Unit Operations of Enzyme Purification

Abstract

The successful purification of an enzyme from a bacterial or tissue source usually follows a general separation flowsheet consisting of the following general processing steps: (1) Starting material preparation and cell harvesting, which includes tissue removal, grinding, cellular disruption and centrifugation to produce a crude extract; (2) Crude extract separation, which includes DEAE, or other charged support, either fractional or total product precipitation, or product separation by size, gel exclusion, or membrane sizing; (3) Secondary separation of the partially purified product, which usually is a combination of the above separation steps; (4) Pre- and post-treatment of the product (e.g. salt removal, water removal, and pH adjustment) to prepare for the next separation operation, which is frequently time-consuming and results in a possibility of product loss.