Scribd
Upload
60 views22 pages

Enzyme Immobilization Methods and Its Applications

Uploaded by

minhsmunadahmad
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
60 views22 pages

Enzyme Immobilization Methods and Its Applications

Uploaded by

minhsmunadahmad
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
You are on page 1/ 22

Enzyme

immobilization
methods and its
applications
What is enzyme immobilization?

• Immobilization is defined as the imprisonment of


cell or enzyme in a distinct support or matrix.

• Immobilized enzymes are enzymes which are


attached in or onto the surface of an insoluble
support

• The first immobilized enzyme was amino acylase


for the production of L­-amino acids in Japan.
Supports or Matrix used in
immobilization technology:

grouped into three major categories


(1). Natural polymers
e.g Alginate, collagen, Carrageenan, cellulose ,chitin,
starch
(2). Synthetic polymers
e.g Diethylaminoethyl cellulose (DEAE cellulose),
Polyvinyl chloride (PVC), UV activated Polyethylene
glycol (PEG)
(3). Inorganic materials
Advantages of immobilized enzymes:
(1). Increased functional efficiency of enzyme
(2). Enhanced reproducibility of the process
(3). Continuous use of enzyme
(4). Less labour input in the processes
(5). Saving in capital cost and investment of the
process
(6). Minimum reaction time
(7). Less chance of contamination in products
(8). More stability of products
(9). Improved process control
(10). High enzyme substrate ratio
Disadvantages of enzyme immobilization:
(1). High cost for the isolation, purification and
recovery of active enzyme
(2). Industrial applications are limited and only very
few industries are using immobilized enzymes
(3). Catalytic properties of some enzymes are reduced
or completely lost after immobilization on support or
carrier.
(4). Some enzymes become unstable after
immobilization.
Enzyme Immobilization Methods
(1). Adsorption
(2). Covalent bonding
(3). Entrapment
(4). Copolymerization
(5). Encapsulation
(1). Adsorption
• The oldest and simplest method of enzyme
immobilization.
• Nelson & Griffin used charcoal to adsorb invertase
for the first time in 1916.
• In this method enzyme is adsorbed to external
surface of the support.
• The support or carrier used for adsorption:
(1). Mineral support (Eg. aluminum oxide, clay)
(2). Organic support (Eg. starch)
(3). Modified sepharose and ion exchange resins
• There is no permanent bond formation between
carrier and the enzyme in adsorption method. Only
weak bonds stabilize the enzymes to the support
or carrier
(2). 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.

• It is one of the widely used methods of


enzyme immobilization.
Carriers or supports commonly used for
covalent bonding are:
(a). Carbohydrates: Eg. Cellulose, DEAE cellulose,
Agarose
(b). Synthetic agents: Eg. Polyacrylamide
(c). Protein carriers: Collagen, Gelatin
(d). Amino group bearing carriers: Eg. amino benzyl
cellulose
(e). Inorganic carriers: Porous glass, silica
(f). Cyanogen bromide (CNBr)­agarose and CNBr
Sepharose
(3). Entrapment:
• In this method enzymes are physically entrapped
inside a porous matrix.
• Bonds involved in stabilizing the enzyme to the
matrix may be covalent or non­covalent
• Examples of commonly used
matrixes for entrapment are:
(1). Polyacrylamide gels
(2). Cellulose triacetate
(3). Agar
(4). Gelatin
(5). Carrageenan
(6). Alginate
(4). Cross linking (copolymerization):
• This method is also called as copolymerization.
• Immobilization of 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 used polyfunctional reagents are
glutaraldehyde and diazonium salt.
(5). 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 nitro cellulose or nylon.
• In this method the effectiveness depends upon the
stability of enzymes inside the capsule.
Applications of enzyme immobilization:
(1). Industrial production:
• Production of antibiotics, beverages, amino acids
(2). Biomedical applications:
• In the diagnosis and treatment of many diseases.
• To overcome inborn metabolic disorders by the supply
of immobilized enzymes.
• effectively used in drug delivery systems especially to
oncogenic sites.
(3). Food industry:
Enzymes like pectinases and cellulases immobilized
on suitable carriers used in the production of jams,
jellies and
(4). Research:
used to increase the efficiency of different enzymes
such as Horse Radish Peroxidase (HRP) in blotting
experiments and different Proteases for cell or
organelle lysis.
(5). Waste water management:
treatment of sewage and industrial effluents.
(6). Textile industry: scouring, bio­polishing
and desizing of fabrics.
(7). Detergent industry: immobilization of
lipase enzyme for effective dirt removal from
cloths.

You might also like