This document discusses enzyme immobilization. It notes that immobilizing enzymes simplifies their use, enhances stability, allows for easy separation from products, reduces costs, and improves stability against various factors. Ideal support materials are described as being low cost, inert, stable, regenerable, and providing enhanced specificity and resistance to conditions. Various inorganic and organic solid supports are outlined, including their properties and example enzyme immobilizations. Methods of covalent bonding and applications of immobilized enzymes in areas like biosensing, medicine, food/dairy, and other uses are also summarized.

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

5. 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

7. 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.

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

9. 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

10. 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

11. 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

18. Methods of Covalent Bonding
1. Diazotization
2. Peptide Bond
3. Group activation
4. Poly-functional reagents

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

31. 2. Immobilized enzymes in medicine and antibiotic production

32. 3. Immobilized enzymes in Food and Dairy Industry

33. 4. Other applications of Immobilized enzymes