title: Distinct Substrate Specificity and Catalytic Activity of the Pseudoglycosyltransferase VldE.
abstract: The pseudoglycosyltransferase (PsGT) VldE is a glycosyltransferase-like protein that is similar to trehalose 6-phosphate synthase (OtsA). However, in contrast to OtsA, which catalyzes condensation between UDP-glucose and glucose 6-phosphate, VldE couples two pseudosugars to give a product with an α,α-N-pseudoglycosidic linkage. Despite their unique catalytic activity and important role in the biosynthesis of natural products, little is known about the molecular basis governing their substrate specificity and catalysis. Here, we report comparative biochemical and kinetic studies using recombinant OtsA, VldE, and their chimeric proteins with a variety of sugar and pseudosugar substrates. We found that the chimeric enzymes can produce hybrid pseudo-(amino)disaccharides, and an amino group in the acceptor is necessary to facilitate a coupling reaction with a pseudosugar donor. Furthermore, we found that the N-terminal domains of the enzymes not only play a major role in selecting the acceptors, but also control the type of nucleotidyl diphosphate moiety of the donors.
authors.affiliation: Department of Pharmaceutical Sciences, Oregon State University, Corvallis, OR 97331-3507, USA.
authors.forename: Hatem A
authors.initials: HA
authors.lastname: Abuelizz
authors.affiliation: Department of Pharmaceutical Sciences, Oregon State University, Corvallis, OR 97331-3507, USA. Electronic address: taifo.mahmud@oregonstate.edu.
authors.forename: Taifo
authors.initials: T
authors.lastname: Mahmud
mesh_terms: D055162:Biocatalysis; D004926:Escherichia coli / Q000201:enzymology; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016695:Glycosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D048372:Streptomyces coelicolor / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D011993:Recombinant Fusion Proteins; D016695:Glycosyltransferases; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase