Cloning and characterization of a glucosyltransferase that reacts on 7-hydroxyl group of flavonol and 3-hydroxyl group of coumarin from tobacco cells

Taguchi, Goro; Ubukata, Takahisa; Hayashida, Nobuaki; Yamamoto, Haruhiko; Okazaki, Mitsuo

doi:10.1016/j.abb.2003.09.027

Cited by 37 publications

(33 citation statements)

References 29 publications

(48 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is noteworthy that His-15* and Asp-125* are replaced by other amino acid residues in some glycosyltransferases; for example, His-15* is replaced by cysteine in Arabidopsis thaliana UGT76C1 (DDBJ/GenBank TM /EBI accession number BAB10792) and UGT76C2 (accession number AAN28835) (see Ref. 20), and Asp-125* is replaced by arginine in Crocus sativus UGTCs2 (accession number AAP94878), serine in Nicotiana tabacum glucosyltransferase-2 (accession number BAB88935) (39) and Petunia ϫ hybrida UDP-glucose:anthocyanin 5-O-glucosyltransferase (accession number BAA89009) (Fig. 4), and asparagine in Brassica napus UDP-glucose:sinapate glucosyltransferase (accession number AAF98390) (40).…”

Section: Exploring Amino Acid Residues That Are Important For Gmif7gtmentioning

confidence: 99%

A UDP-Glucose:Isoflavone 7-O-Glucosyltransferase from the Roots of Soybean (Glycine max) Seedlings

Niimi

Saito

Homma

et al. 2007

Journal of Biological Chemistry

117

View full text Add to dashboard Cite

Isoflavones, a class of flavonoids, play very important roles in plant-microbe interactions in certain legumes such as soybeans (Glycine max L. Merr.). G. max UDP-glucose:isoflavone 7-Oglucosyltransferase (GmIF7GT) is a key enzyme in the synthesis of isoflavone conjugates, which accumulate in large amounts in vacuoles and serve as an isoflavonoid pool that allows for interaction with microorganisms. In this study, the 14,000-fold purification of GmIF7GT from the roots of G. max seedlings was accomplished. The purified enzyme is a monomeric protein of 46 kDa, catalyzing regiospecific glucosyl transfer from UDPglucose to isoflavones to produce isoflavone 7-O-␤-D-glucosides (k cat ‫؍‬ 0.74 s ؊1 , K m for genistein ‫؍‬ 3.6 M, and K m for UDPglucose ‫؍‬ 190 M). The GmIF7GT cDNA was isolated based on the amino acid sequence of the purified enzyme. Phylogenetic analysis showed that GmIF7GT is a novel member of glycosyltransferase family 1 and is distantly related to Glycyrrhiza echinata UDP-glucose:isoflavonoid 7-O-glucosyltransferase. The purified enzyme was unexpectedly devoid of the N-terminal 49-residue segment and thus lacks the histidine residue corresponding to the proposed catalytic residue of glycosyltransferases from Medicago truncatula (UGT71G1) and Vitis vinifera (VvGT1). The results of kinetic studies of site-directed mutants of GmIF7GT showed that both His-15 and Asp-125, which correspond to the catalytic residues of UGT71G1 and VvGT1, are not important for GmIF7GT activity. The results also suggest that an acidic residue at position 392 is very important for primary catalysis of GmIF7GT. These results led to the proposal that GmIF7GT utilizes a strategy of catalysis that is distinct from those proposed for UGT71G1 and VvGT1.

show abstract

Section: Exploring Amino Acid Residues That Are Important For Gmif7gtmentioning

confidence: 99%

A UDP-Glucose:Isoflavone 7-O-Glucosyltransferase from the Roots of Soybean (Glycine max) Seedlings

Niimi

Saito

Homma

et al. 2007

Journal of Biological Chemistry

117

View full text Add to dashboard Cite

show abstract

“…Glycosyltransferases UGT73C8 from Medicago truncatula glycosylate non-flavonoid substrates (Modolo et al 2007). NtGT2 from tobacco glycosylates not only the flavonoid at 7-OH, but also coumarin at 3-OH (Taguchi et al 2003). Medicago UGT71G1 and UGT73C8 expressed in E. coli glycosylate flavones and isoflavones (He et al 2008), while UGT73F1 from Glycyrrhiza echinata shows isoflavonespecific activity (Nagashima et al 2004).…”

Section: Introductionmentioning

confidence: 97%

“…Flavonoid glycosides are the major constituents found in the extracts of S. baicalensis Georgi and S. barbata D. Don (Parajuli et al 2009). Glycosylation, which is the last step in the biosynthesis of many plant secondary metabolites, affects the properties of solubility, bioavailability and detoxification of aglycones (Vogt and Jones 2000;Taguchi et al 2003). Plant UDP glycosyltransferases (UGTs,EC 2.4.1.x) belong to the family 1 glycosyltransferase (GTs, EC 2.4.x.y) based on the presence of a highly conserved plant secondary product glucosyltransferase signature (PSPG box) that consists of 44 amino acids in the C-terminal region as the nucleotide-sugar binding site (Hughes and Hughes 1994).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the Scutellaria barbata glycosyltransferase gene and its promoter

Chiou

Fang

et al. 2010

Planta

View full text Add to dashboard Cite

The conversion of flavonoid aglycones to their glycosides by plant glycosyltransferases may affect a wide range of outcomes, including stability, solubility and bioavailability. Scutellaria barbata, rich in flavonoid glycosides, is widely used as a traditional Chinese herbal medicine. In this study, a flavonoid glycosyltransferase cDNA (SbUGT) and its promoter from S. barbata were cloned and characterized as a flavonoid glycosyltransferase using whole-cell biotransformation. Fragments of different lengths of the 5'-flanking region of the SbUGT gene were fused to the beta-glucuronidase (GUS) gene and analyzed with transgenic Arabidopsis plants using histochemical and fluorometric assays. GUS activity in transgenic plants carrying the SbP-850U construct (-850 to +86 relative to the transcription start site) displayed the highest level and was enhanced by salt and methyl jasmonate, similar to the expression patterns of the endogenous SbUGT. GUS activity disappeared when the promoter was deleted to -98, and deletion analyses indicated the existence of positive and negative regulatory element(s). Unexpectedly, plants carrying the construct SbP-102U (-102 to +86) exhibited strong GUS activity exclusively in the roots. Our experiments revealed that the specific expression is mediated by different promoter regions and the unique region driving root-preferred expression can be used as a root-specific promoter.

show abstract

“…Additionally, there are several reports that plant hormone treatments can alter GTase expression patterns (Taguchi et al 2001;Taguchi et al 2003a;Taguchi et al 2003b). Thus, we hypothesize that the regulation of PNgt1 expression may be correlated to the plant hormone treatment.…”

Section: Expression Of Pngt1 In Pnhr Treated With Plant Hormonesmentioning

confidence: 99%

“…However, the GTase related to detoxification, defense and phytoalexin production has only been investigated in a few plants. Interestingly, the expression levels of GTases related to detoxification are altered by MeJA and salicylic acid (SA) (Taguchi et al 2001;Taguchi et al 2003a;Taguchi et al 2003b). These plant hormones are closely related to the activation or inactivation of plant defense systems (Pieterse et al 2009), and the results suggest that GTases will play key roles in the plant defense system.…”

mentioning

confidence: 99%

Molecular cloning and characterization of coumarin glucosyltransferase in hairy roots of Pharbitis nil (Ipomoea nil)

Kanoh

Kawauchi

Kuroyanagi

et al. 2014

Plant Biotechnology

View full text Add to dashboard Cite

The hairy roots induced from Pharbitis nil (Ipomoea nil) by infection of the plant pathogen Agrobacterium rhizogenes produced and released the phytoalexin of umbelliferone into the culture medium by treatment with copper sulfate. However, this umbelliferone was immediately recovered in the hairy roots as its glucoside, skimmin, after a few hours. To analyze the details of these phenomena, we searched for glucosyltransferase (GTase) genes related to skimmin production and isolated two full-length cDNAs, named PNgt1 and PNgt2. They contain the plant secondary product glucosyltransferase (PSPG) box that is a common signature of plant GTases. Phylogenetic analyses of PNgt1 and PNgt2 indicated that they encode a broad substrate-specific GTase and an unclassified GTase, respectively. An enzyme assay of the recombinant PNGT1 protein showed a high glucosylation rate against coumarin and benzaldehyde derivatives, especially umbelliferone. The expression level of PNgt1 after treatment with umbelliferone showed a correlation with skimmin production. Moreover, we found the expression levels of PNgt1 were altered by the plant hormones salicylic acid and methyl jasmonate. These results suggest the importance of plant GTases in phytoalexin production and plant defense mechanisms.

show abstract

Cloning and characterization of a glucosyltransferase that reacts on 7-hydroxyl group of flavonol and 3-hydroxyl group of coumarin from tobacco cells

Cited by 37 publications

References 29 publications

A UDP-Glucose:Isoflavone 7-O-Glucosyltransferase from the Roots of Soybean (Glycine max) Seedlings

A UDP-Glucose:Isoflavone 7-O-Glucosyltransferase from the Roots of Soybean (Glycine max) Seedlings

Characterization of the Scutellaria barbata glycosyltransferase gene and its promoter

Molecular cloning and characterization of coumarin glucosyltransferase in hairy roots of Pharbitis nil (Ipomoea nil)

Contact Info

Product

Resources

About