Question

What do we mean by

Answer 1

The mixture of proteins can be separated by salting-out. Specify the reagent that is used for this purpose is (NH₄)₂SO_4.

Recovery and purification of the creatinine amidohydrolase so produced can be accomplished by conventional methods. For example, the cells are demolished by ultrasonication, mechanical disintegration with glass beads, a French press, or the action of surfactants, etc., and a crude extract is obtained.

This crude extract can be purified by various well-known methods such as salting out with ammonium sulphate or Glauber's salt, metallic coagulation with salts such as magnesium chloride or calcium chloride, coagulation with agents such as protamine or polyethylene imine, ion exchange chromatography with DEAE (diethylaminoethyl)-sepharose (Pharmacia) or CM (carboxymethyl)-cellulose (Pharmacia), etc., and gel filtration with Sephacryl S-300 (Pharmacia), etc., thereby obtaining a purified creatinine amidohydrolase.