METHOD OF ENZYME IMMOBILIZATION & ITS APPLICATION

SUBJECT-BIOTECHNOLOGY

Presented by-

Puspendu sen
B. Pharm 3rd year 6th semester
Roll no-36501920004
Reg. No.-203650201910060
Session-2020-2024

ANAND COLLEGE OF EDUCATION

(DEPT. OF PHARMACY)
 CONTENT
 Introduction
 An ideal carrier materials for enzyme immobilization
 CLASSIFICATIONS OF CARRIERS
 Enzyme immobilization methods
 Physical method for immobilization
 Chemical method for immobilization
 Application of enzyme immobilization
 CONCLUSION
 Reference
 INTRODUCTION

 Enzyme Immobilization-
 Immobilized enzymes are enzymes which are attached to an inert, material, that will
prevent loss of enzyme activity by not changing chemical nature of the reactive groups in
the binding site of the enzyme.

Need For Immobilization-

 Re-use of enzymes for many reaction cycles

 Ability to stop the reaction rapidly by removing the enzyme from the reaction solution.
 Enhanced stability.
 Product is not contaminated with the enzyme.
 An Ideal Carrier Materials For Enzyme
Immobilization

 Inert.

 Physically strong and stable.

 Cost effective.

 Regenerable.

 Reduction in product inhibition.

 Enzyme Immobilization Methods
  Physical Method For
Immobilization

 Adsorption:
Involves the physical binding of the enzyme on
the surface of carrier matrix.
 Carrier may be organic or inorganic.
 The process of adsorption involves the weak
interactions like Vander Waal or hydrogen  Adsorption
bonds.
Carriers: silica, bentonite, cellulose, etc.
e.g. catalase & invertase
 CLASSIFICATIONS OF CARRIERS
 Inorganic Carriers •
High pressure stability.• May undergo abrasion.
.
Examples:1.Commercialy SiO 2.Porous glass.3. Silica.
 Organic Natural Carriers•
Favourable compatibility with proteins.

Examples:1.cellulose derivatives- i. DEAE-cellulose

2. Dextran.3.PolysacharidesAgarose, Starch.
 Organic Synthetic Carriers –
High chemical and mechanical stability.

Examples: 1.Polystyrene2.Polyvinylacetate3. Acrylic polymers.

  ENCAPSULATION
 In this method an enzyme is encapsulated within a capsule made up of semi permeable membranes like nitro
celluloses, nylon, hemi cellulosic structures etc.

 Advantages-
i. cheap and simple method.
ii. Immobilized large quantity of enzyme.

 Disadvantages-
i. pore size limitation.
ii. Only small substrate molecule is able to cross the
membrane.
 ENTROPMENT-
 In entrapment, the enzymes or cells are not directly attached to the support surface, but simply trapped
inside the polymer matrix.
 Inclusion in gels: Poly acrylamide gel
 Inclusion in fibres: Cellulose and Poly-acrylamide gels.
 Inclusion in micro capsules: Polyamine, Polybasic -acid chloride
monomers.
  Chemical Method For Immobilization
 Cross linking
 Cross linking involves intermolecular cross linking of enzyme molecules in the presence/absence of
solid support.
 The method produces a 3 dimensional cross linked enzyme aggregate
(insoluble in water) by means of a multifunctional reagent that links
covalently to the enzyme molecules.

 Advantages-
1.Very little desorption(enzyme strongly bound).
2. Higher stability (i.e. ph, ionic & substrate concentration).

 Disadvantages-
1. Cross linking may cause significant changes in the active site.
2. Not cost effective.
Cross linking
  Covalent Bonding
Ø Based on the binding of enzymes and water-insoluble
carriers by covalent bonds.
Ø The functional groups that may take part in this binding
are Amino group, Carboxyl group, Sulfhydryl group,
Hydroxyl group, Imidazole group, Phenolic group,
Thiol group, etc.

 Advantages-
 t provides an enzyme and a carrier with a strong binding
force.
 There is no leaking of enzymes.
 It’s a straightforward and popular method.

 Disadvantages-
 Different functional group assistance options are offered.
 Wide applicability.
.
  Application Of Enzyme Immobilization

 Industrial production:
Industrial production of antibiotics, beverages, amino acids. etc.

 Biomedical applications:
Immobilized enzymes are widely used in the diagnosis and treatment of many diseases.
Immobilization techniques are effectively used in drug delivery systems especially to oncogenic sites.

 Food industry: Enzymes like pectinases and cellulases immobilized on suitable carriers are
successfully used in the production of jams, jellies and syrups from fruits and vegetables.

 Research: A Research activity extensively uses many enzymes. The use of immobilized enzyme
allow researcher to increase the efficiency of different enzymes such as Horse Radish Peroxidase
(HRP) in blotting experiments and different Proteases for cell or organelle lysis.
  Application Of Enzyme Immobilization

(5). Production of bio-diesel from vegetable oils.

(6). Waste water management: -

treatment of sewage and industrial effluents.

(7). Textile industry: -

scouring, bio-polishing and desizing of fabrics.

8). Detergent industry:-

immobilization of lipase enzyme for effective dirt removal from cloths.
 CONCLUSION

Enzyme immobilization is one of the most promising

approaches for exploiting enzyme based processes in
Biotransformation, diagnostics, pharmaceutical and food
industries.

Several hundreds of enzymes have been immobilized in a

variety of forms including penicillin G Acylase, lipases,
proteases, invertase, etc.
 REFERENCE
 Pharmaceutical Biotechnology
By Dr. S.P. Vyas & Dr. V.K. Dixit
 Enzymes & its Immobilization Presentation
By Dr. S. Khanam
 Biotechnology text book
By U. Satyanarayana
Thank you