7x87
The complex structure of beta-1,2-glucosyltransferase from Ignavibacterium album with sophotetraose observed as sophorose
Structural highlights
FunctionPublication Abstract from PubMedThe IALB_1185 protein, which is encoded in the gene cluster for endo-beta-1,2-glucanase homologs in the genome of Ignavibacterium album, is a glycoside hydrolase family (GH) 35 protein. However, most known GH35 enzymes are beta-galactosidases, which is inconsistent with the components of this gene cluster. Thus, IALB_1185 is expected to possess novel enzymatic properties. Here, we showed using recombinant IALB_1185 that this protein has glycosyltransferase activity toward beta-1,2-glucooligosaccharides, and that the kinetic parameters for beta-1,2-glucooligosaccharides are not within the ranges for general GH enzymes. When various aryl- and alkyl-glucosides were used as acceptors, glycosyltransfer products derived from these acceptors were subsequently detected. Kinetic analysis further revealed that the enzyme has wide aglycone specificity regardless of the anomer, and that the beta-1,2-linked glucose dimer sophorose is an appropriate donor. In the complex of wild-type IALB_1185 with sophorose, the electron density of sophorose was clearly observed at subsites -1 and +1, whereas in the E343Q mutant-sophorose complex, the electron density of sophorose was clearly observed at subsites +1 and +2. This observation suggests that binding at subsites -1 and +2 competes through Glu102, which is consistent with the preference for sophorose as a donor and unsuitability of beta-1,2-glucooligosaccharides as acceptors. A pliable hydrophobic pocket that can accommodate various aglycone moieties was also observed in the complex structures with various glucosides. Overall, our biochemical and structural data are indicative of a novel enzymatic reaction. We propose that IALB_1185 be redefined beta-1,2-glucooligosaccharide:d-glucoside beta-d-glucosyltransferase as a systematic name and beta-1,2-glucosyltransferase as an accepted name. Characterization and structural analyses of a novel glycosyltransferase acting on the beta-1,2-glucosidic linkages.,Kobayashi K, Shimizu H, Tanaka N, Kuramochi K, Nakai H, Nakajima M, Taguchi H J Biol Chem. 2022 Jan 19;298(3):101606. doi: 10.1016/j.jbc.2022.101606. PMID:35065074[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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