Purification of Enzymes, Dialysis, Chromatography, Electrophoresis, Molecules, Dialafiltration

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Biocatalysis and Enzyme Biotechnology Biocatalysis and Enzyme Biotechnology - Introduction Some Research Activities Involving Enzymatic Synthesis Properties of Enzymes Parameters Affecting Enzymatic Reactions Definitions of Some Parameters used for Describing Key Properties of Enzymes Medium for Enzymes Aqueous vs Organic Solvent Selectivity of Enzymes Types of Enzymes Biomimetic Catalysts Enzymes Commonly Used in Organic Synthesis Extremozymes Cofactor Dependent Enzymes and Cofactor Regeneration Modular Enzymes Coenzymes Use of Enzymes Medical and Clinical Uses Drugs for Digestive Disorders Industrial Uses Uses of Industrial Uses Textile Industry Uses of Leather Industry Uses of Paper Industry Cellulase Free Xylanases Uses of Food Industry Isolation and Purification of Enzymes

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Purification of Enzymes
Enzyme purification involves three steps, i.e., dialysis, chromatography and electrophoresis. These three techniques and their variants are briefly described in the following text.

Dialysis
Dialysis is the process that is used to remove small molecules from crude enzyme. For this, enzyme precipitate obtained in previous step is dissolved in a small quantity of buffer solution in which the enzyme was originally extracted. The solution is taken in a dialysis bag (may be a cellophane tube) and after sealing securely, the bag is suspended in either distilled water or a buffer of known molarity and ionic composition. Some other salts or chemicals may have to be added sometimes in the outer solution, to prevent the loss of enzyme activity during dialysis.

The dialysis is carried out for a few hours with regular change of the outer solution or distilled water. At large scale enzyme purification dialafiltration instead of dialysis is used. The enzyme solution is filtered for small molecules through a membrane generally mounted on a fibrous support, by pressure driven operation.

An Arrangement for Dialysis to Purify an Enzyme

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Extraction of Enzymes Preparation of Crude Enzymes Purification of Enzymes Chromatography Adsorption Chromatography Ion Exchange Chromatography Gel Filtration Chromatography Affinity Chromatography Electrophoresis Immobilization of Enzymes or Whole Cells Techniques for Immobilization Adsorption Method Covalent Binding Method Cross Binding Method Some Reagents Used to Immobilize Enzymes by Cross Linking Entrapping Method Entrapping by Microencapsulation Requirements for Commercial use of Immobilized Enzyme Systems Enzyme Carrier Systems Some Applications of Immobilized Enzymes and Whole Cells Reactor Types Operating Strategy Stability of Immobilization and Protection of Immobilized EnzymesCells Some Examples of the Use of Immobilized Enzymes Cells Industrial Biocatalysis Biotransformation of Polyunsaturated Fatty Acids PUFAs Hydrolalse Mediated Biotransformations Oxidative Biotransformation Enzyme Engineering

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