Extraction of Enzymes, Tissue Homogenizer, Ultrasonic Vibrations, Ethylene Diamine Tetra Acetic Acid, Triton, Homogeneous

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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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Extraction of Enzymes
Fresh tissue is crushed into a paste with an extraction medium (often a buffer) in a mortar and pestle, or in a tissue homogenizer, or in a blender or by ultrasonic vibrations (sonication). The molarity and pH of the buffer is suitably adjusted (which may vary for different enzymes) to achieve maximum solubility and activity of the enzyme. EDT A (ethylene diamine tetra acetic acid) is often included in the extraction medium to remove heavy metals (which otherwise inhibit enzyme activity), and for disrupting the membranes of cells and cell organelles.

Detergents such as Triton-X are also used sometimes to solubilise the membranes. Many enzyme proteins contain disulfide (S-S) bonds due to the presence of cysteine residues, which are easily broken during enzyme extraction leading -to loss of enzyme activity. To overcome this problem are added, thiols such as-mercaptoethanol whose sulfhydryl (-SH) group is able to maintain the S-S linkage in enzymes.

If the extract is not homogeneous, the homogenate (extract) is filtered to remove cell debris, fibres, etc.; otherwise filtration may be avoided. All operations of extraction and purification are generally carried out in cold (0-4°C), since most of the enzymes get inactivated at higher temperatures.

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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