147103 20260217205521.0 doi 10.1007/s00253-024-13314-z sideral 140904 ART-2024-140904 eng Cinca-Fernando, Paula Universidad de Zaragoza Discovery, characterization, and synthetic potential of two novel bacterial aryl-alcohol oxidases 2024 Access copy available to the general public Unrestricted 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 info:eu-repo/grantAgreement/ES/MICINN/PID2022-136369NB-I00 info:eu-repo/grantAgreement/ES/MICINN/RYC2021-032021-I info:eu-repo/grantAgreement/EUR/MICINN/TED2021-130803B-I00 info:eu-repo/semantics/openAccess by https://creativecommons.org/licenses/by/4.0/deed.es 4.3 2024 BIOTECHNOLOGY & APPLIED MICROBIOLOGY 42 / 177 = 0.237 2024 Q1 T1 0.967 2024 Applied Microbiology and Biotechnology 2024 Q1 Medicine (miscellaneous) 2024 Q1 Biotechnology 2024 Q2 8.5 2024 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Ascaso-Alegre, Christian Universidad de Zaragoza Sevilla, Emma Universidad de Zaragoza (orcid)0000-0001-6435-3540 Martínez-Júlvez, Marta Universidad de Zaragoza (orcid)0000-0001-9047-0046 Mangas-Sánchez, Juan Ferreira, Patricia Universidad de Zaragoza (orcid)0000-0003-4076-6118 1002 412 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Fisiología Vegetal 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 108, 1 (2024) Appl. microbiol. biotechnol. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 0175-7598 1924609 http://zaguan.unizar.es/record/147103/files/texto_completo.pdf Versión publicada 2264516 http://zaguan.unizar.es/record/147103/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:147103 articulos driver 2026-02-17-20:28:43 ARTICLE