) were determined. Three dimensional molecular fluorescence technique and scanning electron microscopy( SEM) were used to analyze PN and the internal structure of granular sludge respectively. The results showed that the content of PN in EPS changed obviously,which was the main substance to resist the toxicity of phenol. PS content was basically stable,maintaining between 2. 6-3. 2 mg/g VSS. The negative functional groups of PS were connected to each other to form a network structure,which could maintain the shape of granular sludge. Through excitation-emission-matrix( EEM) spectra analysis,the protein of tyrosine and other aromatic species secreted by cells could effectively resist the toxicity of phenol,and the content was positively correlated with the concentration of phenol. SEM pictures showed that EPS affected the sludge structure. The increase of PN/PS value promoted the particle size of sludge,and the decrease of PN caused the sludge particles to break down gradually.
引文
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[3]Puyol D,Monsalvo V M,Mohedano A F,et al.Cosmetic wastewater treatment by upflow anaerobic sludge blanket reactor[J].J Hazard Mater,2011,185:1059-1065.
[4]郁丹,阮文权,邹华,等.厌氧生物法处理苯酚废水[J].食品与生物技术学报,2007,26(5):84-87.Yu Dan,Ruan Wenquan,Zou Hua,et al.Study on phenol wastewater in anaerobic system[J].Journal of Food Science and Biotechnology,2007,26(5):84-87(in Chinese).
[5]Mc Swain B S,Irvine R L,Hausner M,et al.Composition and distribution of extracellular polymeric substances in aerobic flocs and granular sludge[J].Appl Environ Microbiol,2005,71(2):1051-1057.
[6]李硕,彭永臻,王然登.胞外聚合物在污水生物处理中的作用[J].黑龙江大学自然科学学报,2016,33(4):515-520.Li Shuo,Peng Yongzhen,Wang Randeng.The role of extracellular polymers substance in biological treatment of wastewater[J].Journal of Natural Science of Heilongjiang University,2016,33(4):515-520(in Chinese).
[7]李冬,田海成,梁瑜海,等.水质条件对厌氧氨氧化颗粒污泥EPS含量的影响[J].哈尔滨工业大学学报,2017,49(2):6-12.Li Dong,Tian Haicheng,Liang Yuhai,et al.Effect of water quality of influent on the content of extracellular polymeric substances in anammox granule sludge[J].Journal of Harbin Institute of Technology,2017,49(2):6-12(in Chinese).
[8]王淑莹,何岳兰,李夕耀,等.不同活性污泥胞外聚合物提取方法优化[J].北京工业大学学报,2016,42(4):569-576.Wang Shuying,He Yuelan,Li Xiyao,et al.Optimization of extracellular polymeric substance extraction method of different sludge[J].Journal of Beijing University of Technology,2016,42(4):569-576(in Chinese).
[9]徐晓飞,陈健.多糖质量分数测定的研究进展[J].食品科学,2009,30(21):443-448.Xu Xiaofei,Chen Jian.Research progress in methods for quantitative determination of polysaccharides[J].Food Science,2009,30(21):443-448(in Chinese).
[10]虞嘉东,张振家,王欣泽.厌氧颗粒污泥粒径分布的分析测试方法简介[J].工业用水与废水,2004,35(1):57-59.Yu Jiadong,Zhang Zhenjia,Wang Xinze.A brief introduction to the analytical test method for the size distribution of anaerobic granular sludge[J].Industrial Water&Wastewater,2004,35(1):57-59(in Chinese).
[11]马小云,万金泉.苯酚对厌氧颗粒污泥的毒性研究[J].环境科学,2011,32(5):1402-1406.Ma Xiaoyun,Wan Jinquan.Study on toxicity of phenol to anaerobic granular sludge[J].Environmental Science,2011,32(5):1402-1406(in Chinese).
[12]Wang W Y,Liu W Y,Wang L Y,et al.Characteristics and distribution research on extracellular polymer substance extracted from sewage sludge[J].Journal of Environmental Biology,2016,37(2):305-312.
[13]贾学斌,孙云龙,魏金亮,等.厌氧微生物耐受苯酚毒性及降解机理研究[J].哈尔滨商业大学学报:自然科学版,2016,32(5):549-553,562.Jia Xuebin,Sun Yunlong,Wei Jinliang,et al.Anaerobic microorganisms tolerate toxic and mechanism of anaerobic degradation of phenol[J].Journal of Harbin University of Commerce:Natural Sciences Edition,2016,32(5):549-553,562(in Chinese).
[14]Tay J H,Liu Q S,Liu Y.The role of cellular polysaccharides in the formation and stability of aerobic granules[J].Letters in Applied Microbiology,2001,33(3):222-226.
[15]Wang Z P,Liu L L,Yao H,et al.Effects of extracellular polymeric substances on aerobic granulation in sequencing batch reactors[J].Chemosphere,2006,63(10):1728-1735.
[16]Chen W,Westerhoff P,Leenheer J A,et al.Fluorescence excitation-emission matrix regional integration to quantify spectra for dissolved organic matter[J].Environ Sci Technol,2003,37(24):5701-5710.
[17]Ni Bingjie,Yu Hanqing.Microbial products of activated sludge in biological wastewater treatment system:A critical review[J].Critical Reviews in Environmental Science and Technology,2011,42(2):187-223.
[18]Seviour T,Pijuan M,Nicholson T,et al.Understanding the properties of aerobic sludge granules as hydrogels[J].Biotechnol Bioeng,2009,102(5):1483-1493.