聚邻苯二胺/辣根过氧化物酶电极上的生物电催化研究

学位：理学硕士院系专业：化学化工学院化学系_物理化学(含化学物理)学号：1992440

Bioelectrocatalytic Oxidation of p Hydroxyphenol at Horseradish Peroxidase/Poly o phenylenediamine Membrane Electrode: A Kinetic Study

用酶的电化学固定化方法制备辣根过氧化物酶/聚邻苯二胺膜（HrP/PPd）电极.以对苯二酚和邻苯二胺在该酶电极上的氧化为模型体系，讨论了有机物生物催化氧化的一般动力学模式以及影响酶电极检测的因素.邻苯二胺可在辣根过氧化物酶的存在下发生聚合，因而它在HrP/PPd电极上的氧化过程比对苯二酚的复杂.测定了不同电位下对苯二酚在HrP/PPd电极上的反应动力学参数，并根据实验结果确定该有机物检测的合适工作电位.A horseradish peroxidase (HRP)/poly o phenylenediamine(PPD) membrane electrode was prepared by electrochemical immobilization of enzyme. The oxidations of p hydroxyphenol and o phenylenediamine as model systems were studied at the enzyme electrode, thereby the general kinetic scheme of bioelectrocatalytic oxidation of organic compounds and the Factors inFluencing on the detection with the HRP/PPD electrode were discussed. o Phenylenediamine was Found to be polymerized in the presence of HRP, hence its oxidation process at the HRP/PPD electrode is more complicated than that of p hydroxyphenol.The kinetic parameters of p hydroxyphenol oxidation at the enzyme electrode were measured at diFFerent potentials, and the optimum operational potential For p hydroxyphenol detection was determined.国家自然科学基金;国家教委博士点科研基

Electrochemical Immobilization of HRP and Properties of HRP/ P OPD Enzyme Membrane Electrodes

利用电化学固定化方法制备含辣根过氧化物酶的聚邻苯二胺（HrP/P-OPd）膜电极，改变聚合用溶液的酸度和所含支持电解质的成分，研究酶的固定化过程及其对所得酶电极性能的影响.结果表明依电聚合条件而异，酶可能以不同途径进入聚合物基质中.支持电解质的品种会影响酶膜的形成速度、组成和稳定性.所得HrP/P-OPd膜电极可在溶液不含电子传递体的条件下催化H2O2的还原，电聚合过程中进入酶膜的寡聚体可能充当酶的电子传递体By the electrochemical immobilization technique , horseradish peroxidase was entrapped into poly o phenylenediamine to prepare the enzyme membrane electrodes. The mechanism of enzyme immobilization was studied with changing the acidity of polymerization solution and the kinds of supporting electrolytes, and the properties of the resulting enzyme electrodes were examined. The results show that,depending on the immobilization conditions, the mechanism of entrapment of HRP into the polymer matrix is diFFerent.The supporting electrolytes have great inFluences on the growth rate, composition and stability of the enzyme membrane. The resulting enzyme electrode exhibits the activity to catalyze the electroreduction of hydrogen peroxide in the absence of any electron transFer mediator in solution. It was illustrated that the oliogomers ,which is produced in the course of o phenylenediamine polymerization and then incorporated in the enzyme membrane, may play a role of electron transFer mediator.国家自然科学基金;国家教委博士点基

Electron TransFer of Horseradish Peroxidase Entrapped in Poly-o-phenylenediamine Membrane Electrode

利用酶的电化学固定化方法制备合辣根过氧化物酶的聚邻苯二胺膜电极，研究其伏安行为及对H2O2还原的生物电催化作用.结果表明，在所述生物电催化反应中酶与聚合物基质发生了直接电子传递.但对新制的酶电极而言，电聚合时生成并包理在酶膜中的寡聚体可作为电子传递体加速氧化态酶的再生.根据酶电极电流响应实验曲线的拟合，发现氧化态酶的再生速度随电极电位的变化表观上符合TAfEl关系式.提出了酶反应动力学参数的测定方法.The poly-o-phenylenediamine membrane electrode containing horseradish peroxi-dase was prepared by means of the electrochemical immobilization of enzyme, and thevoltammetric behavior of the resulting enzyme electrode and its bioelectrocatalysis to hydro-gen peroxide reduction was studied.It was Found that the direct electron tansFer between theenzyme and the polymer matrix takes place in the biocatalytic processes.For the newly pre-pared enzyme electrode, however, the oligomers, which are Formed during the polymeriza-tion of o-phenylenediamine and then incorporated in the enzyme membrane, can play a role ofelectron transFer mediators to accelerate the regeneration of the oxidized enzyme.From theFitting of experimental current response curves, it can be Found that the change in the regen-eration rates of the oxidized enzyme with the electrode potentials obeys apparantly the TaFelrelationship.The procedure in the measurement of kinetic parameters For enzyme reactionswas suggested.国家自然科学基金;国家教育委员会博士点基

Electron transfer of horseradish peroxidase entrapped in poly-o-phenylenediamine membrane electrode

The poly-o-phenylenediamine membrane electrode containing horseradish peroxidase was prepared by means of the electrochemical immobilization of enzyme, and the voltammetric behavior of the resulting enzyme electrode and its bioelectrocatalysis to hydrogen peroxide reduction was studied. It was found that the direct electron tansfer between the enzyme and the polymer matrix takes place in the biocatalytic processes, For the newly prepared enzyme electrode, however, the oligomers, which are formed during the polymerization of o-phenylenediamine and then incorporated in the enzyme membrane, can play a role of electron transfer mediators to accelerate the regeneration of the oxidized enzyme, From the fitting of experimental current response curves, it can be found that the change in the regeneration rates of the oxidized enzyme with the electrode potentials obeys apparantly the Tafel relationship. The procedure in the measurement of kinetic parameters for enzyme reactions was suggested

艾滋病毒重组抗原及第三代艾滋病毒抗体EIA诊断试剂盒的研制

近年来中国的艾滋病毒(HIV)感染人数以每年30％的速度增长,HIV感染者总数已超过60万人,感染者已从吸毒人员、献血员等高危人群扩展到社会各个阶层,我国已处于艾滋病泛滥前的关键时期。HIV诊断试剂盒是HIV防制工作中最重要的技术工具。用于人群筛检的HIV诊断试剂盒已历经三代,第一代试剂盒主要使用来自细胞培养的HIV病毒的裂解产物作为抗原;第二代产品使用基因工程重组抗原和/或人工合成肽作为抗原,使包被抗原中免疫优势区的比例明显增强,从而提高了试剂的灵敏度,而且标准化生产的相对更纯的抗原的使用使试剂的特异性和重复性均大为提高。前两代试剂均采用间接法原理,即先以抗原包被聚丙乙烯板,再加入待检血清,最后加入酶标记的抗人Ig