Principles of Basic and Advanced Techniques Used for the Purification of Enzymes of Industrial Importance

Recovery and purification are very critical and essential steps in the downstream processing (DSP) of industrially important enzymes and compounds. Purification process involves the enrichment of enzyme proportion by several hundred folds by employing various biophysical–biochemical tools and techniques. The purity and yield depend on the efficacy of suitable methods used in isolation, purification, and packaging of the end product. In the current topic, we have discussed the basic principles and workflow of enzymes isolation and their purification that involves various methods that includes cell disruption, centrifugation, filtration, and precipitation (by inorganic or organic solvents or isoelectric pH) in combination with various types of chromatographic techniques. It is very essential to verify the purity and activity of enzyme at various stages of purification steps by PAGE gel analysis and total activity and specific activity of enzymes. The downstream processing for some of the industrially important enzymes such as amylase, glucose isomerase, lipase, and pharmaceutically important enzymes have been discussed. Thus, understanding the basic principles of enzyme purification will definitely help in designing strategies that can influence the overall economy of enzyme manufacturing industry and eases the handling process.