通过将多酚氧化酶固定在导电共聚物中来制备生物传感器及其在测定红酒中酚类化合物的应用。
Preparation of biosensors by immobilization of polyphenol oxidase in conducting copolymers and their use in determination of phenolic compounds in red wine.
作者信息
Böyükbayram A Elif, Kiralp Senem, Toppare Levent, Yağci Yusuf
机构信息
Department of Chemistry, Middle East Technical University, 06531 Ankara, Turkey.
出版信息
Bioelectrochemistry. 2006 Oct;69(2):164-71. doi: 10.1016/j.bioelechem.2006.01.006. Epub 2006 Feb 23.
Electrochemically produced graft copolymers of thiophene capped polytetrahydofuran (TPTHF1 and TPTHF2) and pyrrole were achieved by constant potential electrolysis using sodium dodecylsulfate (SDS) as the supporting electrolyte. Characterizations were based on Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). Electrical conductivities were measured by the four-probe technique. Novel biosensors for phenolic compounds were constructed by immobilizing polyphenol oxidase (PPO) into conducting copolymers prepared by electropolymerization of pyrrole with thiophene capped polytetrahydrofuran. Kinetic parameters, maximum reaction rate (V(max)) and Michaelis-Menten constant (K(m)) and optimum conditions regarding temperature and pH were determined for the immobilized enzyme. Operational stability and shelf-life of the enzyme electrodes were investigated. Enzyme electrodes of polyphenol oxidase were used to determine the amount of phenolic compounds in two brands of Turkish red wines and found very useful owing to their high kinetic parameters and wide pH working range.
以十二烷基硫酸钠（SDS）作为支持电解质，通过恒电位电解法制备了噻吩封端聚四氢呋喃（TPTHF1和TPTHF2）与吡咯的电化学接枝共聚物。表征基于傅里叶变换红外光谱（FTIR）和扫描电子显微镜（SEM）。通过四探针技术测量电导率。通过将多酚氧化酶（PPO）固定在由吡咯与噻吩封端聚四氢呋喃电聚合制备的导电共聚物中，构建了用于酚类化合物的新型生物传感器。测定了固定化酶的动力学参数、最大反应速率（V(max)）和米氏常数（K(m)）以及关于温度和pH的最佳条件。研究了酶电极的操作稳定性和保质期。多酚氧化酶的酶电极用于测定两个品牌土耳其红葡萄酒中酚类化合物的含量，由于其高动力学参数和宽pH工作范围，发现非常有用。
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