Insecticide detection through protein engineering of Nippostrongylus brasiliensis acetylcholinesterase B

Holger Schulze, Susanne B Muench, Francois Villatte, Rolf D Schmid, Till T Bachmann

Research output: Contribution to journalArticlepeer-review

Abstract

The sensitivity of acetylcholinesterase (AChE) biosensors for insecticide detection could be increased substantially by engineering AChE B of Nippostrongylus brasiliensis. The introduction of 10 single and 4 double mutations into the AChE peptide chain led to an increase in sensitivity to 10 of the 11 insecticides tested. The combination of three mutants with the wild-type enzyme in a multienzyme biosensor array enabled the detection of 11 out of the 14 most important organophosphates and carbamates at concentrations below 10 microg/kg, the maximum residue limit of infant food. The detection limit for pirimiphos methyl could be reduced from 10 microg/L to a value as low as 1 ng/L (3.5 x 10(-)(12) mol/L). The newly created biosensors exhibited an extraordinary high storage stability. There was no loss of sensitivity of N. brasiliensis AChE B, immobilized on screen-printed, disposable electrodes, even after 17-month storage at room temperature.
Original languageEnglish
Pages (from-to)5823-30
Number of pages8
JournalAnalytical Chemistry
Volume77
Issue number18
DOIs
Publication statusPublished - 2005

Keywords

  • Acetylcholinesterase
  • Animals
  • Biocompatible Materials
  • Biosensing Techniques
  • Catalysis
  • Cholinesterase Inhibitors
  • Gene Expression
  • Insecticides
  • Models, Molecular
  • Mutation
  • Nippostrongylus
  • Oxidation-Reduction
  • Protein Engineering
  • Protein Structure, Tertiary
  • Sensitivity and Specificity
  • Temperature

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