Cross-linked enzyme aggregates (CLEA) are a type of immobilized enzyme obtained by precipitating enzymes from aqueous solutions and crosslinking the resulting aggregates using an appropriate crosslinking agent. When preparing CLEA particles, it is necessary to take into account many parameters, as the nature and amount of both precipitating and crosslinking agent depends on what the final product will be. It is also necessary to take into account the process parameters, which include temperature, pH and crosslinking time, as the enzymes differ from each other and it is necessary to determine the appropriate conditions for the preparation of CLEA particles. One of the key properties of CLEA particles is their size, on which transport phenomena depend and thus the availability of catalytic sites. With this kind of immobilization of enzymes, we can improve the activity, selectivity and stability of the enzyme and thus the success of the biocatalytic process. CLEA particles can also be obtained using magnetic nanoparticles to form magnetic CLEA particles. Different enzymes can be combined into combined cross-linked enzyme aggregates, which offer many advantages, especially in terms of production costs at the industrial level. By using CLEA particles in microflow reactors, we achieve the intensification of biocatalytic processes, which is the result of better heat and mass transfer and more controlled conditions in miniaturized devices.
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