甲醇和乙醇微生物传感体系的研究
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摘要
第一章:简述了生物传感器及微生物传感器的工作原理、组成、分类及特性。并简单介绍微生物传感器的研究进展。着重对微生物细胞固定化的方法和载体,以及近年来出现的新型纳米材料碳纳米管在生物传感器方面的应用进行简要总结。
第二章:通过对碳纳米管的荧光光谱、红外光谱的分析,考察碳纳米管是否被功能化。采用溶胶—凝胶包埋法,制备固定化小球,就微生物细胞不同固定化方法和载体进行实验比较。对影响溶胶—凝胶固定化的添加剂二氧化硅(SiO_2)、碳酸钙(CaCO_3)、海藻酸钠等浓度进行了考察,得出添加剂最佳浓度为:1.5%海藻酸钠、4%SiO_2、0.3%CaCO_3。并对固定化颗粒密度、通透性、机械强度及菌体包埋量等性能做评价。
第三章:筛选出一株能同化乙醇的菌株,经实验室扩大培养后采用有利于微生物细胞生长的溶胶—凝胶包埋法将其制备成固定化小球。就乙醇菌的不同固定化条件进行实验比较,得出一种最佳乙醇菌固定化方法,即以10%聚乙烯醇为包埋主要载体,添加1.5%海藻酸钠防止固定化小球间粘联,加入0.01g功能化的碳纳米管来增加菌体吸附量,增强固定化小球通透性。将得到的固定化乙醇氧化菌小球与溶解氧电极组合,建立一种乙醇微生物传感体系。考察了温度、pH、盐离子浓度、菌种保存时间等对该传感体系的影响。该体系能够检测0.002%-0.2(V/V)范围内乙醇,RSD=6.3%。
第四章:筛选得到一株好氧型甲醇氧化菌,经实验室扩大培养后,采用溶胶—凝胶包埋法将其固定。固定化颗粒与溶解氧电极组合,建立一种微生物甲醇传感体系。并考察温度、pH、盐离子浓度、菌种保存时间等实验条件对微生物甲醇传感体系的影响,该体系可以检测0.002%-0.3(V/V)范围内甲醇,RSD=4.24%。
Chapter 1:Working principle,configuration,classification and characteristics of biosensor,microbial sensors were introduced.Researching development and application of microbial sensor are reviewed.Simultaneously,methods and materials of immobilization were reported.One new kind of nanometer material,carbon nanotubes and its application in biosensor was reviewed.
Chapter 2:whether carbon nanotubes are functionalized or not are inspected, by analyzing its fluorescence spectrum,infrared spectrum and so on.Sol-gel embedding method is used to prepare immobilized pellets.We compared different methods and materials which were used for embedding microbe. Affecting fators of sol-gel are carded on optimizing choice,such as, concertration of silicon dioxide(SiO_2),calcium carbonate(CaCO_3),sea alginic acid sodium and so on.The experimental results testified when 1.5% sea alginic acid sodium,4%SiO_2,0.3%CaCO_3 ale added to sol-gel, characters of immobilizing balls is the best.Permeability,mechanical strength as well as quantity of microbe embedded have been made appraisal respectively.
Chapter 3:Sol-gel embedding method has been used to immobilize microorganism cell of ethanol.Optimizing terms of immobilization are made by comparing different methods and materials.The result is that 10% polyvinyl alcohol is main materials,added 1.5%sea alginic acid sodium to sol-gel to prevent pellets from adhibitting.And 0.01g CNTs was used for increasing quantity of microbe embedded and pervasion.One kind of ethanol microbial sensing system has been constructed by combining immobilized cell with dissolved oxygen electrode.Effects of sensing system are inspected, including temperature,pH,salt ion concentration,preserving time of microbe immobilized and so on.Comcentration of ethanol from 0.002%to 0.2(V/V) can be detected by this system.RSD=6.3%.
Chapter 4:Methanol oxidation fungus is gained and expandly cultured in laboratory.Sol-gel embedding method is used for immobilizing methanol bacterias.One kind of microbial methanol sensing system is established by combining dissolved oxygen electrode with embedding methanol bacterias. Effects are inspected of this system,such as,temperature,pH,salt ion concentration,preserving time of microbe immobilized and so on. Comcentration of methanol from 0.002%to 0.3(V/V) can be detected by the system,RSD=4.24%.
第二章:通过对碳纳米管的荧光光谱、红外光谱的分析,考察碳纳米管是否被功能化。采用溶胶—凝胶包埋法,制备固定化小球,就微生物细胞不同固定化方法和载体进行实验比较。对影响溶胶—凝胶固定化的添加剂二氧化硅(SiO_2)、碳酸钙(CaCO_3)、海藻酸钠等浓度进行了考察,得出添加剂最佳浓度为:1.5%海藻酸钠、4%SiO_2、0.3%CaCO_3。并对固定化颗粒密度、通透性、机械强度及菌体包埋量等性能做评价。
第三章:筛选出一株能同化乙醇的菌株,经实验室扩大培养后采用有利于微生物细胞生长的溶胶—凝胶包埋法将其制备成固定化小球。就乙醇菌的不同固定化条件进行实验比较,得出一种最佳乙醇菌固定化方法,即以10%聚乙烯醇为包埋主要载体,添加1.5%海藻酸钠防止固定化小球间粘联,加入0.01g功能化的碳纳米管来增加菌体吸附量,增强固定化小球通透性。将得到的固定化乙醇氧化菌小球与溶解氧电极组合,建立一种乙醇微生物传感体系。考察了温度、pH、盐离子浓度、菌种保存时间等对该传感体系的影响。该体系能够检测0.002%-0.2(V/V)范围内乙醇,RSD=6.3%。
第四章:筛选得到一株好氧型甲醇氧化菌,经实验室扩大培养后,采用溶胶—凝胶包埋法将其固定。固定化颗粒与溶解氧电极组合,建立一种微生物甲醇传感体系。并考察温度、pH、盐离子浓度、菌种保存时间等实验条件对微生物甲醇传感体系的影响,该体系可以检测0.002%-0.3(V/V)范围内甲醇,RSD=4.24%。
Chapter 1:Working principle,configuration,classification and characteristics of biosensor,microbial sensors were introduced.Researching development and application of microbial sensor are reviewed.Simultaneously,methods and materials of immobilization were reported.One new kind of nanometer material,carbon nanotubes and its application in biosensor was reviewed.
Chapter 2:whether carbon nanotubes are functionalized or not are inspected, by analyzing its fluorescence spectrum,infrared spectrum and so on.Sol-gel embedding method is used to prepare immobilized pellets.We compared different methods and materials which were used for embedding microbe. Affecting fators of sol-gel are carded on optimizing choice,such as, concertration of silicon dioxide(SiO_2),calcium carbonate(CaCO_3),sea alginic acid sodium and so on.The experimental results testified when 1.5% sea alginic acid sodium,4%SiO_2,0.3%CaCO_3 ale added to sol-gel, characters of immobilizing balls is the best.Permeability,mechanical strength as well as quantity of microbe embedded have been made appraisal respectively.
Chapter 3:Sol-gel embedding method has been used to immobilize microorganism cell of ethanol.Optimizing terms of immobilization are made by comparing different methods and materials.The result is that 10% polyvinyl alcohol is main materials,added 1.5%sea alginic acid sodium to sol-gel to prevent pellets from adhibitting.And 0.01g CNTs was used for increasing quantity of microbe embedded and pervasion.One kind of ethanol microbial sensing system has been constructed by combining immobilized cell with dissolved oxygen electrode.Effects of sensing system are inspected, including temperature,pH,salt ion concentration,preserving time of microbe immobilized and so on.Comcentration of ethanol from 0.002%to 0.2(V/V) can be detected by this system.RSD=6.3%.
Chapter 4:Methanol oxidation fungus is gained and expandly cultured in laboratory.Sol-gel embedding method is used for immobilizing methanol bacterias.One kind of microbial methanol sensing system is established by combining dissolved oxygen electrode with embedding methanol bacterias. Effects are inspected of this system,such as,temperature,pH,salt ion concentration,preserving time of microbe immobilized and so on. Comcentration of methanol from 0.002%to 0.3(V/V) can be detected by the system,RSD=4.24%.
引文
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