Temperature
Reaction rate increases with temperature. Most enzymes show their optimal activity between 30 °C and 50 °C. Above 60 °C, enzyme denaturation occurs. The incubation temperatures in test methods
must be strictly followed.
Time
The substrate concentration falls with time as the product concentration increases. Timing is therefore important because substrate and product concentrations are changing all the time and it is one of
these that is being measured. An accurate timer must be used to measure the incubation time of an enzyme and its substrate.
pH
Any increase or decrease in pH away from the optimum will cause a decrease in enzyme activity. A marked change in pH can lead to the denaturation of an enzyme. The pH of buffers and substrates used in enzyme tests must therefore be correct.
Light rays
Ultraviolet light tends to inhibit enzyme activity while blue or red light tends to increase it. Samples for enzyme analysis must therefore be protected from direct light.
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