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Engineering a branching sucrase for flavonoid glucoside diversification.

Yannick Malbert1, Claire Moulis1, Yoann Brison1

  • 1Laboratoire d'Ingénierie des Systèmes Biologiques et Procédés, LISBP, Université de Toulouse, CNRS, INRA, INSA, Toulouse, France. 135, avenue de Rangueil, F-31077, Toulouse, cedex 04, France.

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Engineered enzymes efficiently create novel flavonoid glycosides, enhancing solubility and bioavailability for health applications. This study advances enzymatic modification of flavonoids, including previously difficult compounds.

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Area of Science:

  • Biocatalysis
  • Enzymology
  • Glycochemistry

Background:

  • Enzymatic glycosylation of flavonoids enhances their properties like solubility, stability, and bioavailability, crucial for healthcare applications.
  • Developing efficient enzymatic platforms for flavonoid modification is essential for expanding their therapeutic potential.

Purpose of the Study:

  • To engineer an α-transglucosylase (glycoside-hydrolase family 70) for efficient flavonoid glucosylation.
  • To create a versatile enzymatic platform for producing diverse flavonoid glycosides.

Main Methods:

  • Engineering of an α-1,2 branching sucrase (GH70) through targeted mutations guided by molecular docking.
  • Screening of enzyme variants for quercetin glucosylation using sucrose as a glucosyl donor.
  • Assessing the acceptor promiscuity of selected variants towards other flavonoids.

Main Results:

  • Identified 22 promising enzyme variants with improved quercetin conversion and product profiles.
  • Demonstrated significant glucosylation of luteolin, morin, and naringenin (30-90% conversion) using engineered variants.
  • Successfully glucosylated naringenin and morin, which are typically recalcitrant to α-transglucosylases.

Conclusions:

  • Engineered GH70 α-transglucosylases offer a powerful platform for flavonoid glycosylation.
  • This approach expands the accessible chemical diversity of flavonoid glycosides, including previously challenging structures.
  • The study opens new avenues for producing novel flavonoid derivatives with enhanced bioavailability for health applications.