Dephosphorylation was kept to less than 10 % of the total phosphorylated substrate, and the reaction was linear with respect to enzyme concentration and time.

However, when enzyme concentrations are higher, these complex kinetics do not manifest.

The second assumption is that the total enzyme concentration does not change over time, thus .

In stimulating detoxification pathways (enzyme concentrations and activity).

Ligase will also work with blunt ends, although higher enzyme concentrations and different reaction conditions are required.

The secretin-induced rapid flow of water results in lower and often unreliable enzyme concentrations.

It also follows that , where is the enzyme concentration.

Thus it holds if the enzyme concentration is much less than the substrate concentration.

However, since some enzyme is always inhibited from converting the substrate to product, the effective enzyme concentration is lowered.

This is true for many enzyme-catalyzed reactions, provided that the reactant concentration is much greater than the enzyme concentration which controls the rate.