General Method of Enzyme

Immobilization and Its Carrier

Characteristics

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Introduction

Enzyme is an important biocatalyst with strong specificity, high catalytic efficiency, no

pollution, mild reaction conditions, etc. It has been widely used in pharmaceutical,
food, environmental protection, brewing, energy and other fields. However, in practical
applications, enzymes also have many shortcomings. For example, most enzymes are
easily deactivated and inactivated under the influence of external factors such as high
temperature, strong acid, strong alkali and heavy metal ions; they are mixed with
substrates and products. After the reaction, even if the enzyme still has high vitality, it is
difficult to recycle.

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Introduction

This single-use enzyme method not only increases the production cost, but also is
difficult to continuously produce; and the separation and purification are difficult,
which also leads to increase of production cost. The term immobilized enzyme was
recommended at the 1971 Enzyme Engineering Conference. With the development of
immobilization technology, immobilized cells appear. In 1973, for the first time in Japan,
the aspartase in immobilized E. coli cells was industrially applied, and L-aspartate was
continuously produced from fumaric acid. Immobilized enzyme technology provides an
effective means for solving these problems, and thus becomes one of the most active
research directions in the field of enzyme engineering.

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1. Traditional methods of
enzyme immobilization

The key is to select the appropriate immobilization method

and the necessary carrier.

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1.1 Adsorption method

The adsorption method is a method of immobilizing an enzyme on a carrier such as

cellulose, agarose or a carrier such as porous glass or ion exchange resin by a physical
adsorption method. The remarkable features are: simple process and mild conditions,
including inorganic and organic polymer materials, the adsorption process can
simultaneously achieve purification and immobilization; the enzyme can be reactivated
after inactivation, and the carrier can also be regenerated. However, the carrier is
required to have a large specific surface area and a lively surface.

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1.2 Embedding method

The embedding immobilization method is to immobilize the enzyme in a

lattice structure such as a lattice structure or a microcapsule structure of a
polymer material, and the substrate can still penetrate into the lattice or the
microcapsule to contact the enzyme. This method is relatively simple, for the
enzyme molecule is only embedded, and the degree of destruction of
biological activity is low, but this method is not applicable to
macromolecular substrates.

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1.3 Combination method

A method of immobilizing an enzyme by ion bonding on the surface of an

enzyme protein molecule and an insoluble solid phase support is called an
ionic bond method. The immobilization method in which chemical covalent
bond formation is formed is called covalent bond bonding. The covalent
bond method has strong binding force, and the enzyme is not easily
detached during use, and the stability is good. The disadvantage of this
method is that the activation or immobilization of the carrier is complicated
and the reaction conditions are relatively strong, so it is often necessary to
strictly control the conditions to obtain the immobilized enzyme with higher
vigor.

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1.4 Cross-linking method

The cross-linking method uses a multifunctional reagent to crosslink

between enzyme proteins, so that a covalent bond is formed between the
enzyme molecule and the multifunctional reagent, and a three-way cross-
linking structure is obtained, except that cross-linking occurs between the
enzyme molecules. There is also some intramolecular crosslinking.

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