If I am using substrate concentration that is much lower than the reported Km of an enzyme, say for example 30x lower, is there a way to compensate for that? Can I just add an excess of enzyme to ensure product turn over?
Hi there,
What are you looking for? If it's just about converting a substrate into product, you actually have 2 alternatives: taking the time to reach the completion of the reaction or forcing the reaction by adding more enzyme and therefore shortening the laps of time needed to reach completion.
Your answer makes sense and I can probably add an excess of the enzyme to avoid waiting time. Thanks!
Hi Erome. Km is half of a substrate concentration required to obtain maximal enzyme activity.
There is nothing that says you cant use substrate concentrations below the Km to measure enzyme activity for your particular assay.
The Km is only a constant that says how fast an enzyme can go.
It is like a car that can go from 0-60 in 10 secs. The half time is maybe ~5 sec (~Km) to reach full speed.
You don't have to factor in the 0-60 constant when, say, you are driving to do some shopping. The same goes with enzymes.
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