Engineering the acceptor specificity of trehalose phosphorylase for the production of trehalose analogs

2012 ◽  
Vol 28 (5) ◽  
pp. 1257-1262 ◽  
Author(s):  
Jef Van der Borght ◽  
Wim Soetaert ◽  
Tom Desmet
Keyword(s):  
Acceptor Specificity ◽  
Trehalose Phosphorylase

Acceptor specificity of trehalose phosphorylase from Thermoanaerobacter brockii: Production of novel nonreducing trisaccharide, 6-O-α-D-galactopyranosyl trehalose

2006 ◽  
Vol 101 (5) ◽  
pp. 385-390 ◽  
Author(s):  
Kazuhiko Maruta ◽  
Hikaru Watanabe ◽  
Tomoyuki Nishimoto ◽  
Michio Kubota ◽  
Hiroto Chaen ◽  
...  
Keyword(s):  
Acceptor Specificity ◽  
Trehalose Phosphorylase

Acceptor specificity and tissue distribution of three human alpha-3-fucosyltransferases

1990 ◽  
Vol 191 (1) ◽  
pp. 169-176 ◽  
Author(s):  
Rosella MOLLICONE ◽  
Anne GIBAUD ◽  
Anne FRANCOIS ◽  
Murray RATCLIFFE ◽  
Rafael ORIOL
Keyword(s):  
Tissue Distribution ◽  
Acceptor Specificity

Screening of recombinant glycosyltransferases reveals the broad acceptor specificity of stevia UGT-76G1

2016 ◽  
Vol 233 ◽  
pp. 49-55 ◽  
Author(s):  
Griet Dewitte ◽  
Maarten Walmagh ◽  
Margo Diricks ◽  
Alexander Lepak ◽  
Alexander Gutmann ◽  
...  
Keyword(s):  
Acceptor Specificity

α-Retaining glucosyl transfer catalysed by trehalose phosphorylase from Schizophyllum commune: mechanistic evidence obtained from steady-state kinetic studies with substrate analogues and inhibitors

2001 ◽  
Vol 360 (3) ◽  
pp. 727-736 ◽  
Author(s):  
Bernd NIDETZKY ◽  
Christian EIS
Keyword(s):  
Steady State ◽  
Transition State ◽  
Schizophyllum Commune ◽  
Catalytic Efficiency ◽  
Kinetic Studies ◽  
Competitive Inhibitor ◽  
Chemical Mechanism ◽  
Steady State Kinetic ◽  
State Kinetic ◽  
Trehalose Phosphorylase

Fungal trehalose phosphorylase is classified as a family 4 glucosyltransferase that catalyses the reversible phosphorolysis of α,α-trehalose with net retention of anomeric configuration. Glucosyl transfer to and from phosphate takes place by the partly rate-limiting interconversion of ternary enzyme–substrate complexes formed from binary enzyme–phosphate and enzyme–α-d-glucopyranosyl phosphate adducts respectively. To advance a model of the chemical mechanism of trehalose phosphorylase, we performed a steady-state kinetic study with the purified enzyme from the basidiomycete fungus Schizophyllum commune by using alternative substrates, inhibitors and combinations thereof in pairs as specific probes of substrate-binding recognition and transition-state structure. Orthovanadate is a competitive inhibitor against phosphate and α-d-glucopyranosyl phosphate, and binds 3×104-fold tighter (Ki≈ 1μM) than phosphate. Structural alterations of d-glucose at C-2 and O-5 are tolerated by the enzyme at subsite +1. They lead to parallel effects of approximately the same magnitude (slope = 1.14; r2 = 0.98) on the reciprocal catalytic efficiency for reverse glucosyl transfer [log (Km/kcat)] and the apparent affinity of orthovanadate determined in the presence of the respective glucosyl acceptor (log Ki). An adduct of orthovanadate and the nucleophile/leaving group bound at subsite +1 is therefore the true inhibitor and displays partial transition state analogy. Isofagomine binds to subsite −1 in the enzyme–phosphate complex with a dissociation constant of 56μM and inhibits trehalose phosphorylase at least 20-fold better than 1-deoxynojirimycin. The specificity of the reversible azasugars inhibitors would be explained if a positive charge developed on C-1 rather than O-5 in the proposed glucosyl cation-like transition state of the reaction. The results are discussed in the context of α-retaining glucosyltransferase mechanisms that occur with and without a β-glucosyl enzyme intermediate.

Altering the Electron Acceptor Specificity of Flavocytochrome b2

1999 ◽  
Vol 27 (1) ◽  
pp. A45-A45
Author(s):  
Fraser Welsh ◽  
Stuart Rivers ◽  
Stephen K. Chapman ◽  
Graeme A. Reid
Keyword(s):  
Electron Acceptor ◽  
Flavocytochrome B2 ◽  
Acceptor Specificity

scholarly journals Galactosyltransferase Acceptor Specificity of the Lactose Synthetase A Protein

1970 ◽  
Vol 245 (19) ◽  
pp. 5057-5061 ◽  
Author(s):  
F.L. Schanbacher ◽  
K.E. Ebner
Keyword(s):  
Acceptor Specificity

scholarly journals Acceptor Specificity of thePasteurellaHyaluronan and Chondroitin Synthases and Production of Chimeric Glycosaminoglycans

2006 ◽  
Vol 282 (1) ◽  
pp. 337-344 ◽  
Author(s):  
Breca S. Tracy ◽  
Fikri Y. Avci ◽  
Robert J. Linhardt ◽  
Paul L. DeAngelis
Keyword(s):  
Acceptor Specificity

scholarly journals Acceptor specificity of the transglycosylation catalyzed by cyclodextrin glycosyltransferase.

1978 ◽  
Vol 42 (12) ◽  
pp. 2369-2374 ◽  
Author(s):  
Sumio KITAHATA ◽  
Shigetaka OKADA ◽  
Toshio FUKUI
Keyword(s):  
Cyclodextrin Glycosyltransferase ◽  
Acceptor Specificity

NAD+-induced phosphate acceptor specificity in submitochondrial systems

1972 ◽  
Vol 256 (2) ◽  
pp. 191-198 ◽  
Author(s):  
Ivar Vallin ◽  
Per Lundberg
Keyword(s):  
Acceptor Specificity

scholarly journals A Combination of Structural, Genetic, Phenotypic and Enzymatic Analyses Reveals the Importance of a Predicted Fucosyltransferase to Protein O-Glycosylation in the Bacteroidetes

Biomolecules ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1795
Author(s):  
Markus B. Tomek ◽  
Bettina Janesch ◽  
Matthias L. Braun ◽  
Manfred Taschner ◽  
Rudolf Figl ◽  
...  
Keyword(s):  
Structural Analysis ◽  
Mutational Analysis ◽  
Bacteroides Fragilis ◽  
Host Colonization ◽  
Acceptor Specificity ◽  
Hexose Sugars ◽  
Full Structure ◽  
Bacteroidetes Phylum ◽  
General Protein

Diverse members of the Bacteroidetes phylum have general protein O-glycosylation systems that are essential for processes such as host colonization and pathogenesis. Here, we analyzed the function of a putative fucosyltransferase (FucT) family that is widely encoded in Bacteroidetes protein O-glycosylation genetic loci. We studied the FucT orthologs of three Bacteroidetes species—Tannerella forsythia, Bacteroides fragilis, and Pedobacter heparinus. To identify the linkage created by the FucT of B. fragilis, we elucidated the full structure of its nine-sugar O-glycan and found that l-fucose is linked β1,4 to glucose. Of the two fucose residues in the T. forsythia O-glycan, the fucose linked to the reducing-end galactose was shown by mutational analysis to be l-fucose. Despite the transfer of l-fucose to distinct hexose sugars in the B. fragilis and T. forsythia O-glycans, the FucT orthologs from B. fragilis, T. forsythia, and P. heparinus each cross-complement the B. fragilis ΔBF4306 and T. forsythia ΔTanf_01305 FucT mutants. In vitro enzymatic analyses showed relaxed acceptor specificity of the three enzymes, transferring l-fucose to various pNP-α-hexoses. Further, glycan structural analysis together with fucosidase assays indicated that the T. forsythia FucT links l-fucose α1,6 to galactose. Given the biological importance of fucosylated carbohydrates, these FucTs are promising candidates for synthetic glycobiology.

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