I am trying to re-determine the Km of the enzyme I'm working with. For some reasons, I'm trying to do this using a coupled assay. I am having trouble with that, because the Km that I am finding is more than two times what it should be. I was told that it could be because of a reduced activity of the second enzyme in this coupled assay.

I am having a hard time understanding this statement. Can someone explain to me why a reduced activity of the second enzyme would give me a greater Km of a substrate of the first enzyme?

Similar questions and discussions