Molecular and Structural Characterization of a Promiscuous <i>C</i>‐Glycosyltransferase from <i>Trollius chinensis</i>

He, Junbin; Zhao, Peng; Hu, Zongmin; Liu, Shuang; Kuang, Yi; Zhang, Meng; Li, Bin; Yun, Cai‐Hong; Qiao, Xue; Ye, Min

doi:10.1002/anie.201905505

Cited by 111 publications

(121 citation statements)

References 54 publications

Supporting

Mentioning

115

Contrasting

Order By: Relevance

“…CGTs belong to the family of the UDP-dependent glycosyltransferases (UGTs) that utilize activated UDP-sugars as donor. Despite that direct C-glycosylation on the final flavonoid skeleton have been documented in Pueraria lobate 25 , Gentiana triflora 26 and Trollius chinensis 27 , the other existing CGTs exclusively come from UGT708 subfamily and glycosylate 2hydroxylated intermediates with UDP-glucose (UDP-Glc). Considering the chemical diversity of known CGFs bearing various sugars and different glycosylation patterns other than glucosyl substitution, there is still a huge gap between known CGTs and diverse structures of CGFs.…”

mentioning

confidence: 99%

Pathway-specific enzymes from bamboo and crop leaves biosynthesize anti-nociceptive C-glycosylated flavones

et al. 2020

View full text Add to dashboard Cite

C-glycosylated flavones (CGFs) are promising candidates as anti-nociceptive compounds. The leaves of bamboo and related crops in the grass family are a largely unexploited bioresource with a wide array of CGFs. We report here pathway-specific enzymes including Cglycosyltransferases (CGTs) and P450 hydroxylases from cereal crops and bamboo species accumulating abundant CGFs. Mining of CGTs and engineering of P450s that decorate the flavonoid skeleton allowed the production of desired CGFs (with yield of 20-40 mg/L) in an Escherichia coli cell factory. We further explored the antinociceptive activity of major CGFs in mice models and identified isoorientin as the most potent, with both neuroanalgesic and antiinflammatory effects superior to clinical drugs such as rotundine and aspirin. Our discovery of the pain-alleviating flavonoids elicited from bamboo and crop leaves establishes this previously underutilized source, and sheds light on the pathway and pharmacological mechanisms of the compounds.

show abstract

mentioning

confidence: 99%

Pathway-specific enzymes from bamboo and crop leaves biosynthesize anti-nociceptive C-glycosylated flavones

et al. 2020

View full text Add to dashboard Cite

show abstract

“…TcCGT1 was identified to catalyze four types of glycosylation, the C-or O-position of phenols and flavonoids, the N-position of 3,4-dichloraaniline, and S-position of 3,4chlorothiophenol (He et al, 2019). In contrast, GbNGT1 specifically catalyzes the Nglucosylation toward IAA or IAA-AA.…”

Section: Specificity Of N-glucosylationmentioning

confidence: 99%

“…So far, very few key residues in N terminal were identified even if some crystal structures of NGTs were revealed. H24 and E396 of TcCGT1 were demonstrated to play key role in the stabilization and location of small molecule substrates; in addition, the mutants I94E and G284K of TcCGT1 could convert enzymatic activity from C-to O-position (He et al, 2019).…”

Section: Specificity Of N-glucosylationmentioning

confidence: 99%

“…In the meanwhile, it could also form O-and N-glycosides (Chen et al, 2015). In medicinal plant, TcCGT1 (Trollius chinensis) was confirmed to catalyze C-, O-, N-, and S-glycosylation reactions (He et al, 2019). The substrates specificity of glycosyltransfeases mainly depend on their structures, rather than their preliminary sequences (Ostrowski et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

N-glucosyltransferase GbNGT1 from Ginkgo complement auxin metabolic pathway

Yin

Zhang

Wang

et al. 2020

Preprint

View full text Add to dashboard Cite

As a group of the most important phytohormone, auxin homeostasis is regulated in a complex manner. Generally, auxin conjugations especially IAA glucosides are dominant on high auxin level conditions. Former terminal glucosylation researches mainly focus on O-position, while IAA-N-glucoside or IAA-Asp-N-glucoside has been neglected since their found in 2001. In our study, IAA-Asp-N-glucoside was firstly found specifically abundant (as high as 4.13 mg/g) in ginkgo seeds of 58 cultivars from Ginkgo Resource Nursery built in 1990. Furthermore, a novel N-glucosyltransferase GbNGT1, which could catalyze IAA-Asp and IAA to form their corresponding N-glucoside, was identified through differential transcriptome analysis and in vitro enzymatic test. The enzyme was demonstrated to possess specific catalyze capacity toward the N-position of IAA-amino acid or IAA among 52 substrates, and was typical of acid tolerance, metal ion independence and high temperature sensitivity. Docking and site-directed mutagenesis of this enzyme confirmed that E15G mutant could almost abolish enzyme catalytic activity towards IAA-Asp and IAA in vitro and in vivo. The IAA modification of GbNGT1 and GbGH3.5 was verified by transient expression assay in Nicotiana benthamiana. In conclusion, our results complement the terminal metabolic pathway of auxin, and the specific catalytic function of GbNGT1 towards IAA-amino acid provide a new way to biosynthesis indole-amide compounds.HighlightThe N-glucosylation of IAA or IAA-amino acids in auxin metabolism had been neglected over decades, our work for GbNGT1 redeems the missing chain of auxin metabolic pathway.

show abstract

“…Subsequently, many UGT structures have been determined with or without different sugar acceptors, such as PtUGT1 from Polygonum tinctorium with indoxyl sulfate (Hsu et al, 2018), UGT78K6 from Clitoria ternatea with anthocyanins (Hiromoto et al, 2015), UGT74F2 from Arabidopsis thaliana with sialic acid (George Thompson et al, 2017) and Os79 from Oryza sativa with trichothecene (Wetterhorn et al, 2016). Recently, the crystal structures of UGT76G1 from Stevia rebaudiana, which glycosylates the larger acceptor steviol (Yang et al, 2019), UGT89C1 from A. thaliana, which utilizes UDP-rhamnose as a sugar donor (Zong et al, 2019), and the C-glycosyltransferase TcCGT1 from Trollius chinensis (He et al, 2019) have been determined, which widened our knowledge of the catalytic mechanism in UGTs. However, the diversity in the polyphenols, the ability of UGTs to recognize specific compounds, the regioselectivity of products and the sugar-donor specificity of UGTs necessitates the study and determination of the crystal structures of more UGTs in order to understand the substrate recognition and the regioselectivity of the product.…”

Section: Introductionmentioning

confidence: 99%

Crown-ether-mediated crystal structures of the glycosyltransferasePaGT3 fromPhytolacca americana

Maharjan

Fukuda

Nakayama

et al. 2020

Acta Cryst Sect D Struct Biol

View full text Add to dashboard Cite

Uridine diphosphate glycosyltransferases (UGTs) are ubiquitous enzymes that are involved in the glycosylation of small molecules. As glycosylation improves the water solubility and stability of hydrophobic compounds, interest in the use of UGTs for the synthesis of glycosides of poorly soluble compounds is increasing. While sugar-donor recognition in UGTs is conserved with the presence of a plant secondary product glycosyltransferase (PSPG) motif, the basis of the recognition of the sugar acceptor and the regioselectivity of the products is poorly understood owing to low sequence identity around the acceptor-binding region. PaGT3, a glycosyltransferase from the plant Phytolacca americana, can glycosylate a range of acceptors. To illustrate the structurefunction relationship of PaGT3, its crystal structure was determined. The sugardonor and sugar-acceptor binding pockets in PaGT3 were recognized by comparison of its structure with those of other UGTs. The key feature of PaGT3 was the presence of longer loop regions around the hydrophobic acceptorbinding pocket, which resulted in a flexible and wider acceptor binding pocket. In this study, PaGT3 crystals were grown by co-crystallization with 18-crown-6 ether or 15-crown-5 ether. The crown-ether molecule in the asymmetric unit was observed to form a complex with a metal ion, which was coordinated on two sides by the main-chain O atoms of Glu238 from two molecules of the protein.The crown ether-metal complex resembles a molecular glue that sticks two molecules of PaGT3 together to enhance crystal growth. Thus, this result provides an insight into the substrate-recognition strategy in PaGT3 for the study of glycosyltransferases. Additionally, it is shown that crown ether-metal ion complexes can be used as a molecular glue for the crystallization of proteins.

show abstract

Molecular and Structural Characterization of a Promiscuous C‐Glycosyltransferase from Trollius chinensis

Cited by 111 publications

References 54 publications

Pathway-specific enzymes from bamboo and crop leaves biosynthesize anti-nociceptive C-glycosylated flavones

Pathway-specific enzymes from bamboo and crop leaves biosynthesize anti-nociceptive C-glycosylated flavones

N-glucosyltransferase GbNGT1 from Ginkgo complement auxin metabolic pathway

Crown-ether-mediated crystal structures of the glycosyltransferasePaGT3 fromPhytolacca americana

Contact Info

Product

Resources

About