Partitioning Techniques for Protease Purification: Exploring the Potential of Two-Phase and Three-Phase Systems

Purification of proteases is an important step in obtaining high-purity enzymes for industrial and research applications. One promising technique to achieve this goal is phase separation, which utilizes differences in molecular affinity to two or more separate phases. This chapter looks at how well two-phase and three-phase systems work for purifying proteases. It focuses on the different ways they separate, how to choose the right solvent, and the pros and cons of each system. Two-phase solutions, such as polymer-based systems and organic solvents, have been shown to work well for separating proteases based on their hydrophobic properties. Three-phase techniques also have the extra benefit of splitting the mixture into three parts, which makes the purification process more efficient and allows for more exact enzyme separation. We look at a lot of experimental studies that use two-phase and three-phase systems to see how well these methods work for purifying proteases from both native and recombinant sources. The results of these studies show that two-phase and three-phase systems could be very useful in industry, but there are problems with stability and operational costs that need to be worked out before they can be used. For the future, this chapter gives useful information that will help make protease purification procedures more efficient and less expensive.