Question

Hello. Question:

I have absorption data collected from 5 samples of the same enzyme at different concentration. The data is collected over the interval of 3 minutes. How can I convert this into catalytic velocity?

Answer 1

Firstly, as we know, Enzyme Activity is a measure of the amount of active enzymes present in the given sample; which in chemical terms can be said as the no. of substrates that are converted per unit time is referred to as Enzyme Activity.

and, the quantity of light absorbed by the substance(sample taken) dissolved in a fully transmitting solvent is directly proportional to the concentration of the sample taken and the path length of the light through the solution; this is called as BEER-LAMBERT'S LAW!

according to this law, we can say that concentration and absorbance are directly proportional to each other.

A = epsilon . l . c ( product of epsilon, l , C )

where ; A = absorbance

epsilon = greek constant

l = length of the light traveled in the sample taken (solution)

c = concentration of the sample taken (solution)

first, express the velocity as the variation of absorbance per unit time, and plot a graph as a function of time and this is the absorbance graph of the solution (enzyme sample taken ) and consider the peaks for which concentration you want to calculate the absorbance values

Using the molar coefficient of the absorbing molecules in Lambert's equation; Normalize the quantity by the molar concentration of the enzyme; so as to get the catalytic velocity! ( in dimensions of time inverse ) .

thank you !!!!!