Electro-catalysis by immobilised human flavin-containing monooxygenase isoform 3 (hFMO3).

Castrignanò, Silvia; Sadeghi, Sheila J.; Gilardi, Gianfranco

Human flavin-containing monooxygenases are the second most important class of drug-metabolizing enzymes after cytochromes P450. Here we report a simple but functional and stable enzyme-electrode system based on a glassy carbon (GC) electrode with human flavin-containing monooxygenase isoform 3 (hFMO3) entrapped in a gel cross-linked with bovine serum albumin (BSA) by glutaraldehyde. The enzymatic electrochemical responsiveness is characterised by using well-known substrates: trimethylamine (TMA), ammonia (NH), triethylamine (TEA), and benzydamine (BZD). The apparent Michaelis–Menten constant ( K′) and apparent maximum current ( I′) are calculated by fitting the current signal to the Michaelis–Menten equation for each substrate. The enzyme-electrode has good characteristics: the calculated sensitivity was 40.9 ± 0.5 mA mol L cm for TMA, 43.3 ± 0.1 mA mol L cm for NH, 45.2 ± 2.2 mA mol L cm for TEA, and 39.3 ± 0.6 mA mol L cm for BZD. The stability was constant for 3 days and the inter-electrode reproducibility was 12.5%. This is a novel electrochemical tool that can be used to investigate new potential drugs against the catalytic activity of hFMO3.

MONOOXYGENASES; OXYGENASES; FLAVINS; COENZYMES; CYTOCHROMES

Analytical & Bioanalytical Chemistry, 2010, Vol 398, Issue 3, p1403

1618-2642

Academic Journal

10.1007/s00216-010-4014-z