What is ‘enzyme activity’
Activity is quoted in units per ml (U/ml), in other words nmol per min per ml (if unit definition B has been adopted). Thus activity values expressed in units are also subject to an illusory 1000-fold ‘increase’ if one switches from unit definition A to unit definition B. Again there cannot be any confusion if activity is expressed in terms of nmol per min per ml rather than units per ml.
Since activity relates to concentration it follows that two vials of enzyme can contain the same number of units (in total) but have different activities (concentrations).
What is specific enzyme activity?
Specific enzyme activity (usually stated simply as ‘specific activity’) is the number of enzyme units per ml divided by the concentration of protein in mg/ml. Specific activity values are therefore quoted as units/mg or nmol/min/mg (if unit definition B is applied).
Specific activity is an important measure of enzyme purity and values for different batches of a pure enzyme should be the same, within normal experimental error.
Serial dilutions of an enzyme solution will have different enzyme activity values, but identical specific activity values because in calculating specific activity the numerator (units/ml) and denominator (mg/ml) are affected equally by sample dilution.
Although specific activity is very different from activity, the calculation of specific activity nonetheless is dependent on the activity value, and therefore the stated specific activity value will also be dependent on the enzyme unit definition. Batches that are below the expected specific activity value may contain impurities or enzyme molecules that have become denatured.
Factors affecting enzyme activity
In this section we discuss why one enzyme may have different measured activity values in different labs. By this we mean real differences in measured activity, not apparent differences caused by the use of different unit definitions.
The conditions under which an assay is carried out will influence the reported activity values. For example, assays typically are carried out at a temperature between 20-37oC. Generally speaking, an enzyme will be more active at 37oC than at 20oC.
The definition of the enzyme unit would be better expressed thus:
1 unit (U) is the amount if enzyme that catalyses the reaction of 1 nmol of substrate per minute under standard conditions.