Food and Fermentation Industries >

2021 , Vol. 47 >Issue 24: 102 - 108

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.027355

Properties and application of sucrose synthase from Nitrosospira multiformis

  • FENG Xin ,
  • ZHAO Liting ,
  • GU Zhenghua ,
  • LI Youran ,
  • SHI Guiyang ,
  • DING Zhongyang
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  • 1(Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China)
    2(National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China)
    3(School of Biotechnology, Jiangnan University, Wuxi 214122, China)

Received date: 2021-03-17

  Revised date: 2021-04-23

  Online published: 2022-01-21

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Abstract

UDP-glucose is an important sugar donor for glycosylation of “Leloir” glycosyltransferases (GTs). Sucrose synthase (SuSy) can utilize both sucrose and UDP as substrates to reversibly generate UDP-glucose in a one-step reaction. The SuSy gene from Nitrosospira multiformis(NmSuSy) was heterologously expressed in Escherichia coli. The purified NmSuSy was analyzed for its enzymatic properties and functions. The efficient synthesis of UDP-glucose was achieved by overcoming the thermodynamic and kinetic constraints of the reversible reaction. Results showed that the molecular weight of SuSy was 89 kDa, and the optimum reaction pH for SuSy was 6.5 and it was stable ranging from pH 6.0 to 7.5; while the optimal temperature was 55 ℃ and it behaved better with temperature lower than 40 ℃. Under the optimized conditions (pH 6, temperature 40 ℃, 1 mol/L sucrose, 100 mmol/L UDP), reactions with NmSuSy (100 mg/L) could produce 35.8 mmol/L (20.2 g/L) of UDP-glucose in 1 h. When the reversible reaction reached equilibrium within 6 hours, 41.5 mmol/L (23.5 g/L) of UDP-glucose could be produced. This study lays a theoretical foundation for the efficient synthesis of UDP-glucose, and provides a more economical donor substrate material for the research of glycobiology.

Key words： Nitrosospira multiformis; sucrose synthase; UDP-glucose; heterologous expression; enzymatic properties

Cite this article

FENG Xin , ZHAO Liting , GU Zhenghua , LI Youran , SHI Guiyang , DING Zhongyang . Properties and application of sucrose synthase from Nitrosospira multiformis[J]. Food and Fermentation Industries, 2021 , 47(24) : 102 -108 . DOI: 10.13995/j.cnki.11-1802/ts.027355

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