Enzymatic Synthesis of Aliphatic Acyloins through Thermostable Transketolase Catalyzed-Reaction
Résumé
Thermostable TKgst was successfully engineered for the synthesis of aliphatic acyloins with various carbon backbone lengths (C5-C10). Based on structure guided studies, efficient TKgst variants with enhanced activities toward aliphatic aldehydes as acceptors together with aliphatic pyruvate homologues as donors were identified. The TKgst single variant L382F was able to catalyze efficiently the transfer of the ketol group from HPA on all targeted aliphatic aldehydes (C3-C8) to give the corresponding 1,3-dihydroxy ketones with good yields and excellent enantioselectivity. The combination of H102L/H474S previously designed for the improvement toward aliphatic pyruvate homologues, with F435I gave a new variant H102L/H474S/F435I able to transfer the acyl goup of 2-oxobutyrate and 2-oxovalerate to aliphatic aldehydes, giving mono hydroxylated ketones never obtained before with this enzyme.
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