Engineering a Thermostable Transketolase for Unnatural Conversion of (2S)-Hydroxyaldehyde. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Advanced Synthesis and Catalysis Année : 2015

Engineering a Thermostable Transketolase for Unnatural Conversion of (2S)-Hydroxyaldehyde.

Résumé

Transketolase (TK) from various origins (including Escherichia coli and yeast) has been described to be fully enantiomer specific for (2R)-hydroxyaldehyde substrates. A thermostable TK from Geobacillus stearothermophilus (TKgst) was found to display a minor reactivity for (2S)-hydroxylated aldehydes. To improve this activity by directed protein evolution, we have built a library of TKgst variants by site saturation mutagenesis on two key positions L382 and D470. The best TKgst double mutant L382D/D470S shows up to 4- and 5-fold higher activities towards L-lactaldehyde and L-glyceraldehyde as acceptor substrates, respectively. Preparative utility of this mutant was demonstrated by the one-step synthesis of valuable L-ribulose and its 5-deoxy analogue with the L-erythro (3S,4S) configuration, which were previously inaccessible by using common TK sources.

Domaines

Chimie

Dates et versions

hal-01206477 , version 1 (29-09-2015)

Identifiants

Citer

Juliane Abdoul-Zabar, Marion Lorillière, Dong Yi, Thangavelu Saravanan, Titu Devamani, et al.. Engineering a Thermostable Transketolase for Unnatural Conversion of (2S)-Hydroxyaldehyde.. Advanced Synthesis and Catalysis, 2015, 357, pp.1715-1720. ⟨10.1002/adsc.201500207⟩. ⟨hal-01206477⟩
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