Structure-based prediction and identification of 4-epimerization activity of phosphate sugars in class II aldolases

Scientific Reports
Seon-Hwa LeeDeok-Kun Oh

Abstract

Sugar 4-epimerization reactions are important for the production of rare sugars and their derivatives, which have various potential industrial applications. For example, the production of tagatose, a functional sweetener, from fructose by sugar 4-epimerization is currently constrained because a fructose 4-epimerase does not exist in nature. We found that class II D-fructose-1,6-bisphosphate aldolase (FbaA) catalyzed the 4-epimerization of D-fructose-6-phosphate (F6P) to D-tagatose-6-phosphate (T6P) based on the prediction via structural comparisons with epimerase and molecular docking and the identification of the condensed products of C3 sugars. In vivo, the 4-epimerization activity of FbaA is normally repressed. This can be explained by our results showing the catalytic efficiency of D-fructose-6-phosphate kinase for F6P phosphorylation was significantly higher than that of FbaA for F6P epimerization. Here, we identified the epimerization reactions and the responsible catalytic residues through observation of the reactions of FbaA and L-rhamnulose-1-phosphate aldolases (RhaD) variants with substituted catalytic residues using different substrates. Moreover, we obtained detailed potential epimerization reaction mechanism of Fb...Continue Reading

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Citations

Mar 30, 2018·The Journal of Biological Chemistry·Benoit JacquesJurgen Sygusch
Oct 12, 2018·Microbial Biotechnology·Agnieszka SekowskaAntoine Danchin

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Methods Mentioned

BETA
circular dichroism
PCR
enzymatic assay
NMR

Software Mentioned

FATCAT
Discovery Studio
Clustal W2
DOCKER
Twists FATCAT
CHARMM
CDOCKER
DS

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