biostudies-literatureAshley BDerek Stewart Charitable TrustBiotechnology and Biological Sciences Research Council7997-8002https://www.ebi.ac.uk/biostudies/studies/S-EPMC10230504biostudies-literatureUnknown11(21)We report a chemo-biocatalytic cascade for the synthesis of substituted pyrroles, driven by the action of an irreversible, thermostable, pyridoxal 5'-phosphate (PLP)-dependent, C-C bond-forming biocatalyst (<i>Th</i>AOS). The <i>Th</i>AOS catalyzes the Claisen-like condensation between various amino acids and acyl-CoA substrates to generate a range of α-aminoketones. These products are reacted with β-keto esters in an irreversible Knorr pyrrole reaction. The determination of the 1.6 Å resolution crystal structure of the PLP-bound form of <i>Th</i>AOS lays the foundation for future engineering and directed evolution. This report establishes the AOS family as useful and versatile C-C bond-forming biocatalysts.ACS sustainable chemistry & engineeringVersatile Chemo-Biocatalytic Cascade Driven by a Thermophilic and Irreversible C-C Bond-Forming α-Oxoamine Synthase.PMC10230504BB/J01446X/1BB/M010996/1BB/T016841/1BB/V00168X/1El Ashram ABasle AMarles-Wright JCampopiano DJKelis PSajjad MAshley BfalseVersatile Chemo-Biocatalytic Cascade Driven by a Thermophilic and Irreversible C-C Bond-Forming α-Oxoamine Synthase.We report a chemo-biocatalytic cascade for the synthesis of substituted pyrroles, driven by the action of an irreversible, thermostable, pyridoxal 5'-phosphate (PLP)-dependent, C-C bond-forming biocatalyst (<i>Th</i>AOS). The <i>Th</i>AOS catalyzes the Claisen-like condensation between various amino acids and acyl-CoA substrates to generate a range of α-aminoketones. These products are reacted with β-keto esters in an irreversible Knorr pyrrole reaction. The determination of the 1.6 Å resolution crystal structure of the PLP-bound form of <i>Th</i>AOS lays the foundation for future engineering and directed evolution. This report establishes the AOS family as useful and versatile C-C bond-forming biocatalysts.2023-01-01T00:00:00Z2023 May2026-05-29T10:40:03.352Z2025-04-19T15:03:08.899ZS-EPMC102305043726635410.1021/acssuschemeng.3c00243