Whole-cell amperometric biosensor for screening of cytochrome P450 inhibitors

Tal Yoetz-Kopelman, Carmit Porat-Ophir, Yosi Shacham-Diamand, Amihay Freeman

Research output: Contribution to journalArticlepeer-review

Abstract

A fast and cost-effective whole-cell electrochemical biosensor aiming at early-stage screening of potential inhibitors of cytochrome P450 is proposed and its feasibility demonstrated. Sensing is performed by monitoring the decrease in the electrochemical signal generated by the oxidation of the product of the enzymatic reaction of cytochrome P450 BM3 expressed in E. coli cells. The system is self-maintained and does not require enzyme purification or external addition of NADPH or other cofactors. Measurements were performed simultaneously at eight chips at low positive potential of 100 mV vs. Ag/AgCl under continuous stirring. Kinetic analysis of aniline determination by the whole-cell system was performed and the concentration of aniline for the inhibition studies was selected accordingly. Three known inhibitors - imidazole, metyrapone and 1-aminobenzotriazole (ABT) - were tested and their inhibition profiles characterized. Imidazole was found to be the most potent inhibitor of the three. To the best of our knowledge, the system developed enables for the first time rapid and cheap cytochrome P450 inhibitors detection by a disposable whole-cell electrochemical chip, combined with the advantages of a whole-cell system, without neither enzyme purification nor NADPH addition. Furthermore, the system developed also provides capability of performing aniline detection e.g. for environmental monitoring, by means of electrochemical biosensing.

Original languageEnglish
Pages (from-to)392-399
Number of pages8
JournalSensors and Actuators, B: Chemical
Volume223
DOIs
StatePublished - 1 Feb 2016

Keywords

  • Aniline
  • Cytochrome P450 BM3
  • E. coli
  • Electrochemical biosensor
  • Inhibitors screening
  • Whole-cell biosensor

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