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基于丝网印刷碳电极和循环伏安法的甲基对硫磷微生物生物传感器检测。

Microbial biosensor for detection of methyl parathion using screen printed carbon electrode and cyclic voltammetry.

机构信息

Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre Trombay, Mumbai 400085, India.

出版信息

Biosens Bioelectron. 2011 Jul 15;26(11):4289-93. doi: 10.1016/j.bios.2011.04.027. Epub 2011 Apr 22.

Abstract

Whole cells of recombinant Escherichia coli were immobilized on the screen printed carbon electrode (SPCE) using glutaraldehyde. Recombinant E. coli was having high periplasmic expression of organophosphorus hydrolase enzyme, which hydrolyzes the methyl parathion into two products, p-nitrophenol and dimethyl thiophosphoric acid. Cells immobilized SPCE was studied under SEM. Cells immobilized SPCE was associated with cyclic voltammetry and cyclic voltammograms were recorded before and after hydrolysis of methyl parathion. Detection was calibrated based on the relationship between the changes in the current observed at +0.1 V potential, because of redox behavior of the hydrolyzed product p-nitrophenol. As concentration of methyl parathion was increased the oxidation current also increased. Only 20 μl volume of the sample was required for analysis. Detection range of biosensor was calibrated between 2 and 80 μM of methyl parathion from the linear range of calibration plot. A single immobilized SPCE was reused for 32 reactions with retention of 80% of its initial enzyme activity.

摘要

采用戊二醛将重组大肠杆菌的全细胞固定在丝网印刷碳电极(SPCE)上。重组大肠杆菌具有高周质表达的有机磷水解酶,可将甲基对硫磷水解成两种产物,对硝基苯酚和二甲硫代磷酸。对细胞固定化 SPCE 进行了扫描电子显微镜(SEM)研究。对细胞固定化 SPCE 进行了循环伏安法研究,并在水解甲基对硫磷前后记录了循环伏安图。检测是根据在+0.1 V 电位下观察到的电流变化与水解产物对硝基苯酚的氧化还原行为之间的关系进行校准的。随着甲基对硫磷浓度的增加,氧化电流也随之增加。分析仅需要 20 μl 的样品体积。从校准曲线的线性范围来看,生物传感器的检测范围在 2 到 80 μM 的甲基对硫磷之间进行校准。单个固定化 SPCE 在保留 80%初始酶活性的情况下可重复使用 32 次反应。

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