ENZYME BIOTECHNOLOGY.pptx

Enzyme immobilization

1. ENZYME IMMOBILIZATION BY- Ms. Aishwarya Gangwar Assistant Professor MMCP, Mullana , Ambala

2. Immobilized enzymes simplify the manipulation with the biocatalyst and the control of the reaction process. Enhance the stability of the enzymes under both storage and operational conditions. Immobilization provides an effortless separation of the enzymes from the product and hence protein contamination of the product is minimized or avoided altogether. Enzyme immobilization remarkably reduces the cost of enzyme and the enzymatic products. Immobilization improves the stability of enzymes against temperature, solvents, pH, contaminants and impurities. Immobilization helps for efficient recovery and reuse of expensive enzymes. Advantages of Immobilized Enzymes over Conventional Enzymes

3. Ideal Properties of Support materials used in Enzyme Immobilization It should be low cost and eco-friendly, reducing the economic impact of the process. It should be totally inert after immobilization and not blocking the desired reaction. It should be highly stable. It should have high regenerability after the useful lifetime of the immobilized enzyme. It should enhance the enzyme specificity. It should have thermal and mechanical resistance, allowing the immobilized enzyme to be used under various operational conditions.

4. SOLID SUPPORT Inorganic Glass, Silica gel, Alumina, Metal oxides, Ceramica Organic SYNTHETIC Polystyrene, Silk, Polyvinyl chloride NATURAL Alginate, Chitosan, Collagen, Gelatin, Starch

5. Inorganic Support material Inorganic Support materials are widely used for immobilization as they have thermal and mechanical resistance. They also offer microbial resistance as inorganic supports does not serve as substrate for bacteria/fungal growth. The characteristic feature of inorganic support is they provide rigidity and porosity. Furthermore, they ensure invariance of pore diameter/pore volume which ensures fixed volume and shape to the support . Inorganic Material Properties Enzyme Immobilized Zeolite Large specific surface area of the zeolite substrate resulted in high enzyme loading Glucose oxidase, lysozyme Glass Highly viscous liquid α - Amylase, nitrite reductase Activated carbon Large contact sites, high surface area Acid protease, acidic lipase Charcoal Excellent adsorbent Papain, amyloglucosidase

6. Organic Support material 1. Natural supports: A wide variety of natural polymers, mainly water-insoluble polysaccharides have been used as support matrix for enzyme immobilization. The characteristic features of these polymers that make them good support system include, their ability to form inert gels, due to their chemical structure which can be achieved easily, they can bind to proteins and enzymes in a reversible and irreversible way, they are available in large quantities, inexpensive and they show high thermal and mechanical resistance by cross-linking with bifunctional reagents. Natural Material Properties Enzyme Immobilized Alginate Used as Xanthan-alginate beads, alginate- polyacrylamide gels, improve the stability of enzymes Protease, Pectinase Collagen Retain significant activity after many cycles of reuse Tannase, catalase Starch stable Bitter gourd peroxidase Chitosan and chitin More reliable for enzyme trapping D- Hydantoinase

7. 2. Synthetic supports: Synthetic polymers are ion-exchange resins which have porous surface and are insoluble in nature. These polymers, because of their porous structure, can immobilize the enzyme very strongly. They are inert to microbial attack. Synthetic Material Properties Enzyme Immobilized Polyvinyl chloride Prevent thermal inactivation of the enzyme Cyclodextrin glucosyltransferase Polyurethane microparticles Improve enzyme loading and efficiency Polyethylene glycol Forms a protective layer around the active center of enzyme to prevent the effect of oxidative stress White radish peroxidase UV- activated polyethylene glycol High porosity used in waste water treatment Glutaraldehyde-activated nylon lipase

8. Methods of Covalent Bonding Diazotization Peptide Bond Group activation Poly-functional reagents

9. Applications of Immobilized enzymes 1. Immobilized enzymes as biosensors

10. 2. Immobilized enzymes in medicine and antibiotic production

11. 3. Immobilized enzymes in Food and Dairy Industry

12. 4. Other applications of Immobilized enzymes