固定化镰刀菌降解对氯苯酚废水的研究
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摘要
氯酚类化合物(CPs)作为一类非常重要的工业有机化合物,被广泛应用于许多化工品如染料、颜料、防腐剂、除草剂、杀虫和杀菌剂等的生产中。利用固定化微生物技术处理氯酚废水是环境工程领域的研究热点之一。本文提出了利用镰刀菌处理对氯苯酚(4-CP)废水的方法,研究了镰刀菌降解对氯苯酚的特性和产酶特性。初步研制了一种新型固定化材料-CMC-膨润土凝胶材料,探讨了CMC-膨润土的物化特性,并采用流化床反应器,利用固定化镰刀菌处理对氯苯酚废水,取得了一些有一定价值的研究结果：
1、菌体能以对氯苯酚作为唯一碳源生长,外加碳源可促进对氯苯酚的降解速率。在添加蔗糖浓度1g·L-1、pH6.0、温度30℃条件下,96小时内,100mg·L-1。对氯苯酚的降解率可达100%,游离HJ01菌的最大耐酚量为100mg·L-1。最佳氮源和磷源分别为NH4-N和H2P04-P。4-CP为唯一碳源和外加蔗糖下的降解动力学分别符合Haldane模型和一级动力学方程。
2、镰刀菌HJ01能同时产生氯代邻苯二酚1,2加氧酶(CC120)和氯代邻苯二酚2,3加氧酶(CC230),两种酶皆为诱导酶。CC120的最适宜pH为6.8,温度为50℃,CC230在碱性条件下适应情况较好,且具有较强的耐热性,其最适宜pH和温度分别为8和60℃。两种酶反应动力学符合米氏方程。
3、新型固定化材料CMC-膨润土材料的密度范围为1.00-1.07×103 kg·m-3,含水率90%左右,材料对甲基橙和对氯苯酚都表现出良好的传质性能。
4、不同固定化方法对对氯苯酚降解率比较,CMC-膨润土包埋固定>CBC吸附固定>海藻酸钙固定>游离菌。装置间歇运行时,最佳曝气量为0.8L·min-1。连续进水时,反应器最大耐酚量为100mg·L-1。进水负荷为50mg·L-1,停留时间4h,曝气量0.8 L·min-1条件下,反应器连续运2d,出水稳定,降解率维持在65%左右,固定化材料也基本没有分散破裂的现象,这表明该种材料包埋微生物在废水处理中具有一定的实践意义。不同停留时间下,固定化镰刀菌对4-CP的降解基本符合零级反应动力学。
Chlorophenol are widely distributed due to their common presence in the effluents of many industrial processes, including dye, pigment, preservative,herbicide, insecticide, fungicide industries. The application of immobilization techniques to chlorophenols effluents has been the subject of numerous investigations by environmental workers. In this research, Fusarium HJ01 has been applied to degrade 4-chlorophenol(4-CP). The speciality of HJ01 with potential for 4-chlorophenol degradation and its biodegradation enzyme is investigated. A new carrier, carboxymethylcellulose(CMC)-bentonite gel is studied and partially characterized physically and chemically. The fluidized bed reactor filled with CMC-bentonite gel-immobilized Fusarium HJ01 is constructed and operated to 4-chlorophenol treatment.
The main conclusions of the research are as follows:
1. Fusarium sp.HJ01 could grow using 4-chlorophenol as only carbon resource and energy. Additional carbon can speed the 4-chlorophenol biodegradation.In the condition of 1g·L-1sucrose, pH6,30℃, 100mg·L-1 of 4-chlorophenol is entirely degraded within 96h. The maximum of initial 4-chlorophenol concentration which this strain can endure is 100mg·L-1, when above the value,they are inhibited. the best nitrogen resource and phosphorus resource are NH4-N,H2PO4-P,respectively. The kinetic of 4-CP degradation accord with Haldane model when 4-CP is the sole carbon source and first order equation when sugar is added.
2. This strain can exhibit activity of both chlorocatechol 1,2-dioxygenase (CC12O) and chlorocatechol 2,3-dioxygenase(CC23O) in free cell extracts. The activities of CC12O and CC23O reach the highest value which were 0.609U.mg-1 and 0.125U.mg-1 respectively when use 4-chlorophenol as single carbon resource. When CC12O make function, activity is optimal at pH6.8,50℃, while CC23O has a higher activity in alkalescence condition and is more stable at elevated temperatures when the optimal condition is pH8,60℃. Kinetics of enzyme-catalyzed reactions of CC12O and CC23O accord with The Michaelis-Menten equation.
3. The density of CMC-bentonite gel are 1.00-1.07×103 kg·m-3, water content of the material is about 90%. The material appears a good character of 4-CP and cymene orange mass transportation.
4、In the condition of the same biomass and solid to liquid ratio, the comparison of 4-chlorophenol degradation rate are CMC-bentonite gel-immobilized by embedment> CBC gel-immobilized by absorption> Ca-alginate-immobilized> free cells. In batch-recirculation mode experiment, the optimal air flow rate is 0.8L.min-1. In continuous flow process, the degradation rate obviously fell when the concentration of 4-chlorophenol in influent is larger than 100 mg·L-1. The reactor operated stably in 2 days under these conditions:concentration of 4-chlorophenol in influent is 50 mg·L-1, retention time 4h, air flow rate is 0.8L.min-1,the degradation rate in effluent maintained at about 65%, and the CMC-bentonite gel is not broken up, it shows that this carrier had some practical meaning in wastewater treatment. The kinetic of 4-CP degradation in different retention time accords with zero order equation.
1、菌体能以对氯苯酚作为唯一碳源生长,外加碳源可促进对氯苯酚的降解速率。在添加蔗糖浓度1g·L-1、pH6.0、温度30℃条件下,96小时内,100mg·L-1。对氯苯酚的降解率可达100%,游离HJ01菌的最大耐酚量为100mg·L-1。最佳氮源和磷源分别为NH4-N和H2P04-P。4-CP为唯一碳源和外加蔗糖下的降解动力学分别符合Haldane模型和一级动力学方程。
2、镰刀菌HJ01能同时产生氯代邻苯二酚1,2加氧酶(CC120)和氯代邻苯二酚2,3加氧酶(CC230),两种酶皆为诱导酶。CC120的最适宜pH为6.8,温度为50℃,CC230在碱性条件下适应情况较好,且具有较强的耐热性,其最适宜pH和温度分别为8和60℃。两种酶反应动力学符合米氏方程。
3、新型固定化材料CMC-膨润土材料的密度范围为1.00-1.07×103 kg·m-3,含水率90%左右,材料对甲基橙和对氯苯酚都表现出良好的传质性能。
4、不同固定化方法对对氯苯酚降解率比较,CMC-膨润土包埋固定>CBC吸附固定>海藻酸钙固定>游离菌。装置间歇运行时,最佳曝气量为0.8L·min-1。连续进水时,反应器最大耐酚量为100mg·L-1。进水负荷为50mg·L-1,停留时间4h,曝气量0.8 L·min-1条件下,反应器连续运2d,出水稳定,降解率维持在65%左右,固定化材料也基本没有分散破裂的现象,这表明该种材料包埋微生物在废水处理中具有一定的实践意义。不同停留时间下,固定化镰刀菌对4-CP的降解基本符合零级反应动力学。
Chlorophenol are widely distributed due to their common presence in the effluents of many industrial processes, including dye, pigment, preservative,herbicide, insecticide, fungicide industries. The application of immobilization techniques to chlorophenols effluents has been the subject of numerous investigations by environmental workers. In this research, Fusarium HJ01 has been applied to degrade 4-chlorophenol(4-CP). The speciality of HJ01 with potential for 4-chlorophenol degradation and its biodegradation enzyme is investigated. A new carrier, carboxymethylcellulose(CMC)-bentonite gel is studied and partially characterized physically and chemically. The fluidized bed reactor filled with CMC-bentonite gel-immobilized Fusarium HJ01 is constructed and operated to 4-chlorophenol treatment.
The main conclusions of the research are as follows:
1. Fusarium sp.HJ01 could grow using 4-chlorophenol as only carbon resource and energy. Additional carbon can speed the 4-chlorophenol biodegradation.In the condition of 1g·L-1sucrose, pH6,30℃, 100mg·L-1 of 4-chlorophenol is entirely degraded within 96h. The maximum of initial 4-chlorophenol concentration which this strain can endure is 100mg·L-1, when above the value,they are inhibited. the best nitrogen resource and phosphorus resource are NH4-N,H2PO4-P,respectively. The kinetic of 4-CP degradation accord with Haldane model when 4-CP is the sole carbon source and first order equation when sugar is added.
2. This strain can exhibit activity of both chlorocatechol 1,2-dioxygenase (CC12O) and chlorocatechol 2,3-dioxygenase(CC23O) in free cell extracts. The activities of CC12O and CC23O reach the highest value which were 0.609U.mg-1 and 0.125U.mg-1 respectively when use 4-chlorophenol as single carbon resource. When CC12O make function, activity is optimal at pH6.8,50℃, while CC23O has a higher activity in alkalescence condition and is more stable at elevated temperatures when the optimal condition is pH8,60℃. Kinetics of enzyme-catalyzed reactions of CC12O and CC23O accord with The Michaelis-Menten equation.
3. The density of CMC-bentonite gel are 1.00-1.07×103 kg·m-3, water content of the material is about 90%. The material appears a good character of 4-CP and cymene orange mass transportation.
4、In the condition of the same biomass and solid to liquid ratio, the comparison of 4-chlorophenol degradation rate are CMC-bentonite gel-immobilized by embedment> CBC gel-immobilized by absorption> Ca-alginate-immobilized> free cells. In batch-recirculation mode experiment, the optimal air flow rate is 0.8L.min-1. In continuous flow process, the degradation rate obviously fell when the concentration of 4-chlorophenol in influent is larger than 100 mg·L-1. The reactor operated stably in 2 days under these conditions:concentration of 4-chlorophenol in influent is 50 mg·L-1, retention time 4h, air flow rate is 0.8L.min-1,the degradation rate in effluent maintained at about 65%, and the CMC-bentonite gel is not broken up, it shows that this carrier had some practical meaning in wastewater treatment. The kinetic of 4-CP degradation in different retention time accords with zero order equation.
引文
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