Presentation on immobilization of enzymes in a very simplified manner and easy language, can help to clearing of concepts and may also be used as reference for any presentation,seminar or assignment
Presentation on immobilization of enzymes in a very simplified manner and easy language, can help to clearing of concepts and may also be used as reference for any presentation,seminar or assignment
Presentation on immobilization of enzymes in a very simplified manner and easy language, can help to clearing of concepts and may also be used as reference for any presentation,seminar or assignment
Presentation on immobilization of enzymes in a very simplified manner and easy language, can help to clearing of concepts and may also be used as reference for any presentation,seminar or assignment
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PRESENTED BY
SHIRJEEL AHMAD SIDDIQUI
B.Sc. Biotechnology Semester VI Enzymes are biocatalyst synthesized by living cells. They are complex protein molecules that bring about chemical reactions concerned with life. They can be produced in large amounts by microorganisms for industrial applications. Enzymes in free state react with substrates to result in products. Such use of enzymes is wasteful particularly for industrial purposes ,since they are not stable and can’t be recovered or reused. Immobilization of enzymes refers to the technique of confining/anchoring the enzymes in or on an inert support for their stability and functional reuse. By employing this technique, enzymes are made more efficient and cost effective for their industrial use. Some workers regard immobilization as “a goose with a golden egg” in enzyme technology It involves the physical binding of enzymes on the surface of an inert support. The support materials may be inorganic (alumina, silica gel, calcium phosphate gel) or organic (starch, carboxymethyl cellulose,DEAE) Adsorption of enzymes involves weak forces like van der Waals forces and hydrogen bonds. Therefore, adsorbed enzymes can be easily removed by minor changes in pH or temperature. Sometimes this feature also serve as disadvantage for industrial use. Enzymes can be immobilized by physical entrapment inside a polymer or gel matrix. The size of the matrix pores is such that the enzyme is retained while substrate and product molecule is passed through. The matrices used for entrapping enzymes are made of polyacrylamide gel ,collagen, gelatin, starch, cellulose, silicone and rubber. The major limitation for entrapment of enzymes is their leakage from matrix. It is used in industrial production of amino acids. It is also considered as a type of entrapment. It refers to the process of spherical particle formation wherein a liquid or suspension is enclosed in semi- permeable membrane . The membrane may be polymeric, lipoidal, or non-ionic in nature. There are three distinct ways of encapsulation - a) Building of special membrane reactors. b) Formation of emulsions. c) Stabilization of emulsions to form microcapsule. It involves the formation of covalent bonds between the chemical group of enzymes and chemical group of support. It can be achieved by forming the bonds commonly specific as per the support and enzyme, it includes – a) Cyanogen bromide activation - glycol groups of support materials are activated by CNBr and binds to enzyme. b) Diazotation – amino derivatives support is treated with NaNO2 and HCl which in turn bind covalently to tyrosyl or histidyl group of enzymes. c) Peptide bond formation – The amino or carboxyl group of support binds with carboxyl or amino groups of enzyme respectively to form peptide bonds The enzyme molecules are immobilized by forming cross- linking, through the involvement of polyfunctional reagents. These reagents react with enzyme molecules and create bridges which form the backbone to hold enzymes molecules. Several reagents used in cross-linking like glutaraldehyde, diazobenzidine, hexamethylene disisocyanate. The selection of a particular method for immobilization of enzymes is based on trial and error approach to choose the ideal one. The major factors to be considered are as follows – Enzyme catalytic activity Stability Regenerability Cost factor The enzyme which have to be used in soluble form e.g. enzymes in food additives, liquid detergents and diagnostic reagents. They are stabilized using additives and chemical methods, they include – Solvent stabilization Substrate stabilization Stabilization by polymers Stabilization by salts Stabilization by chemical modifications In food and beverage industry In biochemical analysis In affinity chromatography and purification Improved drug delivery Immobilization of artificial cells/organs Stable and more efficient in function. Can be reused many times. Products are enzyme free. Control of enzyme function is easy Suitable for industrial and medical use Minimize effluent disposal problems Being so advantageous it has some disadvantages too – The possibility of loss of biological activity of an enzyme during its immobilization. Not suitable for multiple enzymatic reactions. Expensive affair often requires sophisticated equipment. Pages 286-290, Enzyme Technology, Biotechnology, U.Satyanaryana, Tenth edition,2016 Image courtesy via internet – https://www.google.co.in/search?q=immobilized+enzymes +and+their+products+table&source=lnms&tbm http://www.easybiologyclass.com/wp- content/uploads/2015/04/Enzyme-whole-cell- immobilization-methods-easybiologyclass.jpg https://www.google.co.in/search?q=images+of+adsorption +in+immobilisation+of+enzymes&source=lnms&tbm=isc h&sa=X&ved=