Abstract
Objective
To produce tagatose from fructose with a high conversion rate and to establish a high-yield purification method of tagatose from the reaction mixture.
Results
Fructose at 1 M (180 g l−1) was converted to 0.8 M (144 g l−1) tagatose by a three-step enzymatic cascade reaction, involving hexokinase, plus ATP, fructose-1,6-biphosphate aldolase, phytase, over 16 h with a productivity of 9 g l−1 h−1. No byproducts were detected. Tagatose was recrystallized from ethanol to a purity of 99.9% and a yield of 96.3%. Overall, tagatose at 99.9% purity was obtained from fructose with a yield of 77%.
Conclusion
This is the first biotechnological production of tagatose from fructose and the first application of solvent recrystallization for the purification of rare sugars.
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Acknowledgements
This study was supported by the Ministry of Trade, Industry & Energy (MOTIE), Korea Institute for Advancement of Technology (KIAT) through the Encouragement Program for The Industries of Economic Cooperation Region.
Supporting information
Supplementary Fig. 1—Bio-LC profiles of (I) substrate and (II) product in the conversion of F6P to T6P by FbaA.
Supplementary Fig. 2—Effects of the pH and temperature of FbaA on the 4-epimerization of F6P to T6P by FbaA.
Supplementary Fig. 3—Metabolic pathway engineering for the overproduction of tagatose from glucose or fructose and glycerol in E. coli.
Supplementary Fig. 4—Purification of tagatose from the reaction mixture with fructose by ion chromatography using a column containing Amberite CR 1310 (Ca2+ form) resin.
Supplementary Fig. 5—HPLC profiles during the separation of tagatose from the reaction mixture with fructose by recrystallization from ethanol.
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Lee, SH., Hong, SH., Kim, KR. et al. High-yield production of pure tagatose from fructose by a three-step enzymatic cascade reaction. Biotechnol Lett 39, 1141–1148 (2017). https://doi.org/10.1007/s10529-017-2340-3
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DOI: https://doi.org/10.1007/s10529-017-2340-3