Optimized Spatial Configuration of Heterogeneous BiocatalystsMaximizes Cell-Free Biosynthesis of ¿-Hydroxy and ¿-Amino Acids

Santiago-Arcos, Javier ; Velasco-Lozano, Susana ; Diamanti, Eleftheria ; Benítez-Mateos, Ana I. ; Grajales-Hernández, Daniel ; Paradisi, Francesca ; López-Gallego, Fernando

Resumen: Cell-free biocatalysis is gaining momentum in producing value-added chemicals, particularly in stepwise reaction cascades. However, the stability of enzyme cascades in industrial settings is often compromised when free enzymes are involved. In this study, we have developed a stable multifunctional heterogeneous biocatalyst coimmobilizing five enzymes on microparticles to transform 1,ω-diols into 1,ω-hydroxy acids. We improved the operational efficiency and stability of the heterogeneous biocatalyst by fine-tuning the enzyme loading and spatial organization. Stability issues are overcome through postimmobilization polymer coating. The general applicability of this heterogeneous biocatalyst is demonstrated by its scale-up in both batch and packed bed reactors, allowing a product yield of >80%. The continuous process is fed with H2O2 as the oxygen source, reaching a space-time yield (STY) of 0.76 g·L–1·h–1, maintained for the first 12 h. Finally, this flow system is telescoped with a second plug-flow reactor packed with a different heterogeneous biocatalyst integrating an additional transaminase. As a result, this 6-enzyme 2-reactor system sequentially transforms 1,ω-diols into 1,ω-amino acids while in situ recycling NAD+, depleting H2O2, and generating O2.

Idioma: Inglés

DOI: 10.1021/acssuschemeng.4c02396

Año: 2024

Publicado en: ACS Sustainable Chemistry and Engineering 12, 25 (2024), 9474-9489
ISSN: 2168-0485

Financiación: info:eu-repo/grantAgreement/ES/AEI/PRE2019-090835

Tipo y forma: Article (PostPrint)