Structural modeling of two plant UDP-dependent sugar-sugar glycosyltransferases reveals a conserved glutamic acid residue that is a hallmark for sugar acceptor recognition

Brandt, Wolfgang; Schulze, Eva; Liberman-Aloni, Raya; Bartelt, Richard; Pienkny, Silke; Carmeli-Weissberg, Mira; Frydman, Ahuva; Eyal, Yoram

doi:10.1016/j.jsb.2021.107777

Cited by 10 publications

(3 citation statements)

References 61 publications

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“…In addition to discovering additional UGTs, engineering efforts have generated UGTs with novel functions. Such advances have been assisted by structural information, which provides a basic understanding of UGT chemoselectivity [27][28][29], regioselectivity [30][31][32], sugar donor selectivity [33], and sugar acceptor selectivity [34,35]. For example, Wetterhorn et al engineered UGT Os79 from rice, which natively glycosylates a disease-causing mycotoxin, deoxynivalenol, to accommodate a bulkier analog, T-2 toxin, paving the way to protect crops against a broader scope of fungal infections [36].…”

Section: Engineering Plant Glycosyltransferasesmentioning

confidence: 99%

Harnessing Plant Sugar Metabolism for Glycoengineering

Tang,

Barnum,

Szarzanowicz

et al. 2023

Biology

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Plants possess an innate ability to generate vast amounts of sugar and produce a range of sugar-derived compounds that can be utilized for applications in industry, health, and agriculture. Nucleotide sugars lie at the unique intersection of primary and specialized metabolism, enabling the biosynthesis of numerous molecules ranging from small glycosides to complex polysaccharides. Plants are tolerant to perturbations to their balance of nucleotide sugars, allowing for the overproduction of endogenous nucleotide sugars to push flux towards a particular product without necessitating the re-engineering of upstream pathways. Pathways to produce even non-native nucleotide sugars may be introduced to synthesize entirely novel products. Heterologously expressed glycosyltransferases capable of unique sugar chemistries can further widen the synthetic repertoire of a plant, and transporters can increase the amount of nucleotide sugars available to glycosyltransferases. In this opinion piece, we examine recent successes and potential future uses of engineered nucleotide sugar biosynthetic, transport, and utilization pathways to improve the production of target compounds. Additionally, we highlight current efforts to engineer glycosyltransferases. Ultimately, the robust nature of plant sugar biochemistry renders plants a powerful chassis for the production of target glycoconjugates and glycans.

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Section: Engineering Plant Glycosyltransferasesmentioning

confidence: 99%

Harnessing Plant Sugar Metabolism for Glycoengineering

Tang,

Barnum,

Szarzanowicz

et al. 2023

Biology

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“…In tea plants (Camellia sinensis), flavonols mainly occur as O -glycosides. Glycosylation, a common modification that adds sugar moieties, results in a broad spectrum of glycosylated products . Such modification alters subcellular localization, solubility, and metabolic fates of the modified metabolites. , Furthermore, glycosylation plays a vital role in plants by regulating hormone homeostasis and conferring defense against biotic and abiotic stresses …”

Section: Introductionmentioning

confidence: 99%

Characterization of CsUGT73AC15 as a Multifunctional Glycosyltransferase Impacting Flavonol Triglycoside Biosynthesis in Tea Plants

Wang,

Sun,

et al. 2024

J. Agric. Food Chem.

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Flavonol glycosides, contributing to the health benefits and distinctive flavors of tea (Camellia sinensis), accumulate predominantly as diglycosides and triglycosides in tea leaves. However, the UDP-glycosyltransferases (UGTs) mediating flavonol multiglycosylation remain largely uncharacterized. In this study, we employed an integrated proteomic and metabolomic strategy to identify and characterize key UGTs involved in flavonol triglycoside biosynthesis. The recombinant rCsUGT75AJ1 exhibited flavonoid 4′-O-glucosyltransferase activity, while rCsUGT75L72 preferentially catalyzed 3-OH glucosylation. Notably, rCsUGT73AC15 displayed substrate promiscuity and regioselectivity, enabling glucosylation of rutin at multiple sites and kaempferol 3-O-rutinoside (K3R) at the 7-OH position. Kinetic analysis revealed rCsUGT73AC15's high affinity for rutin (K m = 9.64 μM). Across cultivars, CsUGT73AC15 expression inversely correlated with rutin levels. Moreover, transient CsUGT73AC15 silencing increased rutin and K3R accumulation while decreasing their respective triglycosides in tea plants. This study offers new mechanistic insights into the key roles of UGTs in regulating flavonol triglycosylation in tea plants.

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“…UGTs have demonstrated exceptional stereo- and regio-selectivity in glycosylation of various natural products ( Caputi et al, 2012 ; Wen et al, 2018 ; Li et al, 2020 ; Teze et al, 2021 ), showcasing their significant potential in the preparation of steviol glycoside derivatives. In our previous research ( Ping et al, 2022 ), the glycosylation of Reb A to synthesize Reb D2 through the construction of a β 1→6 glycosidic bond at the C-19 position of Reb A was facilitated by a UGT known as UGT94D1, which was originally identified from Sesamum indicum ( Noguchi et al, 2008 ; Ono et al, 2020 ; Brandt et al, 2021 ). Hence, it is plausible to achieve a highly selective and efficient synthesis of Reb M2 through the forming of a similar β 1→6 glycosidic bond using Reb D as the substrate.…”

Section: Introductionmentioning

confidence: 99%

Selective synthesis of rebaudioside M2 through structure-guided engineering of glycosyltransferase UGT94D1

Yang,

Deng

et al. 2024

Front. Bioeng. Biotechnol.

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Rebaudioside M2 (Reb M2), a novel steviol glycoside derivative, has limited industrial applications due to its low synthetic yield and selectivity. Herein, we identify UGT94D1 as a selective glycosyltransferase for rebaudioside D (Reb D), leading to the production of a mono β-1,6-glycosylated derivative, Reb M2. A variant UGT94D1-F119I/D188P was developed through protein engineering. This mutant exhibited a 6.33-fold improvement in catalytic efficiency, and produced Reb M2 with 92% yield. Moreover, molecular dynamics simulations demonstrated that UGT94D1-F119I/D188P exhibited a shorter distance between the nucleophilic oxygen (OH6) of the substrate Reb D and uridine diphosphate glucose, along with an increased Ophosphate-C1-Oacceptor angle, thus improving the catalytic activity of the enzyme. Therefore, this study provides an efficient method for the selective synthesis of Reb M2 and paves the way for its applications in various fields.

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Structural modeling of two plant UDP-dependent sugar-sugar glycosyltransferases reveals a conserved glutamic acid residue that is a hallmark for sugar acceptor recognition

Cited by 10 publications

References 61 publications

Harnessing Plant Sugar Metabolism for Glycoengineering

Harnessing Plant Sugar Metabolism for Glycoengineering

Characterization of CsUGT73AC15 as a Multifunctional Glycosyltransferase Impacting Flavonol Triglycoside Biosynthesis in Tea Plants

Selective synthesis of rebaudioside M2 through structure-guided engineering of glycosyltransferase UGT94D1

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