My approach is to increase the coupling enzyme concentration until there is no further increase in the rate of the complete reaction at saturating substrate concentrations, then double it, at least. This assures that the activity of the coupling enzyme will not be rate limiting.

My approach is to increase the coupling enzyme concentration until there is no further increase in the rate of the complete reaction at saturating substrate concentrations, then double it, at least. This assures that the activity of the coupling enzyme will not be rate limiting.

Dear Yacoba, thank you for your request, with a hope that you find in these links, a useful references to find the correct answer on your question..

"Enzymes are proteins functioning as catalysts that speed up reactions by lowering the activation energy. A simple and succinct definition of an enzyme is that it is a biological catalyst that accelerates a chemical reaction without altering its equilibrium." : https://www.eolss.net/Sample-Chapters/C03/E6-54-02.pdf

2.3 Enzyme Kinetics

Coupled Enzyme System: https://iubmb.onlinelibrary.wiley.com/doi/pdf/10.1016/0307-4412%2890%2990025-J

Just try to always remembering - my dear- that enzymes are "organic" and "biological" catalysts: https://www.cliffsnotes.com/study-guides/biology/biochemistry-i/enzymes/enzymes-are-catalysts

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following

Thank you Adam and Aisha.

Adam, that is what I did . however the turnover number I calculate from the kinetic reaction is similar to that of just the coupled enzyme. That has me a little confused.

You should see no catalytic turnover when the primary enzyme is omitted. If you are seeing catalysis under those circumstances, you need to reconsider the enzyme you chose as the coupling enzyme.