Engineering the Donor Selectivity of D-Fructose-6-Phosphate Aldolase for Biocatalytic Asymmetric Cross-Aldol Additions of Glycolaldehyde

Anna Szekrenyi, Anna Soler, Xavier Garrabou, Christine Guérard-Hélaine, Teodor Parella, Jesús Joglar, Marielle Lemaire, Jordi Bujons, Pere Clapés

    Research output: Contribution to journalArticleResearchpeer-review

    28 Citations (Scopus)

    Abstract

    © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim D-Fructose-6-phosphate aldolase (FSA) is a unique catalyst for asymmetric cross-aldol additions of glycolaldehyde. A combination of a structure-guided approach of saturation mutagenesis, site-directed mutagenesis, and computational modeling was applied to construct a set of FSA variants that improved the catalytic efficiency towards glycolaldehyde dimerization up to 1800-fold. A combination of mutations in positions L107, A129, and A165 provided a toolbox of FSA variants that expand the synthetic possibilities towards the preparation of aldose-like carbohydrate compounds. The new FSA variants were applied as highly efficient catalysts for cross-aldol additions of glycolaldehyde to N-carbobenzyloxyaminoaldehydes to furnish between 80–98 % aldol adduct under optimized reaction conditions. Donor competition experiments showed high selectivity for glycolaldehyde relative to dihydroxyacetone or hydroxyacetone. These results demonstrate the exceptional malleability of the active site in FSA, which can be remodeled to accept a wide spectrum of donor and acceptor substrates with high efficiency and selectivity.
    Original languageEnglish
    Pages (from-to)12572-12583
    JournalChemistry - A European Journal
    Volume20
    Issue number39
    DOIs
    Publication statusPublished - 1 Jan 2014

    Keywords

    • aldol reaction
    • carbohydrates
    • enzyme catalysis
    • enzyme models
    • protein engineering

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