Enzyme Immobilization , Advantages And Disadvantages , Method Of Immobilization

1. Enzyme
Immobilization
Prepared By :-
Narayan Soni
B.Pharma 3rd Year
Mahakal Institute Of Pharmaceutical Studies , Ujjain

2. What Is Enzyme Immobilization
• Enzyme immobilization is a technique
specifically designed to restrict the freedom
of movement of an enzyme.

3. What Is An Immobilized Enzyme
• An Immobilized Enzyme Is One Whoes
Movement In Space Has Been Restricted
Either Completely Or To A Small Limited
Region

4. What Are The Advantages Of
Immobilized Enzyme
• Multiple Or Repetitive Use Of A Single Batch Of Enzymes.
• Continuous Production Systems Can Be Used , Which Is Not Possible
With Free Enzyme.
• Enzymes Can Be Used At Much Higher Concentrations Than Free
Enzyme.
• Immobilized Enzymes Are Usually More Stable.
• Ability To Stop The Reaction Rapidly By Removing The Enzyme From
The Reaction Solution.
• Product Is Not Contaminated With The Enzyme.
• Easy Separation Of The Enzyme From The Product.

5. What Are The Disadvantages Of Enzyme
Immobilization
• It Gives Rise To An Additional Bearing On Cost.
• It Invariably Affects The Stability And Activity Of Enzymes.
• This Approach Can Not be Used When One Of The Substrates Is
Insoluble.
• Immobiliztaion Requires Additional Time, Equipment.
• Any Contamination Is Costly To Deal With Because The Whole System
Would Need To Be Stopped.

6. Need For Immobilization
• Protection From Degradation And Deactivation.
• Recycling , Repitative Use.
• Cost Efficiency.
• Enhanced Stability.
• Use As Controlled Release Agents.
• The Ability To Stop The Reaction Rapidly By Removing The Enzyme From The
Reaction Solution.

7. Method Of Immobilization
• Based On Support Or Matrix And The Type Of Bonds Involved, There Are Five
Diffrent Method For Immobilization Of Enzyme Or Whole Cell.
1. Adsorption
2. Covalent Bonding
3. Entrapment
4. Cross Linking
5. Encapsulation

8. • Adsorption
In This Method Enzyme Is Adsorbed To External Surface Of The Support. The Support Or
Carrier Used May Be Of Different Types Such As :
1. Mineral Support ( Eg. Aluminium Oxide , Clay )
2. Organic Support ( Eg. Starch )
The Process Of Adsorption Involves The Weak Interaction Like Wander Waal Or
Hydrogen Bond

9. Advantage
• Simple And Economical
• Limited Loss Of Activity
• Can Be Recycled ,
Regenerated & Reuse
Disadvantage
• Relatively Low Surface Area
For Binding
• Exposure Of Enzyme To
Microbial Attack
• Efficiency Is Less

10. • Covalent Bonding
This Method Involves The Formation Of Covalent Bonds Between The Chemical Groups
In Enzyme And To The Chemical Groups On The Support Or Carrier.
• The Functional Groups That May Take Part In This Binding Are
Amino Group , Carboxyl Group , Hydroxyl Group , Imidazole Group , Phenolic Group ,
Thiol Group ETC.

11. • Disadvantages
Covalent Binding May Alter The Conformational Structure And Active Center Of The
Enzyme , Resulting In Major Loss Of Activity And / Or Changes Of The Substrate
• Advantages
The Binding Force Between Enzyme And Carrier Is So Strong That No Leakage Of The
Enzymes Occurs, Even In The Presence Of Substrate Or Solution Of High Ionic Strength

12. • Entrapment
• In Entrapment , The Enzymes Or Cell Are Not Directly Attached To The Support
Surface, But Simply Trapped Inside The Polymer Matrix
• Enzymes Are Held Or Entrapped Within The Suitable Gels or Fibers
Inclusion In Gels :- Poly Vinyl Alchol Gels
Inclusion In Fibers :- Cellulose
Inclusion In Microcapsule :- Polyamine

13. Advantage
• Fast Method Of
Immobilization
• Cheap ( Low Cost Matrixes
Available )
• Easy To Practice At Small
Scale
Disadvantage
• Leakage Of Enzyme
• Pore Diffusion Limitation
• Chance Of Microbial
Contamination

14. • Cross Linking
• In This Method Of Immobilization Enzymes Are Directly Linked By Covalent Bonds
Between Various Groups Of Enzymes Via Polyfunctional Reagents.
• Unlike Other Methods , There is No Matrix Or Support Involved In This Method.
• Commonly Use Polyfunctional Reagents Are
• Diazonium Salts
• Glutaraldehyde

15. • Disadvantages
1. Cross Linking May Cause Significant Changes In The Active Site
2. Not Cost Effective
• Advantages
1. Very Little Desorption ( Enzyme Strongly Bound )
2. Higher Stability

16. • Encapsulation
• This Type Of immobilization Is Done By Enclosing The Enzymes In A Membrane
Capsule.
• The Capsule Will Be Made Up Of Semi Permeable Membrane Like Nitrocellulose

17. Advantage
• Cheap And Simple Method
• Large Quantity Of Enzymes
Can Be immobilized by
Encapsulation.
Disadvantage
• Pore Size Limitation
• Only Small Substrate
Molecule Is Able to Cross
The Membrane.

18. Thank You
Prepared By :-
Narayan Soni
Reference :- 1. https://www.easybiologyclass.com/enzyme-cell-immobilization-techniques/
2. http://www.slideshare.net/ashwinimushunuri96/enzyme-immobilization-
53123282?from_m_app=android
3. http://www.slideshare.net/jahir143/enzyme-immobilization-
20940809?from_m_app=android