α-kétoglutarate dependante Dioxygenases  (α-KetoAcid Oxygenases, αKAO)

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    • PI: Anne Zaparucha, Pierre-Loïc Saïdi

    Oxidation reactions involving non-activated C-H bonds are among the most difficult to carry out in chemistry, but they are also among the most fundamental. We have explored the catalytic capacities of new KAO, a family of enzymes that catalyse the hydroxylation reaction as an alternative to cytochrome P450.

     

     

    We have identified αKAO active on lysine with regioselectivity at C3 or C4 (KDOs). This complementarity enables the synthesis of dihydroxylated lysine by cascade reaction.1
    A structural and computational analysis of the active sites based on the resolution of the structures of two of the enzymes discovered (collaboration with T. Isabet and P. Legrand, SOLEIL synchrotron) enabled us to identify the structural elements responsible for the substrate specificity and regioselectivity of the hydroxylation reaction of amino acids and derivatives.2,3

     

     

    Various hydroxylated polyamines have been synthesised from L-lysine by linking one or two regiodivergent hydroxylation steps with decarboxylation. This very short synthesis provides access to chiral platform molecules.4

     

     

    1. Baud D., Saaidi P.-L., Monfleur A., Harari M., Cuccaro J., Fossey A., Besnard M., Debard A., Mariage A., Pellouin V., Petit J.-L., Salanoubat M., Weissenbach J., de Berardinis V., Zaparucha A., Synthesis of Mono- and Dihydroxylated Amino Acids with New α-Ketoglutarate-Dependent Dioxygenases: Biocatalytic Oxidation of CH Bonds; Chem. Cat. Chem. 2014, 6, 3012-3017; DOI:10.1002/cctc.201402498
    2. Bastard K., Isabet T., Stura E. A., Legrand P., Zaparucha A.; Structural Studies based on two Lysine Dioxygenases with Distinct Regioselectivity Brings Insights Into Enzyme Specificity within the Clavaminate Synthase-Like Family; Sci. Rep. 2018, 8:16587; DOI:10.1038/s41598-018-34795-9
    3. A. Zaparucha, V. de Berardinis, P.-L. Saaidi, D. Baud, WO2015078889
    4. Baud D., Peruch O., Saaidi P.-L., Fossey A., Mariage A., Petit J.-L., Salanoubat M., Vergne-Vaxelaire C., de Berardinis V., Zaparucha A., Biocatalytic Approaches towards the Synthesis of Chiral Amino Alcohols from Lysine: Cascade Reactions Combining AlphaKetoAcid Oxygenase Hydroxylation with Pyridoxal Phosphate Dependent Decarboxylation; Adv. Syn. Catal. 2017, 359, 1563-1569; DOI:10.1002/adsc.201600934
    5. Fossey-Jouenne A., Vergne-Vaxelaire C., Zaparucha A., Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine; J. Vis. Exp. 2018, 132, e56926; DOI:10.3791/56926.

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