Structural basis and mechanism of SAM-utilizing PLP-dependent enzymes

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S-adenosylmethionine (SAM)-utilizing pyridoxal phosphate (PLP)-dependent enzymes (SAM-PLP enzymes) are an emerging family of biocatalysts that stand out in terms of their remarkable chemical versatility rooted in the combined and potentiated reactivities of the carbanion-stabilizing cofactor PLP and the multifunctional nature of the sulfonium-bearing SAM. Reported enzyme-mediated transformations range from intramolecular cyclopropylations, to intermolecular Claisen-like reactions, C-N bond-forming aza-Michael-type additions, or tandem C-C bond formations via (3+2)-cycloaddition. The steadily increasing number of available genomic and metagenomic data represents a vast resource for the discovery of novel biocatalysts by targeted data mining. However, due to the current lack of in-depth structural and mechanistic knowledge for SAM-PLP enzymes, no methods are available for functional annotation based on sequence data. The goal of the project is to elucidate key structural and mechanistic determinants for SAM binding and conversion by SAM-PLP enzymes and the subsequent development of bioinformatic tools for targeted enzyme discovery approaches by genomic data mining. The results will provide a deeper understanding of the enzymological and evolutionary functions of SAM and will lay the basis to explore and exploit the versatile group of SAM-PLP biocatalysts.