TY - JOUR
T1 - Engineering a thermostable transketolase for unnatural conversion of (2S)-hydroxyaldehydes
AU - Abdoul Zabar, Juliane
AU - Lorillière, Marion
AU - Yi, Dong
AU - Thangavelu, Saravanan
AU - Devamani, Titu
AU - Nauton, Lionel
AU - Charmantray, Franck
AU - Hélaine, Virgil
AU - Fessner, Wolf Dieter
AU - Hecquet, Laurence
N1 - Publisher Copyright:
© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
PY - 2015/5/1
Y1 - 2015/5/1
N2 - 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.
AB - 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.
KW - carboligation
KW - mutagenesis
KW - stereoselectivity
KW - transketolase
KW - α-hydroxylated aldehydes
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U2 - 10.1002/adsc.201500207
DO - 10.1002/adsc.201500207
M3 - Article
AN - SCOPUS:85027953300
SN - 1615-4150
VL - 357
SP - 1715
EP - 1720
JO - Advanced Synthesis and Catalysis
JF - Advanced Synthesis and Catalysis
IS - 8
ER -