biostudies-literatureAshley BRCUK | Biotechnology and Biological Sciences Research CouncilBiotechnology and Biological Sciences Research Council78https://www.ebi.ac.uk/biostudies/studies/S-EPMC11906848biostudies-literatureUnknown8(1)Carbon-carbon bond formation is one of the key pillars of organic synthesis. Green, selective and efficient biocatalytic methods for such are therefore highly desirable. The α-oxoamine synthases (AOSs) are a class of pyridoxal 5'-phosphate (PLP)-dependent, irreversible, carbon-carbon bond-forming enzymes, which have been limited previously by their narrow substrate specificity and requirement of acyl-CoA thioester substrates. We recently characterized a thermophilic enzyme from Thermus thermophilus (ThAOS) with a much broader substrate scope and described its use in a chemo-biocatalytic cascade process to generate pyrroles in good yields and timescales. Herein, we report the structure-guided engineering of ThAOS to arrive at variants able to use a greatly expanded range of amino acid and sCommunications chemistryRational engineering of a thermostable α-oxoamine synthase biocatalyst expands the substrate scope and synthetic applicability.PMC11906848BB/J01446X/1BB/T016841/1Zhu YBasle AMarles-Wright JCampopiano DJNovikovs NSajjad MAshley BMathew SfalseRational engineering of a thermostable α-oxoamine synthase biocatalyst expands the substrate scope and synthetic applicability.Carbon-carbon bond formation is one of the key pillars of organic synthesis. Green, selective and efficient biocatalytic methods for such are therefore highly desirable. The α-oxoamine synthases (AOSs) are a class of pyridoxal 5'-phosphate (PLP)-dependent, irreversible, carbon-carbon bond-forming enzymes, which have been limited previously by their narrow substrate specificity and requirement of acyl-CoA thioester substrates. We recently characterized a thermophilic enzyme from Thermus thermophilus (ThAOS) with a much broader substrate scope and described its use in a chemo-biocatalytic cascade process to generate pyrroles in good yields and timescales. Herein, we report the structure-guided engineering of ThAOS to arrive at variants able to use a greatly expanded range of amino acid and s2025-01-01T00:00:00Z2025 Mar2026-04-08T19:53:32.315Z2026-04-08T14:31:25.857ZS-EPMC119068484008270510.1038/s42004-025-01448-8