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Enzymes may be immobilized by a variety of methods,
which may be broadly classified as physical, where weak interactions
between support and enzyme exist, and chemical,
where covalent bonds are formed with the enzyme.
To the physical methods belong: (i) containment of an enzyme
within a membrane reactor, (ii) adsorption (physical,
ionic) on a water-insoluble matrix, (iii) inclusion (or
gel entrapment), (iv) microencapsulation with a solid membrane,
(v) microencapsulation with a liquid membrane, and
(vi) formation of enzymatic Langmuir-Blodgett films. The
chemical immobilization methods include: (i) covalent attachment
to a water-insoluble matrix, (ii) crosslinking with
use of a multifunctional, low molecular weight reagent, and
(iii) co-crosslinking with other neutral substances, e.g. proteins.
Numerous other methods which are combinations of
the ones listed or original and specific of a given support
or enzyme have been devised. However, no single method
and support is best for all enzymes and their applications.
This is because of the widely different chemical characteristics
and composition of enzymes, the different properties of
substrates and products, and the different uses to which the
product can be applied. Besides, all of the methods present
advantages and drawbacks. Adsorption is simple, cheap and
effective but frequently reversible, covalent attachment and
crosslinking are effective and durable, but expensive and
easily worsening the enzyme performance, and in membrane
reactor-confinment, entrapment and microencapsulations
diffusional problems are inherent. Consequently, as a
rule the optimal immobilization conditions for a chosen enzyme
and its application are found empirically by a process
of trial and error in a way to ensure the highest possible
retention of activity of the enzyme, its operational stability
and durability.
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