There is a wide variety of protocols for enzyme immobilization, allowing for the reuse of the enzyme, integration in flow bioreactors, and easy separation from the final product. However, none of them have reached a generalized implementation and new immobilization technologies are continuously being developed to improve the properties of the immobilized biocatalysts. In this chapter, we describe three advanced strategies looking at the key points of enzyme immobilization: the sustainability of the support, the recovered activity of the immobilized enzyme, and the reuse of the cofactors. Lignin is presented as a suitable and versatile support for enzyme immobilization, offering a more cost-effective and biodegradable strategy. A cationic polymer is used during the enzyme immobilization procedure to prevent the subunit dissociation of multimeric enzymes as well as to avoid excessive rigidification of the covalently immobilized enzyme. Finally, the reversible co-immobilization of cofactors has been improved by increasing the reactive groups of the support.
Keywords: Cofactor co-immobilization; Enzyme immobilization; Enzyme stability; Lignin; Polyethyleneimine.
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