Lipase immobilization via cross-linked enzyme aggregates: Problems and prospects – A review

(Magnetic) Cross-Linked Enzyme Aggregates of Cellulase from T. reesei: A Stable and Efficient Biocatalyst

“…To achieve such reusable, thermostable biocatalysts with improved activity, cellulase CLEAs and mCLEAs were parallelly synthesized. Immobilization through CLEAs and mCLEAs synthesis involves the entire surface of the enzyme in the immobilization and presents some advantages in comparison with more traditional immobilization methods, such as easy and simple preparation [ 14 , 20 ]. It also exhibits high catalyst productivities, which were confirmed in our studies with improved expressed activities through careful optimization of parameters.…”

“…Extensive research was performed by Fernandez-Lafuente et al regarding stabilization of enzymes via immobilization [ 13 , 14 , 15 , 16 , 17 , 18 , 19 ]. Enzyme immobilization must be designed not only to solve the problem of enzyme recovery and reusability, but also to improve enzymes features, such as stability, activity, specificity, and selectivity.…”

“…Furthermore, a review by Fernandez-Lucas shows that CLEAs in anhydrous medium behave like hard particles, while when in aqueous medium, their consistency is very poor, similar to gelatin [ 36 ]. As suggested in a review by Sampaio et al [ 14 ], another possibility to increase CELAs stability is to use a feeder that is rich in amino groups, such as a protein or an aminated polymer [ 37 , 38 , 39 ]. Such a solution is efficient; however, it means the feeder increases the cost-related aspects and therefore decreases the advantages of CLEAs in comparison with other immobilization methods.…”

Engineering ketoreductases for the enantioselective synthesis of chiral alcohols

“…Cross-linked enzyme aggregates (CLEAs) are a wellknown example of the latter. 40,41 A variety of reactive functional groups inherent to enzymes, such as NH 2 , SH and OH, can be used to affect cross-coupling. Moreover, the introduction of noncanonical amino acids (ncAAs), bearing highly reactive groups such as N 3 , CRCH and NC, through genetic coding, significantly expands the number of functional groups that can be introduced.…”

Co-Immobilizing Two Glycosidases Based on Cross-Linked Enzyme Aggregates to Enhance Enzymatic Properties for Achieving High Titer Icaritin Biosynthesis

“…CLEAs have attracted much attention in recent years because of the high efficiency, great specificity, and easy operation. , The development of CLEAs mainly consists of two steps: first to precipitate the required enzyme by a precipitating agent (ammonium sulfate or any organic solvent) and then cross-link the aggregates via bifunctional reagents . Different from the carrier-based immobilization such as nanoparticle adsorption, metal-organic frameworks, biotin–avidin interactions, and so forth, the preparation of CLEAs does not require the intervention of carriers, thus avoiding the inactivation of enzymes due to the compatibility problem between carriers and target enzymes. − In addition, this technology could enable the enzyme to be immobilized directly from the crude enzyme solution without purification, which greatly reduces the cost of industrial application . Unfortunately, cross-linking is not easy to occur when the protein is poor on Lys.…”