 Phenazine methosulfate

Phenazine methosulfate is a salt containing a methylphenazinium cation and a methyl sulfate counterion. Its overall formula is ${\mathrm{C}}_{14}{\mathrm{H}}_{14}{\mathrm{N}}_2{\mathrm{O}}_4\mathrm{S}$. The redox-active phenazinium system makes the compound useful as an artificial electron carrier in coupled biochemical measurements.^[ref]

The original methanol dehydrogenase work used phenazine methosulfate between the enzyme reaction and an optical readout involving DCPIP. A reducing equivalent transferred from substrate through the carrier can reduce the dye, so a change in absorbance reports the coupled system. The investigators used a spectrophotometer with 10-mm optical paths and calibrated the dye's absorption at 600 nm.^[ref]

An original measurement example separates concentration from mechanism. Suppose a dye's initial absorbance is 0.80 and its corrected final absorbance is 0.40. If the Beer-Lambert law applies with unchanged optical path length and coefficient, the absorbing dye concentration has halved. That observation alone does not determine the concentration of the enzyme or prove which substrate supplied the electrons.

The carrier and reporter are separate experimental components. Controls without enzyme or without substrate help distinguish the intended coupled reaction from background reduction. A carrier compatible with one enzyme system is not automatically the natural physiological electron acceptor of that enzyme.