173468-29-6Relevant articles and documents
A new archaeal β-glycosidase from Sulfolobus solfataricus: Seeding a novel retaining β-glycan-specific glycoside hydrolase family along with the human non-lysosomal glucosylceramidase GBA
Cobucci-Ponzano, Beatrice,Aurilia, Vincenzo,Riccio, Gennaro,Henrissat, Bernard,Coutinho, Pedro M.,Strazzulli, Andrea,Padula, Anna,Corsaro, Maria Michela,Pieretti, Giuseppina,Pocsfalvi, Gabriella,Fiume, Immacolata,Cannio, Raffaele,Rossi, Mose,Moracci, Marco
, p. 20691 - 20703 (2010)
Carbohydrate active enzymes (CAZymes) are a large class of enzymes, which build and breakdown the complex carbohydrates of the cell.Onthe basis of their amino acid sequences they are classified in families and clans that show conserved catalytic mechanism, structure, and active site residues, but may vary in substrate specificity. We report here the identification and the detailed molecular characterization of a novel glycoside hydrolase encoded from the gene sso1353 of the hyperthermophilic archaeon Sulfolobus solfataricus. This enzyme hydrolyzes aryl β-gluco- and β-xylosides and the observation of transxylosylation reactions products demonstrates that SSO1353 operates via a retaining reaction mechanism. The catalytic nucleophile (Glu-335) was identified through trapping of the 2-deoxy-2-fluoroglucosyl enzyme intermediate and subsequent peptide mapping, while the general acid/base was identified as Asp-462 through detailed mechanistic analysis of a mutant at that position, including azide rescue experiments. SSO1353 has detectable homologs of unknown specificity among Archaea, Bacteria, and Eukarya and shows distant similarity to the non-lysosomal bile acid β-glucosidase GBA2 also known as glucocerebrosidase. On the basis of our findings we propose that SSO1353 and its homologs are classified in a new CAZy family, named GH116, which so far includes β-glucosidases (EC 3.2.1.21), β-xylosidases (EC 3.2.1.37), and glucocerebrosidases (EC 3.2.1.45) as known enzyme activities.
Novel substrates for the automated and manual assay of endo-1,4-β-xylanase
Mangan, David,Cornaggia, Claudio,Liadova, Agnija,McCormack, Niall,Ivory, Ruth,McKie, Vincent A.,Ormerod, Aaron,McCleary, Barry V.
, p. 14 - 22 (2017)
endo-1,4-β-Xylanase (EC 3.2.1.8) is employed across a broad range of industries including animal feed, brewing, baking, biofuels, detergents and pulp (paper). Despite its importance, a rapid, reliable, reproducible, automatable assay for this enzyme that is based on the use of a chemically defined substrate has not been described to date. Reported herein is a new enzyme coupled assay procedure, termed the XylX6 assay, that employs a novel substrate, namely 4,6-O-(3-ketobutylidene)-4-nitrophenyl-β-45-O-glucosyl-xylopentaoside. The development of the substrate and associated assay is discussed here and the relationship between the activity values obtained with the XylX6 assay versus traditional reducing sugar assays and its specificity and reproducibility were thoroughly investigated.
Glycosynthase-Mediated Assembly of Xylanase Substrates and Inhibitors
Goddard-Borger, Ethan D.,Fiege, Brigitte,Kwan, Emily M.,Withers, Stephen G.
experimental part, p. 1703 - 1711 (2012/06/29)
An exo-β-xylosidase mutant with glycosynthase activity was created to aid in the synthesis of xylanase substrates and inhibitors. Simple monosaccharides were easily elaborated into di-, tri- and tetrasaccharides by using this enzyme. Some products proved
