Discovery, characterization, and synthetic potential of two novel bacterial aryl-alcohol oxidases

Cinca-Fernando, Paula (Universidad de Zaragoza) ; Ascaso-Alegre, Christian (Universidad de Zaragoza) ; Sevilla, Emma (Universidad de Zaragoza) ; Martínez-Júlvez, Marta (Universidad de Zaragoza) ; Mangas-Sánchez, Juan ; Ferreira, Patricia (Universidad de Zaragoza)

Resumen: The search for novel synthetic tools to prepare industrial chemicals in a safer and greener manner is a continuing challenge in synthetic chemistry. In this manuscript, we report the discovery, characterization, and synthetic potential of two novel aryl-alcohol oxidases from bacteria which are able to oxidize a variety of aliphatic and aromatic alcohols with efficiencies up to 4970 min−1 mM−1. Both enzymes have shown a reasonable thermostability (thermal melting temperature values of 50.9 and 48.6 °C for ShAAO and SdAAO, respectively). Crystal structures revealed an unusual wide-open entrance to the active-site pockets compared to that previously described for traditional fungal aryl-alcohol oxidases, which could be associated with differences observed in substrate scope, catalytic efficiency, and other functional properties. Preparative-scale reactions and the ability to operate at high substrate loadings also demonstrate the potential of these enzymes in synthetic chemistry with total turnover numbers > 38000. Moreover, their availability as soluble and active recombinant proteins enabled their use as cell-free extracts which further highlights their potential for the large-scale production of carbonyl compounds.
Key points
• Identification and characterization of two novel bacterial aryl-alcohol oxidases
• Crystal structures reveal wide-open active-site pockets, impacting substrate scope
• Total turnover numbers and cell-free extracts demonstrate the synthetic potential

Idioma: Inglés

DOI: 10.1007/s00253-024-13314-z

Año: 2024

Publicado en: APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 108, 1 (2024)
ISSN: 0175-7598

Financiación: info:eu-repo/grantAgreement/ES/MICINN/PID2022-136369NB-I00
Financiación: info:eu-repo/grantAgreement/ES/MICINN/RYC2021-032021-I
Financiación: info:eu-repo/grantAgreement/EUR/MICINN/TED2021-130803B-I00

Tipo y forma: Artículo (Versión definitiva)

Área (Departamento): Área Fisiología Vegetal (Dpto. Bioq.Biolog.Mol. Celular)
Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)