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| | ==D308A, F268V, D469Y, A513V, and Y515S quintuple mutant of Bacillus circulans T-3040 cycloisomaltooligosaccharide glucanotransferase complexed with isomaltoundecaose== | | ==D308A, F268V, D469Y, A513V, and Y515S quintuple mutant of Bacillus circulans T-3040 cycloisomaltooligosaccharide glucanotransferase complexed with isomaltoundecaose== |
| - | <StructureSection load='3wnp' size='340' side='right' caption='[[3wnp]], [[Resolution|resolution]] 2.80Å' scene=''> | + | <StructureSection load='3wnp' size='340' side='right'caption='[[3wnp]], [[Resolution|resolution]] 2.80Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[3wnp]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Atcc_24 Atcc 24]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3WNP OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3WNP FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[3wnp]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Niallia_circulans Niallia circulans]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3WNP OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3WNP FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=BGC:BETA-D-GLUCOSE'>BGC</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=GLC:ALPHA-D-GLUCOSE'>GLC</scene>, <scene name='pdbligand=MES:2-(N-MORPHOLINO)-ETHANESULFONIC+ACID'>MES</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</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]] 2.8Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[3wnk|3wnk]], [[3wnl|3wnl]], [[3wnm|3wnm]], [[3wnn|3wnn]], [[3wno|3wno]]</td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=BGC:BETA-D-GLUCOSE'>BGC</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=GLC:ALPHA-D-GLUCOSE'>GLC</scene>, <scene name='pdbligand=MES:2-(N-MORPHOLINO)-ETHANESULFONIC+ACID'>MES</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">cit ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1397 ATCC 24])</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=3wnp FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3wnp OCA], [https://pdbe.org/3wnp PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3wnp RCSB], [https://www.ebi.ac.uk/pdbsum/3wnp PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3wnp ProSAT]</span></td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Cycloisomaltooligosaccharide_glucanotransferase Cycloisomaltooligosaccharide glucanotransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.248 2.4.1.248] </span></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=3wnp FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3wnp OCA], [http://pdbe.org/3wnp PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=3wnp RCSB], [http://www.ebi.ac.uk/pdbsum/3wnp PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=3wnp ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/CTA1_BACCI CTA1_BACCI]] Produces cycloisomaltooligosaccharide from dextran containing 7, 8 or 9 glucose units. The enzyme is specific for (1->6)-alpha-D-glucans (dextrans) and, without activity toward (1->4)-alpha-D-glucans, such as amylose. It also has no activity on oligosaccharides, such as amylopectin and pullulan, containing (1->6)-alpha-D-glucosidic linkages at branch points.<ref>PMID:7515357</ref> | + | [https://www.uniprot.org/uniprot/CTA1_NIACI CTA1_NIACI] Produces cycloisomaltooligosaccharide from dextran containing 7, 8 or 9 glucose units. The enzyme is specific for (1->6)-alpha-D-glucans (dextrans) and, without activity toward (1->4)-alpha-D-glucans, such as amylose. It also has no activity on oligosaccharides, such as amylopectin and pullulan, containing (1->6)-alpha-D-glucosidic linkages at branch points.<ref>PMID:7515357</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: Atcc 24]] | + | [[Category: Large Structures]] |
| - | [[Category: Cycloisomaltooligosaccharide glucanotransferase]] | + | [[Category: Niallia circulans]] |
| - | [[Category: Fujimoto, Z]] | + | [[Category: Fujimoto Z]] |
| - | [[Category: Funane, K]] | + | [[Category: Funane K]] |
| - | [[Category: Kimura, A]] | + | [[Category: Kimura A]] |
| - | [[Category: Kimura, K]] | + | [[Category: Kimura K]] |
| - | [[Category: Kitamura, S]] | + | [[Category: Kitamura S]] |
| - | [[Category: Momma, M]] | + | [[Category: Momma M]] |
| - | [[Category: Suzuki, N]] | + | [[Category: Suzuki N]] |
| - | [[Category: Suzuki, R]] | + | [[Category: Suzuki R]] |
| - | [[Category: 6-glucan]]
| + | |
| - | [[Category: Alpha-1]]
| + | |
| - | [[Category: Beta-jelly roll]]
| + | |
| - | [[Category: C2 type immunoglobulin fold]]
| + | |
| - | [[Category: Glycoside hydrolase]]
| + | |
| - | [[Category: Greek key]]
| + | |
| - | [[Category: Transferase]]
| + | |
| Structural highlights
3wnp is a 2 chain structure with sequence from Niallia circulans. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
| | Method: | X-ray diffraction, Resolution 2.8Å |
| Ligands: | , , , , , |
| Resources: | FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT |
Function
CTA1_NIACI Produces cycloisomaltooligosaccharide from dextran containing 7, 8 or 9 glucose units. The enzyme is specific for (1->6)-alpha-D-glucans (dextrans) and, without activity toward (1->4)-alpha-D-glucans, such as amylose. It also has no activity on oligosaccharides, such as amylopectin and pullulan, containing (1->6)-alpha-D-glucosidic linkages at branch points.[1]
Publication Abstract from PubMed
Cycloisomaltooligosaccharide glucanotransferase (CITase) is a member of the glycoside hydrolase family 66, and it produces cycloisomaltooligosaccharides (CIs). Small CIs (CI-7-9) and large CIs (CI->/=10) are designated as oligosaccharide-type CIs (oligo-CIs) and megalosaccharide-type CIs (megalo-CIs), respectively. CITase from Bacillus circulans T-3040 (BcCITase) produces mainly CI-8 with little megalo-CIs. It has two family 35 carbohydrate-binding modules (BcCBM35-1 and BcCBM35-2). BcCBM35-1 is inserted in a catalytic domain of BcCITase, and BcCBM35-2 is located at the C-terminal region. Our previous studies suggested that BcCBM35-1 has two substrate-binding sites (B-1 and B-2). We implemented site-directed mutagenesis of BcCITase to explore the preference for product size on the basis of the 3D structure of BcCITase. Mutational studies provided evidence that B-1 and B-2 contribute to recruiting substrate and maintaining product size, respectively. A mutant (mutant-R) with four mutations (F268V, D469Y, A513V and Y515S) produced three times as much megalo-CIs (CI-10-12) and 1.5 times as much total CIs (CI-7-12) as compared with the wild-type BcCITase. The 3D structure of the substrate-enzyme complex of mutant-R suggested that the modified product size specificity was attributable to the construction of novel substrate-binding sites in the B-2 site of BcCBM35-1, and reactivity was improved by mutation on subsite -3 on the catalytic domain.
Molecular engineering of cycloisomaltooligosaccharide glucanotransferase from Bacillus circulans T-3040: structural determinants for the reaction product size and reactivity.,Suzuki R, Suzuki N, Fujimoto Z, Momma M, Kimura K, Kitamura S, Kimura A, Funane K Biochem J. 2015 Feb 4. PMID:25649478[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Oguma T, Tobe K, Kobayashi M. Purification and properties of a novel enzyme from Bacillus spp. T-3040, which catalyzes the conversion of dextran to cyclic isomaltooligosaccharides. FEBS Lett. 1994 May 30;345(2-3):135-8. PMID:7515357
- ↑ Suzuki R, Suzuki N, Fujimoto Z, Momma M, Kimura K, Kitamura S, Kimura A, Funane K. Molecular engineering of cycloisomaltooligosaccharide glucanotransferase from Bacillus circulans T-3040: structural determinants for the reaction product size and reactivity. Biochem J. 2015 Feb 4. PMID:25649478 doi:http://dx.doi.org/10.1042/BJ20140860
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