02753naa a2200385 a 450000100080000000500110000800800410001902400480006010000180010824501890012626000090031552015530032465000120187765000120188965000110190165000120191265300330192465300220195765300220197965300300200165300110203165300180204265300170206065300280207765300240210570000200212970000240214970000300217370000240220370000240222770000230225170000180227470000290229277300460232121764272025-06-04       2025    bl uuuu     u00u1 u    #d7 ahttps://doi.org/10.3390/ catal150200992DOI1 aCAMPOS, A. S.  aA New Multi-Active Heterogeneous Biocatalyst Prepared Through a Layer-by-Layer Co-Immobilization Strategy of Lipase and Laccase on Nanocellulose-Based Materials.h[electronic resource]  c2025  aLipase from Pseudomonas fl uorescens (PFL) and laccase from Trametes versicolor were co-immobilized onto nanocellulose (NC), using a layer-by-layer approach. Initially, PFL was adsorbed onto NC through ionic and hydrophobic interactions. To achieve higher PFL immobilization yield and activity, NC was functionalized with aldehyde groups through periodate oxidation (NCox) or glutaraldehyde activation (NC-GA). FTIR analysis confi rmed these chemical modifi cations. Among the functionalized NCs, NCox showed the best capacity to retain higher amounts of PFL (maximum load: 20 mg/g), and this support was selected to proceed with the co-immobilization experiments. In this process, NCox-250-PFL (NCox activated with 250 µmol/g of aldehyde groups) was covered with polyethyleneimine (PEI), laccase was co-immobilized, and a crosslinking step using glutaraldehyde was used to covalently attach the enzymes to the support, producing the biocatalyst NCox-250-PFL-PEI-Lac-GA. Co-immobilized enzymes presented higher thermal stability (50 ◦ C) than soluble enzymes; co-immobilized laccase retained 61.1% of its activity after 24 h, and PFL retained about 90% after 48 h of deactivation at 50 ◦ C. In operational stability assays, the heterogeneous biocatalysts maintained more than 45% of their activity after fi ve cycles of pNPB hydrolysis and ABTS oxidation. This co-immobilized biocatalyst, with its high stability and activity retention, is a promising multi-active heterogeneous biocatalyst for use in cascade reactions of industrial interest.  aEnzymes  aLaccase  aEnzima  aLípase  aAtivação de glutaraldeído  aCo-immobilization  aCo-imobilização  aGlutaraldehyde activation  aLacase  aNanocellulose  aNanocelulose  aOxidação de periodato  aPeriodate oxidation1 aJESUS, E. da C.1 aMENDES FILHO, P. F.1 aARAÚJO PEREIRA, A. P. DE1 aCASTRO, A. C. R. de1 aCOELHO, A. C. M. A.1 aBORDALLO, P. do N.1 aBORGES, W. L.1 aCARVALHO, A. C. P. P. de  tApplied Soil Ecology, 211, 106102 , 2025.