Question

why do phosphotase experiments i.e.

experiments ( biochemistry lab) with enzyme phosphatase require different pH buffers with different concentrations ( volume )of buffers ?

Answer 1

In any enzymatic experiments, the activity of the enzyme plays a major role in the catalysis. The major parameter for most enzymes is that their pH can influences the active site directly via ionizing the different amino acids present in their active site and changes the activity and reaction rate. pH also affects the tertiary structure of the enzyme. Since the activity od enzymes are very sensitive to pH it is necessary to use different pH concentrations to access for the optimal pH conditions at which the enzymes can work most efficiently. Again there are different types of phosphatases present that differ in their pH optima. So using a wide range of pH in the buffer will help us to determine the optima and the highest enzyme activity of phosphatases. The buffer solution is also capable of influencing the buffer capacity, i.e the greater the concentration, the greater the buffering. The of the buffer solution is inversely proportional to the concentration of the buffer and it is because of this relationship, that most experiments use wide range of buffer concentration equivalent to the pH for experiments with enzymes.