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February 24, 2026The Plant Journal0 citations

Exploring the substrate promiscuity and functional residues of UGT73 family enzymes in Entada phaseoloides

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XXXun XuRTRuiheng TangLHLin He

Key Points

  • This research aims to investigate the substrate promiscuity and functional residues of the UGT73 enzyme family in Entada phaseoloides.
  • Conducted phylogenetic analysis across multiple species to observe UGT73 family expansions.
  • Re-annotated the E. phaseoloides genome using Oxford Nanopore Technologies and Illumina data.
  • Selected and characterized four UGT73 family genes for functional analysis.
  • Used structural modeling and molecular docking to identify key active sites.
  • Employed site-directed mutagenesis to test critical residues affecting activity.
  • Identified UGT73AA6 as specific for flavonoids, while UGT73CG48 and UGT73CG49 also glycosylate triterpenoids.
  • UGT73CG49 exhibited higher activity for glucosylation over xylosylation in both flavonoids and triterpenes.
  • Gly194 was revealed as a critical residue that enhances catalytic activity in UGT73CG49.

Abstract

SUMMARY Flavonoid glycosides and triterpenoid saponins are bioactive plant metabolites with broad applications in food, medicine, and agriculture. These compounds are typically synthesized through glycosylation catalyzed by uridine diphosphate‐dependent glycosyltransferases (UGTs). In this study, phylogenetic analysis across multiple species revealed a lineage‐specific expansion of the UGT73 family in legumes such as Entada phaseoloides and Glycine max . The genome of the medicinal legume E. phaseoloides was re‐annotated using integrated Oxford Nanopore Technologies and Illumina transcriptomic data to identify target genes. Four expanded UGT73 family genes were selected and functionally characterized. UGT73AA6 specifically glycosylates flavonoids, while UGT73CG48 and UGT73CG49 catalyze glycosylation of both flavonoids and pentacyclic triterpenoids. UGT73CG49 exhibits higher catalytic activity for the glucosylation of flavonoids and pentacyclic triterpenes compared to its xylosylation activity. Structural modeling and molecular docking identified key active sites, and site‐directed mutagenesis revealed Gly194 as a critical residue enhancing catalytic activity in UGT73CG49. This study provides new insights into the functional evolution and metabolic versatility of the UGT73 family in legumes. The identification and engineering of UGT73 genes from E. phaseoloides lay a foundation for future applications in biosynthetic pathway engineering and the industrial production of high‐value glycosides.

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Cite This Study

Xu et al. (2026) studied this question.

synapsesocial.com/papers/699d3ff8de8e28729cf64ed1https://doi.org/10.1111/tpj.70739
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