Multienzyme Coimmobilization on Triheterofunctional Supports

• Published in: Biomacromolecules Vol. 24; no. 2; pp. 929 - 942
• Main Authors: Santiago-Arcos, Javier, Velasco-Lozano, Susana, López-Gallego, Fernando
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 13.02.2023
• Subjects: Aldehydes; Biocatalysis; Catalysis; Enzymes, Immobilized - chemistry
• ISSN: 1525-7797; 1526-4602
• DOI: 10.1021/acs.biomac.2c01364

Immobilized multienzyme systems are gaining momentum in applied biocatalysis; however, the coimmobilization of several enzymes on one carrier is still challenging. In this work, we exploited a heterofunctional support activated with three different chemical functionalities to immobilize a wide variety of different enzymes. This support is based on agarose microbeads activated with aldehyde, amino, and cobalt chelate moieties that allow a fast and irreversible immobilization of enzymes, enhancing the thermostability of most of the heterogeneous biocatalysts (up to 21-fold higher than the soluble one). Furthermore, this trifunctional support serves to efficiently coimmobilize a multienzyme system composed of an alcohol dehydrogenase, a reduced nicotinamide adenine dinucleotide (NADH) oxidase, and a catalase. The confined multienzymatic system demonstrates higher performance than its free counterpart, achieving a total turnover number (TTN) of 1 × 105 during five batch consecutive cycles. We envision this solid material as a platform for coimmobilizing multienzyme systems with enhanced properties to catalyze stepwise biotransformations.