Insecticide Detection through Protein Engineering of Nippostrongylus brasiliensis Acetylcholinesterase B

详细信息    查看全文

• 作者：Holger Schulze ; Susanne B. Muench ; Francois Villatte ; Rolf D. Schmid ; and Till T. Bachmann
• 刊名：Analytical Chemistry
• 出版年：2005
• 出版时间：September 15, 2005
• 年：2005
• 卷：77
• 期：18
• 页码：5823 - 5830
• 全文大小：521K
• 年卷期：v.77,no.18(September 15, 2005)
• ISSN：1520-6882

文摘

The sensitivity of acetylcholinesterase (AChE) biosensorsfor insecticide detection could be increased substantiallyby engineering AChE B of Nippostrongylus brasiliensis.The introduction of 10 single and 4 double mutations intothe AChE peptide chain led to an increase in sensitivityto 10 of the 11 insecticides tested. The combination ofthree mutants with the wild-type enzyme in a multienzymebiosensor array enabled the detection of 11 out of the 14most important organophosphates and carbamates atconcentrations below 10 g/kg, the maximum residuelimit of infant food. The detection limit for pirimiphosmethyl could be reduced from 10 g/L to a value as lowas 1 ng/L (3.5 × 10^-12 mol/L). The newly createdbiosensors exhibited an extraordinary high storage stability. There was no loss of sensitivity of N. brasiliensisAChE B, immobilized on screen-printed, disposable electrodes, even after 17-month storage at room temperature.