Date: 13 June 2025 | Category: News
Authors: Javier Santiago-Arcos, Susana Velasco-Lozano, Eleftheria Diamanti, Ana I. Benítez-Mateos, Daniel Grajales-Hernández, Francesca Paradisi, and Fernando López-Gallego
Researchers at the Heterogeneous Biocatalysis Laboratory at CIC biomaGUNE, led by Drs. Fernando López Gallego and Susana Velasco-Lozano, have successfully developed a stable multifunctional heterogeneous biocatalyst that co-immobilizes five enzymes onto microparticles, transforming 1,ω-diols into 1,ω-hydroxy acids [1]. Scale-up using a packed-bed reactor (PBR) enabled yields of over 80%, and a space-time yield of 0.76 g/L/hour, which was maintained over 12 hours. Telescoping with a second PBR containing a transaminase allowed simple conversion of 1,ω-diols into 1,ω-amino acids in >95% conversion.
Cell-free biocatalysis: a useful tool for chemical manufacturing
When working with multi-step reaction cascades, cell-free biocatalysis is rapidly coming to the fore as a key enabling technology. Enzymes can be readily incorporated into synthetic routes and allow for efficient and sustainable chemical manufacturing, with the ability to undertake several transformations in one pot, avoiding numerous work-up and purification processes [2]. However, within industrial processes, free enzymes can cause instability in enzyme cascades, therefore enzyme immobilisation has emerged as a useful tool for enhancing enzyme stability and allowing enzyme recycling [3].
In this work, a 5-enzyme system was exploited to convert 1,5-pentanediol into 5-hydroxypentanoic acid, with the five immobilised enzymes being stabilised through polymeric coating post-immobilisation and placed in a PBR. Passing the crude 5-hydroxypentanoic acid solution through a second PBR containing an immobilised transaminase enabled facile conversion of the intermediate into 5-aminopentanoic acid, which has potential use within nylon synthesis [4], Figure 2. Recycling of material at a flow rate of 0.02 mL/min for 72 hours provided over 95% conversion of the diol to the corresponding aminopentanoic acid, with 3.8 mmol isolated overall.
Summary
This work shows the possibility for preparing immobilised enzymes to provide routes to complex heterogeneous biocatalytic systems, as well as the use of enabling technologies such as PBRs. Over this reaction process, six immobilised enzymes were used across two telescoped PBRs, which is a record for the number of enzymes and reaction steps assembled in a flow-mediated cell-free biosynthetic cascade. Contact us today to talk about how the use of continuous flow processing can aid your research endeavours mailto:[email protected]
References:
[1] Optimized Spatial Configuration of Heterogeneous Biocatalysts Maximizes Cell-Free Biosynthesis of ω‑Hydroxy and ω‑Amino Acids. (J. Santiago-Arcos, S. Velasco-Lozano, E. Diamanti, A. I. Benítez-Mateos, D. Grajales-Hernández, F. Paradisi, F. López-Gallego, ACS Sustainable Chem. Eng., 2024, 12, 9474 – 9489). https://doi.org/10.1021/acssuschemeng.4c02396
[2] Recent advances in artificial enzyme cascades for the production of value-added chemicals. (Z. Wang, B. Sundara Sekar, Z. Li, Bioresource Technol., 2021, 323, 124551). https://doi.org/10.1016/j.biortech.2020.124551
[3] The limits to biocatalysis: pushing the envelope. (R. A. Sheldon, D. Brady, Chem. Commun., 2018, 54, 6088 – 6104). https://doi.org/10.1039/C8CC02463D
[4] Biocatalytic Production of a Nylon 6 Precursor from Caprolactone in Continuous Flow. (M. Romero-Fernandez, C. M. Heckmann, F. Paradisi, ChemSusChem, 2022, 15, 16, e202200811). https://doi.org/10.1002/cssc.202200811
