Applied Microbiology and Biotechnology

, Volume 103, Issue 23–24, pp 9423–9432 | Cite as

Insight into the glycosylation and hydrolysis kinetics of alpha-glucosidase in the synthesis of glycosides

  • Hanchi Chen
  • Shanshan Yang
  • Anjie Xu
  • Ruini Jiang
  • Zhuance Tang
  • Jiamin Wu
  • Linjiang Zhu
  • Shijie Liu
  • Xiaolong Chen
  • Yuele LuEmail author
Biotechnological products and process engineering


α-Glucosidase, Agl2, from Xanthomonas campestris was successfully overexpressed in Escherichia coli BL21(DE3) cells and purified with Ni columns. The enzyme exhibits glycosylation abilities towards a wide range of phenolic substrates, including phenol, vanillin, and ethyl vanillin, with maltose as the glycosyl donor. The catalytic properties of the purified enzyme were further investigated. It was observed that the synthesized glycosides started to degrade with prolonged catalytic time, giving an “n”-shaped kinetic profile. To understand such catalytic behavior, the Agl2-catalyzed glycosylation process was investigated kinetically. Based on the obtained parameters, it was concluded that although the substrate conversions are thermodynamically restricted in a batch system, the glycosylation efficiency can be kinetically controlled by the glycosylation/hydrolysis selectivity. Glucose was produced by both glycosylation and hydrolysis, significantly impacting the glycosylation efficiency. This study provides a mechanistic understanding of the α-glucosidase-catalyzed glycosylation process in a water-based system. The developed kinetic model was successful in explaining and analyzing the catalytic process. It is suggested that when α-glucosidase is employed for glycosylation in a water-enriched environment, the catalytic efficiency is mainly impacted by the enzyme’s glycosylation/hydrolysis selectivity and glucose content in the catalytic environment.


α-Glucosidase Glycosylation Kinetics Glycosylation/hydrolysis selectivity Glucose inhibition 


Funding information

This study was funded by the National Natural Science Foundation of China (No. 31601390, No. 21908196, and No. 21572206).

Compliance with ethical standards

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

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

253_2019_10205_MOESM1_ESM.pdf (462 kb)
ESM 1 (PDF 461 kb)


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Copyright information

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

Authors and Affiliations

  1. 1.Fermentation Technology InstituteZhejiang University of TechnologyHangzhouChina
  2. 2.Department of Paper and Bioprocess Engineering, College of Environmental Science and ForestrySUNYSyracuseUSA

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