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. 2008 Sep 28:(36):4354-6.
doi: 10.1039/b805447a. Epub 2008 Jul 18.

Electrochemical biosensor featuring a two-enzyme pathway and DNA for screening toxic reactive metabolites of arylamines

Minjeong So  1 , John B Schenkman, James F Rusling
Affiliations

Electrochemical biosensor featuring a two-enzyme pathway and DNA for screening toxic reactive metabolites of arylamines

Minjeong So et al. Chem Commun (Camb). .

Abstract

We demonstrate for the first time a biosensor featuring a sequential two-enzyme pathway suitable to screen potentially toxic reactive metabolites generated during metabolism.

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Figures

Fig. 1
Fig. 1
(a) SWVs for sensors featuring PDDA/DNA/(CYP1A2/DNA)2 films (denoted CYP1A2/DNA) in pH 5.5 buffer containing 50 μM Ru(bpy)3 2+ before (0 min) and after (3 min) incubations at 37 °C with 50 μM PhIP and 1 mM H2O2 (for enzyme activation) at pH 7.5. (SWV ampl. 25 mV; freq. 15 Hz; 4 mV step). (b) Influence of incubation time with 50 μM PhIP and 1 mM H2O2 on CYP1A2/DNA sensor final/initial peak ratio (●) (error bars are sd); and on PDDA/DNA/(Mb/DNA)2 control (denoted Mb/DNA) (○) (relative sd in controls from 3–5 trials: ±8%).
Fig. 2
Fig. 2
(a) SWVs of DNA/RuPVP/DNA/(CYP1A2/DNA)2/(NAT/DNA)2 2-enzyme sensors in pH 5.5 buffer before (0 min) and after (3 min) incubations at 37 °C in pH 7.5 buffer containing 50 μM PhIP, 1 mM H2O2, 0.5 mM AcCoA, 1 mM DTT and 1 mM EDTA (SWV ampl. 25 mV; freq. 15 Hz; step 4 mV). (b) Influence of enzyme incubation time on final/initial peak ratio for DNA/RuPVP/DNA/(CYP1A2/DNA)2/(NAT/DNA)2 2-enzyme sensors (●), control 1 (○) DNA/RuPVP/DNA/(CYP1A2/DNA)2 sensor with 50 μM PhIP and 1 mM H2O2, and control 2 (△) DNA/RuPVP/DNA/(NAT/DNA)2 sensor with 50 μM PhIP, 0.5 mM AcCoA, 1 mM DTT, 1 mM EDTA (relative sd in controls from 3–5 trials was ±8%).
Scheme 1
Scheme 1
Metabolism of PhIP in the two-enzyme sequence.
See this image and copyright information in PMC

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