Answer
Salting out is the process by which proteins can be precipitated from the solution based on diferrence in salt concentration. The mechanism as followes: Generally proteins are of two types based on water interaction i.e., hydrophobic and hydrophilic, in aqueous (salt- ionic strngth) solution hydrophobic aminoacids got protected where as hydrophilic molecules are interact with molecules of solvation and forms hudrogen bond with adjacent water molecules, when this salt solution increased the metal ions can attract the surrounding molecules as a result less availability of water molecules to interact with the charged part of the protein which can leads to demand for solvent molecules and the protein-protein interactions are stronger than solvent-solute interactions and the protein molecules precipitated by forming hydrophobic interactions with each other. The salt is used is generally ammonium sulphate and its dosage is increased step by step and each protein precipitated is dissolved in various buffers and tested for total protein content and desired protein. The precipitated protein is removed by centrifugation followed by increasing the concentration of ammonium sulphate solution can seperates the next desired protein which we require.
Work Step by Step
Salting out is the process by which proteins can be precipitated from the solution based on diferrence in salt concentration. The mechanism as followes: Generally proteins are of two types based on water interaction i.e., hydrophobic and hydrophilic, in aqueous (salt- ionic strngth) solution hydrophobic aminoacids got protected where as hydrophilic molecules are interact with molecules of solvation and forms hudrogen bond with adjacent water molecules, when this salt solution increased the metal ions can attract the surrounding molecules as a result less availability of water molecules to interact with the charged part of the protein which can leads to demand for solvent molecules and the protein-protein interactions are stronger than solvent-solute interactions and the protein molecules precipitated by forming hydrophobic interactions with each other. The salt is used is generally ammonium sulphate and its dosage is increased step by step and each protein precipitated is dissolved in various buffers and tested for total protein content and desired protein. The precipitated protein is removed by centrifugation followed by increasing the concentration of ammonium sulphate solution can seperates the next desired protein which we require.
