131818 20241125101202.0 doi 10.1021/acssuschemeng.3c02958 sideral 137222 ART-2023-137222 eng Diamanti, Eleftheria Self-Sufficient Heterogeneous Biocatalysis through Boronic Acid-Diol Complexation of Adenylated Cofactors 2023 Access copy available to the general public Unrestricted Self-sufficient heterogeneous biocatalysts (ssHB) are promising candidates for implementing cofactor-dependent enzymes in chemical biomanufacturing. Most strategies for coimmobilizing cofactors with dehydrogenases on porous agarose microbeads involve the use of cationic polymers (i.e., polyethylenimine, PEI) that interact electrostatically with the phosphate groups of their corresponding phosphorylated cofactors. Although the latter is a powerful and versatile approach, ionic bonds are disrupted in biotransformations operating at a high ionic strength, where screening of bonded ions takes place. Harnessing the ribose groups present in adenylated cofactors, we immobilize a selection of these (NAD(P)H, NAD(P)+, FAD, and ATP) on agarose microbeads functionalized with boronic acid to establish reversible covalent bonds between the cis-diol of the ribose in the cofactor backbone and the boronic acid. To do so, we functionalize cobalt-activated porous agarose beads with boronic acid (AG-B/Co2+) for the coimmobilization of adenylated cofactors with the corresponding cofactor dependent His-tagged dehydrogenases. First, we demonstrate that the adenylated cofactor-support interactions are reversible but show resistance against high salt concentrations, overcoming the main limitation of the current self-sufficient heterogeneous biocatalysts. Then, we coimmobilized several His-tagged enzymes with their corresponding adenylated cofactors and investigated the functionality and stability of these ssHBs in reductive aminations performed under high ionic strength in both batch and flow reactors. As a result, we manage to reuse the immobilized enzymes and the cofactors 3.5 × 105 and 167 times, respectively. This work expands the usefulness of ssBHs for hitherto bioprocess regardless of the ionic strength of the media. info:eu-repo/grantAgreement/EC/H2020/818089/EU/Artificial metabolic cells for biomanufacturing of bio-based chiral fine chemicals/METACELL This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 818089-METACELL info:eu-repo/grantAgreement/ES/MICINN/MDM-2017-0720 info:eu-repo/semantics/openAccess All rights reserved http://www.europeana.eu/rights/rr-f/ 7.1 2023 ENGINEERING, CHEMICAL 21 / 170 = 0.124 2023 Q1 T1 GREEN & SUSTAINABLE SCIENCE & TECHNOLOGY 21 / 91 = 0.231 2023 Q1 T1 CHEMISTRY, MULTIDISCIPLINARY 43 / 231 = 0.186 2023 Q1 T1 1.664 2023 Environmental Chemistry 2023 Q1 Renewable Energy, Sustainability and the Environment 2023 Q1 Chemistry (miscellaneous) 2023 Q1 Chemical Engineering (miscellaneous) 2023 Q1 13.8 2023 info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion Velasco-Lozano, Susana (orcid)0000-0001-9478-6750 Grajales-Hernández, Daniel Orrego, Alejandro H. Di Silvio, Desiré Fraile, José María (orcid)0000-0002-0136-5138 López-Gallego, Fernando 11, 39 (2023), 14409-14421 ACS sustain. chem. & eng. ACS Sustainable Chemistry and Engineering 2168-0485 858920 http://zaguan.unizar.es/record/131818/files/texto_completo.pdf Postprint info:eu-repo/date/embargoEnd/2024-09-21 2625931 http://zaguan.unizar.es/record/131818/files/texto_completo.jpg?subformat=icon icon Postprint info:eu-repo/date/embargoEnd/2024-09-21 oai:zaguan.unizar.es:131818 articulos driver 2024-11-22-12:12:27 ARTICLE