Via Immobilization Techniques

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Stabilization by random immobilization

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Rigidification of 3D structure

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Stabilization of multimeric enzymes

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Chemical modification of immobilized enzymes

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Generation of hyper-hydrophilic microenvironments

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Hyper-activation of lipases

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Modulation of enantio-selectivity of lipases

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Reduction of inhibitions

Fig. 3. Possible use of immobilization techniques to improve enzyme properties.

Fig. 3. Possible use of immobilization techniques to improve enzyme properties.

immobilized enzymes with regard to soluble enzymes, as soluble enzymes are able to undergo aggregations and interactions with hydrophobic interfaces.

4.2. Stabilization of Enzymes by Multipoint Covalent Immobilization

Multipoint covalent attachment of enzymes on highly activated pre-existing supports (see Fig. 4), via very short spacer arms and through a number of residues on the enzyme surface, can promote dramatic stabilization of the three-dimensional (3D) structure of the immobilized enzyme. In this case, the relative distances among all residues involved in multipoint immobilization have to be maintained unaltered during any conformational change induced by a distorting agent (e.g., heat, organic cosolvents, extremes of pH values). In this way, the intensity of conformational changes involved in enzyme inactivation may be strongly reduced and the immobilized enzyme may become strongly stabilized. In fact, a number of enzymes have been dramatically stabilized by multipoint cova-lent immobilization as compared with one-point immobilized counterparts (17-20).

4.3. Stabilization of Multimeric Enzymes by Multisubunit Immobilization

Some multimeric enzymes, under certain experimental conditions, may be inactivated via dissociation of the subunits of their quaternary structure. Some protocols for enzyme immobilization may prevent dissociation of subunits and may therefore result in dramatic stabilizing effects (21).

4.3.1. MultiSubunit Covalent Attachment on Pre-Existing Supports

Both subunits of dimeric enzymes may be easily attached to the support when very highly activated supports are used, resulting in very important stabilizing effects (22). Of greater difficulty is the stabilization of more complex multimeric enzymes. In this case, several subunits (two or three) can be covalently attached to

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