3D‐Printed Platforms for Enzyme Immobilisation Screening and Direct Translation into Continuous‐Flow Biocatalysis

The development of immobilised biocatalysts for continuous-flow processes is still largely based on laborious trial-and-error screening, and conditions optimised on conventional carriers are often not directly transferable to structured reactors. Here, we introduce a 3D-printed methodology that integrates high-throughput screening of enzyme immobilisation with implementation in a continuous-flow reactor using the same photopolymeric formulation. The surface of the printed objects was modified through imidazolium-based supported ionic liquid phases. Immobilisation conditions for an enzymatic solution with alcohol dehydrogenase (ADH-200) were rapidly evaluated in a 96-well format using a colorimetric assay that enables quantitative comparison of activity and loading across multiples of conditions in a parallel fashion. Methyl-imidazolium-modified supports showed markedly higher enzymatic activity and immobilisation efficiency than other imidazolium-based ionic liquids, and the best-performing formulation was directly applied to functionalise the 3D-printed honeycomb-structured reactor. The resulting heterogeneous biocatalyst catalysed the oxidation of 1-phenylethanol to acetophenone under continuous flow with high conversion at moderate residence times and sustained performance over several hundred hours on stream. This integrated 3D-printed platform facilitates the identification of effective immobilisation chemistries and their translation to robust flow reactors, providing a general strategy for the rapid development of immobilised biocatalysts.

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Journal
ChemSusChem
Published
2026-06-18
DOI
https://doi.org/10.1002/cssc.70815
Primary Topic
Innovative Microfluidic and Catalytic Techniques Innovation
Type
article
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article

3D‐Printed Platforms for Enzyme Immobilisation Screening and Direct Translation into Continuous‐Flow Biocatalysis

Paulo Roberto de Oliveira, Marcileia Zanatta, Víctor Sans, Cristopher Tinajero et al.
ChemSusChem
Innovative Microfluidic and Catalytic Techniques Innovation
article

3D‐Printed Platforms for Enzyme Immobilisation Screening and Direct Translation into Continuous‐Flow Biocatalysis

Paulo Roberto de Oliveira, Marcileia Zanatta, Víctor Sans, Cristopher Tinajero, Gianluca Palmara, Simone Marchetti, Luis David Lisintuña
article en

Abstract

The development of immobilised biocatalysts for continuous-flow processes is still largely based on laborious trial-and-error screening, and conditions optimised on conventional carriers are often not directly transferable to structured reactors. Here, we introduce a 3D-printed methodology that integrates high-throughput screening of enzyme immobilisation with implementation in a continuous-flow reactor using the same photopolymeric formulation. The surface of the printed objects was modified through imidazolium-based supported ionic liquid phases. Immobilisation conditions for an enzymatic solution with alcohol dehydrogenase (ADH-200) were rapidly evaluated in a 96-well format using a colorimetric assay that enables quantitative comparison of activity and loading across multiples of conditions in a parallel fashion. Methyl-imidazolium-modified supports showed markedly higher enzymatic activity and immobilisation efficiency than other imidazolium-based ionic liquids, and the best-performing formulation was directly applied to functionalise the 3D-printed honeycomb-structured reactor. The resulting heterogeneous biocatalyst catalysed the oxidation of 1-phenylethanol to acetophenone under continuous flow with high conversion at moderate residence times and sustained performance over several hundred hours on stream. This integrated 3D-printed platform facilitates the identification of effective immobilisation chemistries and their translation to robust flow reactors, providing a general strategy for the rapid development of immobilised biocatalysts.

ChemSusChemVol. 19(12)
Universitat Jaume I (ES)
Openalex Percentile: Top 12%
Innovative Microfluidic and Catalytic Techniques Innovation
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