I need to calculate the residual activity after I have gotten the Absorbance @ 480 nm wavelength at interval of 30 seconds. But I dont want to used protein concentration as in the formula below: Enzyme activity = (change in OD@480)/(Vol of supernatent used x protein conc).
I dont have the protein conc.
You could try to quantify the amount of H2O2 consumed per unit time. H2O2 has absorbance at around 240nm. You need to titrate against KMnO4 to determine exact concentration of peroxide and then generate a calibration curve of different concentrations versus O.D. From this curve you can quantify the amount of peroxide being consumed by the enzyme per unit time by simply measuring the O.D of H2O2. However to get specific activity of your enzyme you still need to know the protein concentration.
If you keep the enzyme concentration always the same, then you can calculate the residual activity without dividing by the enzyme concentration.
use enzyme activity from 100% then measured residual activity.
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