enzyme inhibition experiments?

Question

experiment done in my lab taking constant voluumes of a unknown substrate,elastase enzyme and increasing concentrations of elastase enzyme inhibitor. A graph between absorbency and enzyme inhibitor concentration revealed the absorbency increased as we increased the enzyme inhibitor concentration. Can i say from the experiment what is the type of inhibition. If not what experiment should be done to find the mode of inhibition of enzyme

Answer 1

As happyman said you need to do the Lineweaver-Burk plot.
But that is 1/V vs 1/[S], where V is the rate of the reaction which you calculate with the help of the measured absorbance and [S] the concentration of substrate. So you need to find the V for different substrate concentrations.
First you need to do the plot in the abscence of any inhibitor, calculate Vmax and Km, then repeat the experiment in the presence of a moderate concentration of inhibitor and determine the "new" values for Vmax and Km. Depending on their change you can classify your inhibitor.

Thus the experiment you did can only give you a hint approximately which concentration of inhibitor to use when determining V vs [S] in the presence of inhibitor, to find the new Vmax and Km.

Answer 2

Your not looking at the absorbancy but at the rate of consumption of substrate or the rate of formation of product. And you are not changing the enzyme concentration but the substrate concentration.
You use a Lineweaver-Burke plot to determine the type of enzyme. And note whether Km or Vmax is changing.
If your enzyme follows Michaelis-Menton kinetics (the simplest with only one active site for one substrate) you will be able to know whether its a non-competitive, uncompetitive or competitive enzyme.

Answer 3

dont know what it even is