Immobilized enzyme
An immobilized enzyme is an enzyme which is attached to an inert, insoluble material such as calcium alginate (produced by reacting a mixture of sodium alginate solution and enzyme solution with calcium chloride). This can provide increased resistance to changes in conditions such as pH or temperature. It also allows enzymes to be held in place throughout the reaction, following which they are easily separated from the products and may be used again – a far more efficient process and so is widely used in industry for enzyme catalysed reactions. An alternative to enzyme immobilization is whole cell immobilization.
In 1916, Nelson and Griffin discovered that invertase xhibited the same activity when absorbed on a solid (charcoal or aluminium hydroxide) at the bottom of the reaction vessel as when uniformly distributed throughout the solution. This discovery was the first of various enzyme immobilization techniques currently available.
Besides adsorption, different covalent methods of enzyme immobilization were developed in the 1950s and 1960s. Up to now, more than 5000 publications and patents have been published on enzyme immobilization techniques. Several hundred enzymes have been immobilized in different forms and approximately a dozen immobilized enzymes, for example penicillin G acylase, lipases, proteases, invertase, etc. have been used as catalysts in various large scale processes.
Commercial use
Immobilised enzymes are very important for commercial uses as they possess many benefits to the expenses and processes of the reaction of which include:
Convenience: Minuscule amounts of protein dissolve in the reaction, so workup can be much easier. Upon completion, reaction mixtures typically contain only solvent and reaction products.
Economical: The immobilized enzyme is easily removed from the reaction making it easy to recycle the biocatalyst.
Stability: Immobilized enzymes typically have greater thermal and operational stability than the soluble form of the enzyme.
this is also known as enzyme mobility
Immobilisation of an Enzyme
There are three different ways in which one can immobilise an enzyme which are the following, and are listed in order of effectiveness:
Adsorption on glass, alginate beads or matrix: Enzyme is attached to the outside of an inert material. Generally speaking, this method is the slowest among those listed here. As adsorption is not a chemical reaction, the active site of the immobilized enzyme may be blocked by the matrix or bead, greatly reducing the activity of the enzyme.
Entrapment: The enzyme is trapped in insoluble beads or microspheres, such as calcium alginate beads. However, this insoluble substances hinders the arrival of the substrate, and the exit of products.
Cross-linkage: The enzyme is covalently bonded to a matrix through a chemical reaction. This method is by far the most effective method among those listed here. As the chemical reaction ensures that the binding site does not cover the enzyme’s active site, the activity of the enzyme is only affected by immobility. However the inflexibility of the covalent bonds precludes the self-healing properties exhibited by chemoadsorbed self-assembled monolayers. Use of a spacer molecule like poly(ethylene glycol) helps reduce the steric hindrance by the substrate in this case.
External links
Article on enzyme immobilization.
v d e
Concepts in asymmetric synthesis
Chirality types
Chirality Stereocenter Planar chirality Chiral ligand Axial chirality Supramolecular chirality Inherent chirality
Chiral molecules
Stereoisomer Enantiomer Diastereomer Meso compound Enantiomeric excess Diastereomeric excess
Analysis
Optical rotation Chiral derivatizing agents NMR spectroscopy of stereoisomers Ultraviolet-visible spectroscopy of stereoisomers
Chiral resolution
Crystallization Kinetic resolution Chiral column chromatography Diastereomeric recrystallization
Reactions
Asymmetric induction Chiral pool synthesis Chiral auxiliaries Asymmetric catalysis Organocatalysis Biocatalysis
Categories: Enzymes | Organic chemistry | Stereochemistry
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