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| | ==Crystal structure of Neisseria polysaccharea amylosucrase mutant efficient for the synthesis of controlled size maltooligosaccharides== | | ==Crystal structure of Neisseria polysaccharea amylosucrase mutant efficient for the synthesis of controlled size maltooligosaccharides== |
| - | <StructureSection load='5n7j' size='340' side='right' caption='[[5n7j]], [[Resolution|resolution]] 2.00Å' scene=''> | + | <StructureSection load='5n7j' size='340' side='right'caption='[[5n7j]], [[Resolution|resolution]] 2.00Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5n7j]] is a 1 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5N7J OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5N7J FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5n7j]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Neisseria_polysaccharea Neisseria polysaccharea]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5N7J OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5N7J FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=1PE:PENTAETHYLENE+GLYCOL'>1PE</scene>, <scene name='pdbligand=MLR:MALTOTRIOSE'>MLR</scene>, <scene name='pdbligand=PG4:TETRAETHYLENE+GLYCOL'>PG4</scene>, <scene name='pdbligand=SUC:SUCROSE'>SUC</scene>, <scene name='pdbligand=TRS:2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL'>TRS</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.001Å</td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Amylosucrase Amylosucrase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.4 2.4.1.4] </span></td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=1PE:PENTAETHYLENE+GLYCOL'>1PE</scene>, <scene name='pdbligand=FRU:FRUCTOSE'>FRU</scene>, <scene name='pdbligand=GLC:ALPHA-D-GLUCOSE'>GLC</scene>, <scene name='pdbligand=PG4:TETRAETHYLENE+GLYCOL'>PG4</scene>, <scene name='pdbligand=PRD_900003:sucrose'>PRD_900003</scene>, <scene name='pdbligand=PRD_900009:alpha-maltotriose'>PRD_900009</scene>, <scene name='pdbligand=TRS:2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL'>TRS</scene></td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5n7j FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5n7j OCA], [http://pdbe.org/5n7j PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5n7j RCSB], [http://www.ebi.ac.uk/pdbsum/5n7j PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5n7j ProSAT]</span></td></tr> | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5n7j FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5n7j OCA], [https://pdbe.org/5n7j PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5n7j RCSB], [https://www.ebi.ac.uk/pdbsum/5n7j PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5n7j ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/AMYS_NEIPO AMYS_NEIPO]] Catalyzes the synthesis of alpha-glucan from sucrose. Catalyzes, in addition, sucrose hydrolysis, maltose and maltotriose synthesis by successive transfers of the glucosyl moiety of sucrose onto the released glucose, and finally turanose and trehalulose synthesis, these two sucrose isomers being obtained by glucosyl transfer onto fructose.<ref>PMID:9882648</ref> | + | [https://www.uniprot.org/uniprot/AMYS_NEIPO AMYS_NEIPO] Catalyzes the synthesis of alpha-glucan from sucrose. Catalyzes, in addition, sucrose hydrolysis, maltose and maltotriose synthesis by successive transfers of the glucosyl moiety of sucrose onto the released glucose, and finally turanose and trehalulose synthesis, these two sucrose isomers being obtained by glucosyl transfer onto fructose.<ref>PMID:9882648</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Amylosucrase]] | + | [[Category: Large Structures]] |
| - | [[Category: Tranier, S]]
| + | |
| - | [[Category: Verges, A]]
| + | |
| - | [[Category: Gh13]]
| + | |
| - | [[Category: Glucosyl transferase]]
| + | |
| - | [[Category: Glycoside hydrolase]]
| + | |
| - | [[Category: Hydrolase]]
| + | |
| - | [[Category: Mutant]]
| + | |
| | [[Category: Neisseria polysaccharea]] | | [[Category: Neisseria polysaccharea]] |
| | + | [[Category: Tranier S]] |
| | + | [[Category: Verges A]] |
| Structural highlights
5n7j is a 1 chain structure with sequence from Neisseria polysaccharea. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
| | Method: | X-ray diffraction, Resolution 2.001Å |
| Ligands: | , , , , , , |
| Resources: | FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT |
Function
AMYS_NEIPO Catalyzes the synthesis of alpha-glucan from sucrose. Catalyzes, in addition, sucrose hydrolysis, maltose and maltotriose synthesis by successive transfers of the glucosyl moiety of sucrose onto the released glucose, and finally turanose and trehalulose synthesis, these two sucrose isomers being obtained by glucosyl transfer onto fructose.[1]
Publication Abstract from PubMed
Amylosucrase from Neisseria polysaccharea naturally catalyzes the synthesis of alpha-1,4 glucans from sucrose. The product profile is quite polydisperse, ranging from soluble chains called maltooligosaccharides to high-molecular weight insoluble amylose. This enzyme was recently subjected to engineering of its active site to enable recognition of non-natural acceptor substrates. Libraries of variants were constructed and screened on sucrose, allowing the identification of a mutant that showed a 6-fold enhanced activity toward sucrose compared to the wild-type enzyme. Furthermore, its product profile was unprecedented, as only soluble maltooligosaccharides of controlled size chains (2<DP<21) with a narrow polydispersity were observed. This variant, containing 9 mutations in the active site, was characterized at both biochemical and structural levels. Its x-ray structure was determined and further investigated by molecular dynamics to understand the molecular origins of its higher activity on sucrose and higher production of small maltooligosaccharides, with a totally abolished insoluble glucan synthesis.
Engineering of anp efficient mutant of Neisseria polysaccharea amylosucrase for the synthesis of controlled size maltooligosaccharides.,Verges A, Barbe S, Cambon E, Moulis C, Tranier S, Remaud-Simeon M, Andre I Carbohydr Polym. 2017 Oct 1;173:403-411. doi: 10.1016/j.carbpol.2017.06.011. Epub, 2017 Jun 7. PMID:28732882[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ De Montalk GP, Remaud-Simeon M, Willemot RM, Planchot V, Monsan P. Sequence analysis of the gene encoding amylosucrase from Neisseria polysaccharea and characterization of the recombinant enzyme. J Bacteriol. 1999 Jan;181(2):375-81. PMID:9882648
- ↑ Verges A, Barbe S, Cambon E, Moulis C, Tranier S, Remaud-Simeon M, Andre I. Engineering of anp efficient mutant of Neisseria polysaccharea amylosucrase for the synthesis of controlled size maltooligosaccharides. Carbohydr Polym. 2017 Oct 1;173:403-411. doi: 10.1016/j.carbpol.2017.06.011. Epub, 2017 Jun 7. PMID:28732882 doi:http://dx.doi.org/10.1016/j.carbpol.2017.06.011
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