Structure-guided engineering of D-fructose-6-phosphate aldolase for improved acceptor tolerance in biocatalytic aldol additions

Anna Soler, Mariana L. Gutiérrez, Jordi Bujons, Teodor Parella, Cristina Minguillon, Jesús Joglar, Pere Clapés

    Research output: Contribution to journalArticleResearchpeer-review

    14 Citations (Scopus)

    Abstract

    © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Abstract A structure-guided redesign of D-fructose-6-phosphate aldolase from Escherichia coli (FSA) was devised for improving the acceptor tolerance towards α-substituted and conformationally constrained aldehydes. FSA A129S/R134X/A165G/S166G and L107Y/A129G/R134X/A165G/S166G variants, where X was R, V, P, or S, were the most suited biocatalysts for dihydroxyacetone, hydroxyacetone and glycolaldehyde additions to 20 α-substituted N-Cbz-aminoaldehydes (Cbz=benzyloxycarbonyl) including pyrrolidine and piperidine derivatives. Full kinetic stereocontrol for si-si face addition of the aldolase-bound nucleophile to the N-Cbz-aminoaldehyde carbonyl was observed, furnishing the corresponding D-threo configured aldol adduct in >95:5 dr as assessed by NMR. After reductive amination, 47 different iminocyclitols were identified and characterized. In some examples partial racemization of the corresponding aldehyde was observed, which appears to be produced mostly during the aldol addition reactions.
    Original languageEnglish
    Pages (from-to)1787-1807
    JournalAdvanced Synthesis and Catalysis
    Volume357
    Issue number8
    DOIs
    Publication statusPublished - 1 May 2015

    Keywords

    • aldol reaction
    • aldolases
    • D -fructose-6-phosphate aldolase
    • enzyme catalysis
    • iminocyclitols
    • lyases
    • mutagenesis

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