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

  • Dongdong Meng
  • Ailing Liang
  • Xinlei Wei
  • Chun YouEmail author
Biotechnologically relevant enzymes and proteins


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.


Phosphatase Thermostable enzymes HAD–like hydrolase Substrate ambiguity Fructose In vitro synthetic enzymatic biosystems 


Funding information

This work was supported by the Key Research Program of the Chinese Academy of Sciences (Grant No. ZDRW-ZS-2016-3), the National Natural Science Foundation of China (Grant No. 31600635 and 21778073), and the 1000-youth talent program of China.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interest.

Ethical statement

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2019_9917_MOESM1_ESM.pdf (361 kb)
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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Dongdong Meng
    • 1
  • Ailing Liang
    • 1
    • 2
  • Xinlei Wei
    • 1
  • Chun You
    • 1
    Email author
  1. 1.Tianjin Institute of Industrial BiotechnologyChinese Academy of SciencesTianjinPeople’s Republic of China
  2. 2.School of BiotechnologyTianjin University of Science and TechnologyTianjinPeople’s Republic of China

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