Probing the transferase activity of glycosidases by means of in situ NMR spectroscopy

Spangenberg, Petra; Chiffoleau-Giraud, Violaine; André, Corinne; Dion, Michel; Rabiller, Claude

doi:10.1016/s0957-4166(99)00297-9

Cited by 19 publications

(10 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The α-1-3 pure disaccharide was further transformed attaching a PEG (poly-ethyleneglycol) molecule in order to increase molecular size for pharmacokinetic improvements [39]. The dimerization of the substrate pnitrophenyl α-D-galactopyranoside with coffe beans α-galactosidase was also used to probe the reaction for testing a new technique for the quick analysis of the enzymatic regioselectivity using NMR spectroscopy [40].…”

Section: Glycosyl Hydrolases α-Galactosidasesmentioning

confidence: 99%

See 1 more Smart Citation

Glycosyl Hydrolases and Glycosyltransferases in the Synthesis of Oligosaccharides

Trincone¹,

Giordano

2006

COC

View full text Add to dashboard Cite

Glycobiology and related disciplines have received an enormous interest in recent years as they shed new light on the functional roles of carbohydrates in biological events leading to the understanding of mechanisms of important pathologies and to the development of new therapeutics. Although carbohydrate can be isolated from natural sources, the synthetic strategy plays its own role allowing access to larger quantities of structurally defined material and entry to analogs of naturally occurring structures. Nowadays, the bottleneck in this field is represented by some limitations of the potential of carbohydrate containing molecules because their complex structures make classical chemical synthesis very difficult.The synthesis of oligosaccharides have to face with three main snags: (i) reactivity of the leaving group on the monosaccharide acting as donor; (ii) regioselectivity towards a single hydroxyl group on the acceptor molecule; (iii) stereoselectivity in forming pure anomers of the new glycosidic product. The protecting-group manipulations that are needed for the stereocontrol of the products are demanding procedures thus yields and selectivity are lowered hindering the efficient production of oligosaccharides needed for biological testing.A particular benefit of the renewed interest for sugar chemistry has been the attention of scientific community to the biological methodologies in the production of oligosaccharides, an area which emerged in recent decades.In the carbohydrate field different enzymes were used for diverse purposes but enzymatic strategies for high-yield and stereospecific construction of glycosidic bonds are based on the action of two types of enzymes: glycosyl hydrolases (endo-and exo-glycosidases) and glycosyltransferases. Glycosyl hydrolases in the cells are responsible for the cleavage of glycosidic linkages; the exo-glycosidase are involved for glycan processing during in vivo glycoprotein synthesis. The glycosyltransferases are instead responsible in vivo for the synthesis of most cell-surface glycoconjugates.This review deals with the application of different types of glycosyl hydrolases. Elegant examples concerning the use of genetically modified representatives of glycosyl hydrolases (glycosynthases and thioglycoligases) will be also reported; some recent advances on the use of glycosyltransferases are also included.In compiling this review we were aware of the huge amount of excellent material published in the recent years on this topic, thus we will limit the covering of literature to the last decade. Attention will be devoted to the regioselectivity towards pyranosidic templates and to the yield of reactions. The molecular diversity obtained by the use of enzymes from different biological sources and the biological importance of the compounds synthesized will be focused as well.This review will put in evidence that glycosyl hydrolases and glycosyltransferases for the synthesis of the glycosidic linkages will play a relevant role in the next future as on the bench bio-catalysts ...

show abstract

Section: Glycosyl Hydrolases α-Galactosidasesmentioning

confidence: 99%

“…Different thermophilic α-galactosidases were also screened by in situ NMR spectroscopy technique [40]. They are isolated from Bacillus stearothermophilus (Aga A, Aga B and Aga 285), from Thermus brockanius, from Streptococcus mutans and from E. coli, strain D1021.…”

Section: Glycosyl Hydrolases α-Galactosidasesmentioning

confidence: 99%

Glycosyl Hydrolases and Glycosyltransferases in the Synthesis of Oligosaccharides

Trincone¹,

Giordano

2006

COC

View full text Add to dashboard Cite

show abstract

“…Moreover, when working with an exo-glycosidase, two types of glycosides were usually synthesized: those resulting from the reaction between donor and acceptor and those produced from the self-condensation of the donor. [1][2][3] An impressive advancement of the method was achieved after the elucidation of the mechanism of retaining β-glucosidases by Withers et al [4][5][6] A pair of carboxylic acids acting as catalytic residues were shown to be involved in the process. One conditions, under which the rate of spontaneous hydrolysis of the donor was considerably lower than that of enzymatic transglycosidation, provided galactosyl and glucosyl β-(1Ǟ3)-glycosides in yields of up to 90 %.…”

Section: Introductionmentioning

confidence: 99%

Thermus thermophilus Glycosynthases for the Efficient Synthesis of Galactosyl and Glucosyl β‐(1→3)‐Glycosides

et al. 2005

View full text Add to dashboard Cite

Inverting mutant glycosynthases were designed according to the Withers strategy, starting from wild-type Thermus thermophilus retaining Tt-β-Gly glycosidase. Directed mutagenesis of catalytic nucleophile glutamate 338 by alanine, serine, and glycine afforded the E338A, E338S, and E338G mutant enzymes, respectively. As was to be expected, the mutants were unable to catalyze the hydrolysis of the transglycosidation products. In agreement with previous results, the E338S and E338G catalysts were much more efficient than E338A. Moreover, our results showed that these enzymes were inactive in the hydrolysis of the α-D-glycopyranosyl fluorides used as donors, and so suitable experimental

show abstract

“…1 H nuclear magnetic resonance (NMR) spectroscopy is an established technique for the study of enzyme kinetics that has been used to characterise different enzyme systems including (but not limited to) carbohydrate-processing enzymes, 23 , 24 enzymes related to antibiotic resistance 25 and oxygenases. 26 , 27 1 H NMR spectroscopy enables the direct monitoring of reaction kinetics in real time and accurate, quantitative information can be obtained by following changes in the peak area of the resonances associated with the substrate and/or reaction product(s).…”

Section: Introductionmentioning

confidence: 99%

Development of NMR and thermal shift assays for the evaluation ofMycobacterium tuberculosisisocitrate lyase inhibitors

et al. 2017

View full text Add to dashboard Cite

show abstract

Probing the transferase activity of glycosidases by means of in situ NMR spectroscopy

Cited by 19 publications

References 17 publications

Glycosyl Hydrolases and Glycosyltransferases in the Synthesis of Oligosaccharides

Glycosyl Hydrolases and Glycosyltransferases in the Synthesis of Oligosaccharides

Thermus thermophilus Glycosynthases for the Efficient Synthesis of Galactosyl and Glucosyl β‐(1→3)‐Glycosides

Development of NMR and thermal shift assays for the evaluation ofMycobacterium tuberculosisisocitrate lyase inhibitors

Contact Info

Product

Resources

About