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Enzymatic characterization of a thermostable phosphatase from Thermomicrobium roseum and its application for biosynthesis of fructose from maltodextrin

Author:
Meng, Dongdong, Liang, Ailing, Wei, Xinlei, You, Chun
Source:
Applied microbiology and biotechnology 2019 v.103 no.15 pp. 6129-6139
ISSN:
0175-7598
Subject:
Thermomicrobium roseum, biosynthesis, catalytic activity, dephosphorylation, fructose, fructose 6-phosphate, glucose, glucose 6-phosphate, glucose-6-phosphate isomerase, half life, isomerization, maltodextrins, open reading frames, pH, phosphates, phosphoglucomutase, phosphorylase, ribose, ribulose, thermal stability, thermostable enzymes
Abstract:
Phosphatases, which catalyze the dephosphorylation of compounds containing phosphate groups, are important members of the haloacid dehalogenase (HAD)–like superfamily. Herein, a thermostable phosphatase encoded by an open reading frame of Trd_1070 from Thermomicrobium roseum was enzymologically characterized. This phosphatase showed promiscuous activity against more than ten sugar phosphates, with high specific activity toward ribose 5-phosphate, followed by ribulose 5-phosphate and fructose 6-phosphate. The half-life of Trd_1070 at 70 °C and pH 7.0 was about 14.2 h. Given that the catalytic efficiency of Trd_1070 on fructose 6-phosphate was 49-fold higher than that on glucose 6-phosphate, an in vitro synthetic biosystem containing alpha-glucan phosphorylase, phosphoglucomutase, phosphoglucose isomerase, and Trd_1070 was constructed for the production of fructose from maltodextrin by whole-cell catalysis, resulting in 21.6 g/L fructose with a ratio of fructose to glucose of approximately 2:1 from 50 g/L maltodextrin. This in vitro biosystem provides an alternative method to produce fructose with higher fructose content compared with the traditional production method using glucose isomerization. Further discovery and enzymologic characterization of phosphatases may promote further production of alternative monosaccharides through in vitro synthetic biosystems.
Agid:
6504073