Expanding the enzymatic toolbox for β-amino acids and unnatural amino acids manufacturing

Le résumé fourni par la source

β-Amino acids, characterized by the separation of their terminal carboxylic acid and amino groups by two carbon atoms (Cα and Cβ), exhibit remarkable structural versatility. This arrangement allows for R or S isomers at both Cα and Cβ positions, resulting in up to four diastereomers for a given side chain. Such diversity facilitates the generation of a broad array of stereo- and regio-isomers, alongside the potential for di-substitution, making β-amino acids highly valuable in molecular design. Their incorporation into peptidomimetics has led to compounds with potent biological activity and enhanced resistance to proteolysis. Furthermore, β-amino acids and their derivatives serve as essential chiral building blocks in pharmaceutical synthesis, highlighting their significance in drug discovery and development. This study focuses on enzymatic approaches to β-amino acid production as a sustainable alternative to traditional chemical synthesis, which often suffers from low carbon efficiency. Leveraging both wildtype and engineered variants of aspartase and 3-methylaspartate ammonia-lyase (MAL), this project explores their catalytic potential for industrial applications. Aspartase catalyzes the reversible deamination of L-aspartic acid via a carbanion mechanism to produce fumaric acid and ammonium ion, while MAL facilitates the α,β-elimination of ammonia from 3-methylaspartate to form mesaconate. Novel aspartase and MAL enzymes sourced from thermophilic organisms were identified and characterized. Protocols for their recombinant expression and purification were established, and their enzymatic activities were validated through NMR and spectrometric assays. This research provides critical insights into the catalytic mechanisms and industrial applicability of aspartase and MAL, offering a foundation for environmentally friendly and efficient production of β-amino acids and other unnatural amino acids.

Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.