Question

Use two enzyme to hydrolysis protein， after enzyme A hydrolysis，need to inactive Enzyme A, then add enzyme B, or can directly add enzyme B to continue hydrolysis ??

Answer 1

Enzyme B can be directly added to continue hydrolysis. When two enzymes are necessary to hydrolyze a protein then there are two conditions for using the enzymes. If the optimum activity of both enzymes occur at same temperature and pH then both enzymes can be used simultaneously. If the optimum activity of both enzymes occur at different temperature and pH then after using first enzyme, temperature and pH are adjusted for second enzyme and then second enzyme is directly added no need for inactivation of previously added enzyme. The process can be repeated if multiple enzymes are required for complete hydrolysis of protein. Inactivation of enzyme/ enzymes is required only after complete hydrolysis of protein and before centrifuge.