Immobilization of enzyme ppt

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Basics and various methods of Enzyme Immobilization

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By: drbyog (104 month(s) ago)

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By: drbyog (104 month(s) ago)

nice ppt for student point of view

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IMMOBILIZATION OF ENZYME BY » N.V. Chandak

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Enzymes are protein molecules which serve to accelerate the chemical reactions of living cells. The term “ immobilized” means unable to move or stationary .  And that is exactly what an immobilized enzyme is :  an enzyme that is physically attached to a solid support over which a substrate is passed and converted to product.

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Carrier-Binding

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Carrier-Binding The selection of the carrier depends on the nature of the enzyme itself Surface area Particle size Molar ratio of hydrophilic to hydrophobic groups Chemical composition Physical Adsorption Ionic Binding Covalent Binding TYPES α -Amylase adsorbed on Calcium phosphate EX EX Polysaccharides and synthetic polymers EX Cellulase to Polyurethane by Isocynate Glucose oxidase to Porous glass by Isothiocyanate

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Cross-Linking Immobilization of enzymes has been achieved by intermolecular cross-linking of the protein, either to other protein molecules or to functional groups on an insoluble support matrix . Generally, cross-linking is best used in conjunction with one of the other methods . For example » carbamy phosphokinase cross-linked to alkyl amine glass with glutaraldehyde .

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Entrapping Enzymes

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Lattice-Type It involves entrapping enzymes within the interstitial spaces of a cross-linked water-insoluble polymer. Some synthetic polymers such as polyarylamide, polyvinylalcohol, etc... and natural polymer (starch) have been used to immobilize enzymes using this technique. Microcapsule-Type Entrapping involves enclosing the enzymes within semi permeable polymer membranes. The preparation of enzyme micro capsules requires extremely well-controlled conditions.

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Procedures for micro capsulation Interfacial Polymerization : In this procedure, enzymes are enclosed in semi permeable membranes of polymers.

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Liquid Drying : In this process, a polymer is dissolved in a water-immiscible organic solvent which has a boiling point lower than that of water. An aqueous solution of enzyme is dispersed in the organic phase to form a first emulsion of water-in-oil type. The first emulsion containing aqueous micro droplets is then dispersed in an aqueous phase containing protective colloidal substances such as gelatin.

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Phase Separation : One purification method for polymers involves dissolving the polymer in an organic solvent and re-precipitating it.  This is accomplished by adding another organic solvent which is miscible with the first, but which does not dissolve the polymer.

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Applications of immobilized enzyme Pharmaceutical Isomerization of glucose to fructose (High fructose syrup) Lactose hydrolysis (dairy products converted to avoid lactose intolerance) Penicillin acylase

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Clinical BIOREMEDIATION DETECTION OF HEROIN AND ITS METABOLITES

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