Comparative structural analysis of plant uridine diphosphate (UDP)-dependent glycosyltransferases (UGTs) in plant specialized metabolism: structures of plant UGTs for biosynthesis of steviol glycosides

Aubuchon, Lindsey N.; Schmiederer, Kerstin; Lee, Soon Goo

doi:10.1007/s11101-023-09857-0

Cited by 4 publications

(2 citation statements)

References 93 publications

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“…UGTs active on UDP-glucose generally feature a conserved Gln-Asp/Glu-Trp-Thr amino acid tetrad, interacting with the hydroxyl groups of the donor glucose (Supplementary Fig. 33 ) 24 , 69 , 72 – 75 . To elucidate the specific donor binding interactions in the herein characterized enzymes, we employed AlphaFold for quaternary structure prediction.…”

Section: Resultsmentioning

confidence: 99%

“…Similar to most other UGTs, UGT72 enzymes form a glycosidic linkage (e.g., C−O−C, Fig. 1a ) from the phenolic hydroxyl group of the aglycone to the anomeric carbon of the sugar moiety 24 , 25 . Conversely, exclusive glucose ester-forming activities toward phenolic acids are exhibited by group L UGTs, which include UGT74, UGT75, and UGT84 family enzymes 26 .…”

Section: Introductionmentioning

confidence: 87%

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Discovery, characterization, and comparative analysis of new UGT72 and UGT84 family glycosyltransferases

Li,

Borg,

Krammer

et al. 2024

Commun Chem

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Glycosylated derivatives of natural product polyphenols display a spectrum of biological activities, rendering them critical for both nutritional and pharmacological applications. Their enzymatic synthesis by glycosyltransferases is frequently constrained by the limited repertoire of characterized enzyme-catalyzed transformations. Here, we explore the glycosylation capabilities and substrate preferences of newly identified plant uridine diphosphate (UDP)-dependent glycosyltransferases (UGTs) within the UGT72 and UGT84 families, with particular focus on natural polyphenol glycosylation from UDP-glucose. Four UGTs are classified according to their phylogenetic relationships and reaction products, identifying them as biocatalysts for either glucoside (UGT72 enzymes) or glucose ester (UGT84 members) formation from selected phenylpropanoid compounds. Detailed kinetic evaluations expose the unique attributes of these enzymes, including their specific activities and regio-selectivities towards diverse polyphenolic substrates, with product characterizations validating the capacity of UGT84 family members to perform di-O-glycosylation on flavones. Sequence analysis coupled with structural predictions through AlphaFold reveal an unexpected absence of a conserved threonine residue across all four enzymes, a trait previously linked to pentosyltransferases. This comparative analysis broadens the understood substrate specificity range for UGT72 and UGT84 enzymes, enhancing our understanding of their utility in the production of natural phenolic glycosides. The findings from this in-depth characterization provide valuable insights into the functional versatility of UGT-mediated reactions.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 87%

Discovery, characterization, and comparative analysis of new UGT72 and UGT84 family glycosyltransferases

Li,

Borg,

Krammer

et al. 2024

Commun Chem

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Molecular basis of ligand recognition specificity of flavone glucosyltransferases in Nemophila menziesii

Murayama,

Kato-Murayama,

Hosaka

et al. 2024

Archives of Biochemistry and Biophysics

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Structural insights into the catalytic selectivity of glycosyltransferase SgUGT94-289-3 towards mogrosides

Cui,

Zhang,

Wang

et al. 2024

Nat Commun

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Mogrosides constitute a series of natural sweeteners extracted from Siraitia grosvenorii fruits. These mogrosides are glucosylated to different degrees, with mogroside V (M5) and siamenoside I (SIA) being two mogrosides with high intensities of sweetness. SgUGT94-289-3 constitutes a uridine diphosphate (UDP)-dependent glycosyltransferase (UGT) responsible for the biosynthesis of M5 and SIA, by continuously catalyzing glucosylation on mogroside IIe (M2E) and on the subsequent intermediate mogroside products. However, the mechanism of its promiscuous substrate recognition and multiple catalytic modes remains unclear. Here, we report multiple complex structures and the enzymatic characterization of the glycosyltransferase SgUGT94-289-3. We show that SgUGT94-289-3 adopts a dual-pocket organization in its active site, which allows the two structurally distinct reactive ends of mogrosides to be presented from different pockets to the active site for glucosylation reaction, thus enabling both substrate promiscuity and catalytic regioselectivity. We further identified a structural motif that is essential to catalytic activity and regioselectivity, and generated SgUGT94-289-3 mutants with greatly improved M5/SIA production from M2E in an in vitro one-pot setup.

show abstract

Comparative structural analysis of plant uridine diphosphate (UDP)-dependent glycosyltransferases (UGTs) in plant specialized metabolism: structures of plant UGTs for biosynthesis of steviol glycosides

Cited by 4 publications

References 93 publications

Discovery, characterization, and comparative analysis of new UGT72 and UGT84 family glycosyltransferases

Discovery, characterization, and comparative analysis of new UGT72 and UGT84 family glycosyltransferases

Molecular basis of ligand recognition specificity of flavone glucosyltransferases in Nemophila menziesii

Structural insights into the catalytic selectivity of glycosyltransferase SgUGT94-289-3 towards mogrosides

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