Immobilization of enzyme

Represented to:
Dr. Syed Aun Muhammad

Represented By:
Kiran jameel
Zainab Akhtar
Huma Mujahid
Iqra Aziz
Kauser Parveen

22 (M)
20 (E)
11(M)
05(E)
22(E)

What is immobilization?

The act of limiting movement or making

incapable of movement.

Introduction
Immobilization

of enzymes can be defined as

the Confinement of enzymes (bio-catalyst) in a
distinct phase, separated from the bulk phase
but allowing it to exchange with the latter.
Bulk phase consists of a substrate, an effectors
or inhibitor.
Immobilized enzymes is either Physically
entrapped or covalently bonded by Chemical
means to an inert insoluble matrix or carrier in
other words, it involves the restrictive
localization of enzymes.
Matrix is generally High molecular weight
polymer

Need for immobilization

Accelerates
Specificity
Cost
Not

the chemical reactions

and un-modified enzymes

effective

difficult to separate

Attachment

to polymers/matrix , causes re-use.

Carrier for immobilized

enzymes

Ideal characteristics of the carrier

Low cost and of optimum quality.

Inertness
Physical

strength

Stability
Regenerability
Enhancement

of enzyme specificity
Reduction of product inhibition

Types of carrier

Naturally

occurring
Structural proteins(Ex, ceratin

,collagen)
Globular proteins (Ex, albumin)
Carbohydrates (Ex, dextran)

Synthetic organic
Ex Polyvinyl, Epoxide etc

Inorganic
Ex glass, silicate, bentonite,titania

etc.

Methods of Immobilization

Physical Methods

Adsorption
Entrapping
Membrane
confinement

Chemical Methods

Covalent bonding
Cross linking
Complexation and
chelation

Parameters for method

selection
Overall

catalytic activity
Effectiveness of the catalytic utilization
Deactivation and regeneration characteristics
Cost effective
Intended application of immobilized enzymes
Toxicity of immobilized enzyme
Waste disposal

Physical Methods
Adsorption
Non-specific bonding like electrostatic or hydrophobic affinity
binding to special ligand
Mostly explained in following term:
Static pores
Dynamic pores
Reactor loading
Electro depression

Advantages
Simple & economical
Limited loss of activity
Can be recycled and regenerated & reused

Disadvantages
Relatively low surface area for bonding
Exposure of enzyme to microbial attack
Smaller particles cause high pressure drop in
continuous
Packed bed reactor
Yield are often low due to inactivation &
desorption

Entrapping
Enzymes are held or entrapped within the suitable gels or fibre
In a gel it may causes:
Matrix polymerization
Precipitation
Coagulation
Entrapment in calcium alginate is the most widely used for
entrapment for:
Microbial
Animal
Plant enzyme/or cell
Ex: Glucose oxidase + polyacrlamide