TY - JOUR
T1 - Structure-guided engineering of D-fructose-6-phosphate aldolase for improved acceptor tolerance in biocatalytic aldol additions
AU - Soler, Anna
AU - Gutiérrez, Mariana L.
AU - Bujons, Jordi
AU - Parella, Teodor
AU - Minguillon, Cristina
AU - Joglar, Jesús
AU - Clapés, Pere
PY - 2015/5/1
Y1 - 2015/5/1
N2 - © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Abstract A structure-guided redesign of D-fructose-6-phosphate aldolase from Escherichia coli (FSA) was devised for improving the acceptor tolerance towards α-substituted and conformationally constrained aldehydes. FSA A129S/R134X/A165G/S166G and L107Y/A129G/R134X/A165G/S166G variants, where X was R, V, P, or S, were the most suited biocatalysts for dihydroxyacetone, hydroxyacetone and glycolaldehyde additions to 20 α-substituted N-Cbz-aminoaldehydes (Cbz=benzyloxycarbonyl) including pyrrolidine and piperidine derivatives. Full kinetic stereocontrol for si-si face addition of the aldolase-bound nucleophile to the N-Cbz-aminoaldehyde carbonyl was observed, furnishing the corresponding D-threo configured aldol adduct in >95:5 dr as assessed by NMR. After reductive amination, 47 different iminocyclitols were identified and characterized. In some examples partial racemization of the corresponding aldehyde was observed, which appears to be produced mostly during the aldol addition reactions.
AB - © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Abstract A structure-guided redesign of D-fructose-6-phosphate aldolase from Escherichia coli (FSA) was devised for improving the acceptor tolerance towards α-substituted and conformationally constrained aldehydes. FSA A129S/R134X/A165G/S166G and L107Y/A129G/R134X/A165G/S166G variants, where X was R, V, P, or S, were the most suited biocatalysts for dihydroxyacetone, hydroxyacetone and glycolaldehyde additions to 20 α-substituted N-Cbz-aminoaldehydes (Cbz=benzyloxycarbonyl) including pyrrolidine and piperidine derivatives. Full kinetic stereocontrol for si-si face addition of the aldolase-bound nucleophile to the N-Cbz-aminoaldehyde carbonyl was observed, furnishing the corresponding D-threo configured aldol adduct in >95:5 dr as assessed by NMR. After reductive amination, 47 different iminocyclitols were identified and characterized. In some examples partial racemization of the corresponding aldehyde was observed, which appears to be produced mostly during the aldol addition reactions.
KW - aldol reaction
KW - aldolases
KW - D -fructose-6-phosphate aldolase
KW - enzyme catalysis
KW - iminocyclitols
KW - lyases
KW - mutagenesis
UR - https://www.scopus.com/pages/publications/85027924338
U2 - 10.1002/adsc.201500073
DO - 10.1002/adsc.201500073
M3 - Article
SN - 1615-4150
VL - 357
SP - 1787
EP - 1807
JO - Advanced Synthesis and Catalysis
JF - Advanced Synthesis and Catalysis
IS - 8
ER -