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Improving the Transglycosylation Activity of α-Glucosidase from Xanthomonas campestris Through Semi-rational Design for the Synthesis of Ethyl Vanillin-α-Glucoside

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Abstract

The α-glucosidase (EC 3.2.1.20) XgtA produced by Xanthomonas campestris shows high α-glucosyl transfer activity toward alcoholic and phenolic hydroxyl groups. Ethyl vanillin-α-glucoside, a precursor-aroma compound with improved water solubility and thermal stability, can be synthesized through the transglycosylation of ethyl vanillin by XgtA. However, its low ethyl vanillin-α-glucoside yield and ability to hydrolyze ethyl vanillin-α-glucoside limits for industrial applications. Rational design and site-directed mutagenesis were employed to generate three variants of X. campestris α-glucosidase, L145I, S272T, and L145I/S272T, with improved transglycosylation activity toward EV. The highest yield is up to 52.41% by L145I/S272T, which also displayed remarkably lower hydrolysis activity toward the glycoside product EVG compared to XgtA. These results also showed that the mutation in sugar-binding subsite + 1 is more effective than subsite -1 for enhancing the ratio of transglycosylation/hydrolysis for the α-glucosidase XgtA.

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Funding

The financial support provided for this project was from the National Natural Science Foundation of China (Grant No. 21908196).

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Authors and Affiliations

  1. Institute of Fermentation Engineering, Zhejiang University of Technology, Hangzhou, 310014, China

    Luyi Chen, Yi Liu, Yaoyao Zhou, Linjiang Zhu & Xiaolong Chen

  2. College of Biotechnology and Bioengineering, Zhejiang University of Technology, 18 Chaowang Road, Hangzhou, 310014, China

    Luyi Chen, Yi Liu, Yaoyao Zhou, Linjiang Zhu & Xiaolong Chen

Authors
  1. Luyi Chen
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  2. Yi Liu
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  3. Yaoyao Zhou
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  4. Linjiang Zhu
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  5. Xiaolong Chen
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Contributions

LY Chen: Conceptualization, methodology, validation, formal analysis, investigation and writing original draft.

Y Liu: Investigation.

YY Zhou: Review and editing.

LJ Zhu and XL Chen: Conceptualization, funding acquisition, supervision, writing, review and editing.

Corresponding authors

Correspondence to Linjiang Zhu or Xiaolong Chen.

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Chen, L., Liu, Y., Zhou, Y. et al. Improving the Transglycosylation Activity of α-Glucosidase from Xanthomonas campestris Through Semi-rational Design for the Synthesis of Ethyl Vanillin-α-Glucoside. Appl Biochem Biotechnol 194, 3082–3096 (2022). https://doi.org/10.1007/s12010-022-03908-6

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  • DOI: https://doi.org/10.1007/s12010-022-03908-6

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