6qps

Structural characterization of a mannuronic acid specific polysaccharide family 6 lyase enzyme from human gut microbiota

spinqualitylabelsall models 6qps, resolution 1.29Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 6qps is a 2 chain structure with sequence from Bacteroides cellulosilyticus. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.287Å Ligands: ACT, CA Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function E2NM07_9BACE Publication Abstract from PubMed Alginate is a linear polysaccharide from brown algae consisting of 1,4-linked beta-d-mannuronic acid (M) and alpha-l-guluronic acid (G) arranged in M, G, and mixed MG blocks. Alginate was assumed to be indigestible in humans, but bacteria isolated from fecal samples can utilize alginate. Moreover, genomes of some human gut microbiome-associated bacteria encode putative alginate-degrading enzymes. Here, we genome-mined a polysaccharide lyase family 6 alginate lyase from the gut bacterium Bacteroides cellulosilyticus (BcelPL6). The structure of recombinant BcelPL6 was solved by X-ray crystallography to 1.3 A resolution, revealing a single-domain, monomeric parallel beta-helix containing a 10-step asparagine ladder characteristic of alginate-converting parallel beta-helix enzymes. Substitutions of the conserved catalytic site residues Lys-249, Arg-270, and His-271 resulted in activity loss. However, imidazole restored the activity of BcelPL6-H271N to 2.5% that of the native enzyme. Molecular docking oriented tetra-mannuronic acid for syn attack correlated with M specificity. Using biochemical analyses, we found that BcelPL6 initially releases unsaturated oligosaccharides of a degree of polymerization of 2-7 from alginate and polyM, which were further degraded to di- and trisaccharides. Unlike other PL6 members, BcelPL6 had low activity on polyMG and none on polyG. Surprisingly, polyG increased BcelPL6 activity on alginate 7-fold. LC-electrospray ionization-MS quantification of products and lack of activity on NaBH4-reduced octa-mannuronic acid indicated that BcelPL6 is an endolyase that further degrades the oligosaccharide products with an intact reducing end. We anticipate that our results advance predictions of the specificity and mode of action of PL6 enzymes. Structural and functional aspects of mannuronic acid-specific PL6 alginate lyase from the human gut microbe Bacteroides cellulosilyticus.,Stender EGP, Dybdahl Andersen C, Fredslund F, Holck J, Solberg A, Teze D, Peters GHJ, Christensen BE, Aachmann FL, Welner DH, Svensson B J Biol Chem. 2019 Nov 22;294(47):17915-17930. doi: 10.1074/jbc.RA119.010206. Epub, 2019 Sep 17. PMID:31530640 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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