your statement is kind of vague.
if all you are asking is how to set up an experiment with varying enzyme concentrations... well, not to be snotty but you simply vary the concentrations and record whatever activity you are looking for at each concentration.
I cannot tell you the reasonable concentrations to begin with. But lets say you started at 0 microliters and then ran the same testing with 1 microliter, then 2, then 3... or maybe 0, 10, 20, 30... you get the point. I do not work with enzymes, so I cannot tell you what a reasonable amount to use is, and it may vary depending on the source/concentration. I know the only time I had to use enzymes in my undergrad class we used something like 5 microliters of one enzyme and 30 microliters of another... this wasn't a concentration study like you described, but we could've done one like it. Use 0, 1, 2, 3, 4, 5 microliters of the one and see how the rate of reaction changes in each case.
the idea is to follow the enzyme activity over different concentrations. Better if you can follow the activity by some quantitative measurement so you can better chart the differences. How you measure this depends on the enzyme and what action it catalyzes. Different products can be measured in different ways. Often you'll use something like a spectrophotometer to measure changes versus the 0 standard.
depending on if the enzyme causes a change that is easy to see visually, you could also perhaps avoid a quantitative analysis and simply describe the different results (ie: 0 = no cloudiness, 10 = some cloudiness, 20 = more cloudiness... or whatever you could physically observe)
you construct something like this
enzyme concentration... activity
0... (whatever is observed)
10... (whatever is observed)
20... (whatever is observed)
et cetera
you might expect to see generally increased enzyme activity with increasing enzyme concentration. At some point they'll be so much enzyme and so little substrate that the rate won't increase anymore... you'll just have "excess" enzyme present with nothing to do since the substrate level will be so small in comparison.
this should be a simple example that gets you a head start.
|