A Glycosynthase Catalyst for the Synthesis of Flavonoid Glycosides

Yang, Min; Davies, G.J.; Davis, Benjamin G.

doi:10.1002/anie.200604177

Cited by 76 publications

(56 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A recent study, using a high-throughput screen to detect novel donor and acceptor activities, revealed the surprising ability of a Humicola insolens Cel7B glycosynthase mutant (35) to glycosylate flavonoid scaffolds (36). Interestingly, the glycosynthase showed a Ͼ95% regiospecificity toward the 4Ј-OH of quercetin using the activated donor lactosyl fluoride.…”

Section: Discussionmentioning

confidence: 99%

A Kinetic Analysis of Regiospecific Glucosylation by Two Glycosyltransferases of Arabidopsis thaliana

Cartwright

Lim

Kleanthous

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

Plant Family 1 glycosyltransferases (GTs) recognize a wide range of natural and non-natural scaffolds and have considerable potential as biocatalysts for the synthesis of small molecule glycosides. Regiospecificity of glycosylation is an important property, given that many acceptors have multiple potential glycosylation sites. This study has used a domain-swapping approach to explore the determinants of regiospecific glycosylation of two GTs of Arabidopsis thaliana, UGT74F1 and UGT74F2. The flavonoid quercetin was used as a model acceptor, providing five potential sites for O-glycosylation by the two GTs. As is commonly found for many plant GTs, both of these enzymes produce distinct multiple glycosides of quercetin. A high performance liquid chromatography method has been established to perform detailed steady-state kinetic analyses of these concurrent reactions. These data show the influence of each parameter in determining a GT product formation profile toward quercetin. Interestingly, construction and kinetic analyses of a series of UGT74F1/F2 chimeras have revealed that mutating a single amino acid distal to the active site, Asn-142, can lead to the development of a new GT with a more constrained regiospecificity. This ability to form the 4-O-glucoside of quercetin is transferable to other flavonoid scaffolds and provides a basis for preparative scale production of flavonoid 4-O-glucosides through the use of wholecell biocatalysis.The glycosylation of small hydrophobic molecules is known to alter their chemical and biological characteristics. The alterations include changes in water solubility, stability, pharmacokinetic properties, and bioactivity (1, 2). Enzymes that catalyze these reactions are Family 1 glycosyltransferases (GTs), 3 with representatives found in a wide range of prokaryotic and eukaryotic organisms.Extensive study of GT activities has demonstrated that the enzymes display an exquisite regio-(3), enantio-(4), and chemospecificity (5) toward the acceptor molecule. However, as yet the determinants of this specificity are poorly understood. This lack of understanding presents challenges to the interpretation of substrate activity data (6, 7), GT rational redesign (8, 9), and activity prediction (10, 11). Future progress in predicting GT sequence-structure-activity relationships will depend on a greater number of studies that characterize the key determinants of activity and specificity.Quercetin is the most abundant flavonoid in the human diet (12) and has frequently been used as a model substrate for GT activity (13-15). Quercetin aglycone is found only at low concentrations in the primary dietary source, plants, where glycosylated forms predominate (16). In addition, in mammals, glycosides are the major byproducts of quercetin phase II metabolism (17). Many studies have shown that glycosylation of quercetin significantly affects bioavailability and efficacy with respect to anti-oxidant, anti-proliferative, and anti-cancer properties (18). There are also some data to indicate that the positi...

show abstract

Section: Discussionmentioning

confidence: 99%

A Kinetic Analysis of Regiospecific Glucosylation by Two Glycosyltransferases of Arabidopsis thaliana

Cartwright

Lim

Kleanthous

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…It is worthwhile to note in this context that the E197S mutant of the Humicola insolens Cel7B glycosidase catalyzed the synthesis of flavonoid glycosides, transferring the disaccharide donor lactosyl fluoride to several flavonoids. [20] The enzymatic transglycosylation of compound 16, which contains three phenolic groups, yielded a mixture of two regioisomeric glycosides in the ratio of 2:1 16a:16b (C-3 and C-4' glycosides). Preparation of rutinosides using transglycosylations of aliphatic alcohols have been described in the past with a crude extract of Fagopyrum tataricum seeds; [1] here, methyl-, ethyl-, propyl-, isopropyl-and benzyl-b-rutinosides were obtained in relatively high yields (84-94%).…”

Section: Enzymatic Synthesis Of B-rutinosidesmentioning

confidence: 99%

α‐L‐Rhamnosyl‐β‐D‐glucosidase (Rutinosidase) from Aspergillus niger: Characterization and Synthetic Potential of a Novel Diglycosidase

et al. 2014

View full text Add to dashboard Cite

Abstract:We report the first heterologous production of a fungal rutinosidase (6-O-a-l-rhamnopyranosyl-b-d-glucopyranosidase) in Pichia pastoris. The recombinant rutinosidase was purified from the culture medium to apparent homogeneity and biochemically characterized. The enzyme reacts with rutin and cleaves the glycosidic linkage between the disaccharide rutinose and the aglycone. Furthermore, it exhibits high transglycosylation activity, transferring rutinose from rutin as a glycosyl donor onto various alcohols and phenols. The utility of the recombinant rutinosidase was demonstrated by its use for the synthesis of a broad spectrum of rutinosides of primary (saturated and unsaturated), secondary, acyclic and phenolic alcohols as well as for the preparation of free rutinose. Moreover, the a-l-rhamnosidase-catalyzed synthesis of a chromogenic substrate for a rutinosidase assay -para-nitrophenyl b-rutinoside -is described.

show abstract

“…Likewise, glycosylated small-molecule natural products are accessible by the glycosynthase technology. The broad acceptor specificity of the glycosynthase mutant E197S from cellulase Cel7B has shown that it can directly glycosylate flavonoids with high efficiency [87].…”

Section: Glycosynthases From Retaining Gh With Enzyme Nucleophile Andmentioning

confidence: 99%

When Enzymes Do It Better: Enzymatic Glycosylation Methods

Planas¹,

Faijes²,

Codera³

2015

Carbohydrate Chemistry: State of the Art and Challenges for Drug Development

View full text Add to dashboard Cite

Glycoside Bond Formation by CAZymesCarbohydrates, in the forms of oligosaccharides, polysaccharides, and glycoconjugates (glycoproteins, glycolipids, glycosylated metabolites, etc) are key biomolecules in all living organisms. They play roles in a variety of biological functions as structural components of cell walls, reserves of stored energy, and mediators of cell-cell interactions, being central components in many important biological recognition processes, such as cell adhesion and signaling, cancer progression, host-pathogen interactions, and immune responses [1][2][3][4][5].Synthetic tools to access natural and non-natural glycosides and glycoconjugates have become a central issue in glycochemistry and glycobiology. Both for industrial applications in diverse fields, such as medicine, pharmacology, food, cosmetics, etc., as well as for biological research in functional glycomic studies, there is the need for efficient synthetic approaches for structurally defined oligosaccharides and glycoconjugates. In contrast to conventional chemical synthesis that requires complex and tedious protection/deprotection manipulations and activation strategies to achieve regio-and Carbohydrate Chemistry: State of the Art and Challenges for Drug Development Downloaded from www.worldscientific.com by UNIVERSITY OF PITTSBURGH on 10/02/15. For personal use only.

show abstract

A Glycosynthase Catalyst for the Synthesis of Flavonoid Glycosides

Cited by 76 publications

References 34 publications

A Kinetic Analysis of Regiospecific Glucosylation by Two Glycosyltransferases of Arabidopsis thaliana

A Kinetic Analysis of Regiospecific Glucosylation by Two Glycosyltransferases of Arabidopsis thaliana

α‐L‐Rhamnosyl‐β‐D‐glucosidase (Rutinosidase) from Aspergillus niger: Characterization and Synthetic Potential of a Novel Diglycosidase

When Enzymes Do It Better: Enzymatic Glycosylation Methods

Contact Info

Product

Resources

About