8wg2 | pdb_00008wg2

Crystal structure of GH97 glucodextranase mutant E509Q from Flavobacterium johnsoniae in complex with isomaltotriose

spinqualitylabelsall models 8wg2, resolution 2.45Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 8wg2 is a 4 chain structure with sequence from Flavobacterium johnsoniae UW101. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 2.45Å Ligands: BGC, CA, GLC Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function A5FBI0_FLAJ1 Publication Abstract from PubMed Glycoside hydrolase family 97 (GH97) comprises enzymes like anomer-inverting alpha-glucoside hydrolases (i.e., glucoamylase) and anomer-retaining alpha-galactosidases. In a soil bacterium, Flavobacterium johnsoniae, we previously identified a GH97 enzyme (FjGH97A) within the branched dextran utilization locus. It functions as an alpha-glucoside hydrolase, targeting alpha-(1-->6)-glucosidic linkages in dextran and isomaltooligosaccharides (i.e., glucodextranase). FjGH97A exhibits a preference for alpha-(1-->6)-glucoside linkages over alpha-(1-->4)-linkages, while Bacteroides thetaiotaomicron glucoamylase SusB (with 69% sequence identity), which is involved in the starch utilization system, exhibits the highest specificity for alpha-(1-->4)-glucosidic linkages. Here, we examined the crystal structures of FjGH97A in complexes with glucose, panose, or isomaltotriose, and analyzed the substrate preferences of its mutants to identify the amino acid residues that determine the substrate specificity for alpha-(1-->4)- and alpha-(1-->6)-glucosidic linkages. The overall structure of FjGH97A resembles other GH97 enzymes, with conserved catalytic residues similar to anomer-inverting GH97 enzymes. A comparison of active sites between FjGH97A and SusB revealed differences in amino acid residues at subsites +1 and +2 (specifically Ala195 and Ile378 in FjGH97A). Among the three mutants (A195S, I378F, and A195S-I378F), A195S and A195S-I378F exhibited increased activity toward alpha-(1-->4)-glucoside bonds compared to alpha-(1-->6)-glucoside bonds. This suggests that Ala195, located on the Gly184-Thr203 loop (named loop-N) conserved within the GH97 subgroup, including FjGH97A and SusB, holds significance in determining linkage specificity. The conservation of alanine in the active site of the GH97 enzymes, within the same gene cluster as the putative dextranase, indicates its crucial role in determining the specificity for alpha-(1-->6)-glucoside linkage. Structural insights into alpha-(1-->6)-linkage preference of GH97 glucodextranase from Flavobacterium johnsoniae.,Nakamura S, Kurata R, Miyazaki T FEBS J. 2024 Apr 25. doi: 10.1111/febs.17139. PMID:38661728 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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