I'm writing a report on enzyme activity and characterisation and i need help with understanding this: we were told to report on our results for a 1/10 diluted acid phosphatase. I want to know why we need to dilute the enzyme in the first place and why not use the crude enzyme lysate?
The rate of an enzyme-catalyzed reaction is directly proportional to the enzyme concentration. If the enzyme is too concentrated, the reaction may go too fast for the assay. For example, it might use up all the substrate, or the amount of product formed may be too high for the measurement technique. In such a situation, you might not be able to tell the difference between different levels of enzyme activity.
If you are trying to measure the initial rate of the reaction for the sake of studying the kinetics, it is important to keep the rate of product formation within a certain range at which the product concentration increases in direct proportion to the reaction time (i.e. the initial rate), so adjusting the enzyme concentration accordingly is necessary.
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