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Engineering the Donor Selectivity of D ‐Fructose‐6‐Phosphate Aldolase for Biocatalytic Asymmetric Cross‐Aldol Additions of Glycolaldehyde
Author(s) -
Szekrenyi Anna,
Soler Anna,
Garrabou Xavier,
GuérardHélaine Christine,
Parella Teodor,
Joglar Jesús,
Lemaire Marielle,
Bujons Jordi,
Clapés Pere
Publication year - 2014
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201403281
Subject(s) - glycolaldehyde , aldol reaction , chemistry , glyceraldehyde , aldolase a , saturated mutagenesis , selectivity , adduct , dihydroxyacetone , stereochemistry , organic chemistry , combinatorial chemistry , catalysis , biochemistry , enzyme , mutant , glycerol , dehydrogenase , gene
Abstract D ‐Fructose‐6‐phosphate aldolase (FSA) is a unique catalyst for asymmetric cross‐aldol additions of glycolaldehyde. A combination of a structure‐guided approach of saturation mutagenesis, site‐directed mutagenesis, and computational modeling was applied to construct a set of FSA variants that improved the catalytic efficiency towards glycolaldehyde dimerization up to 1800‐fold. A combination of mutations in positions L107, A129, and A165 provided a toolbox of FSA variants that expand the synthetic possibilities towards the preparation of aldose‐like carbohydrate compounds. The new FSA variants were applied as highly efficient catalysts for cross‐aldol additions of glycolaldehyde to N ‐carbobenzyloxyaminoaldehydes to furnish between 80–98 % aldol adduct under optimized reaction conditions. Donor competition experiments showed high selectivity for glycolaldehyde relative to dihydroxyacetone or hydroxyacetone. These results demonstrate the exceptional malleability of the active site in FSA, which can be remodeled to accept a wide spectrum of donor and acceptor substrates with high efficiency and selectivity.