5h4r: Difference between revisions
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==the complex of Glycoside Hydrolase 5 Lichenase from Caldicellulosiruptor sp. F32 E188Q mutant and cellotetraose== | |||
<StructureSection load='5h4r' size='340' side='right' caption='[[5h4r]], [[Resolution|resolution]] 1.70Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[5h4r]] is a 1 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5H4R OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5H4R FirstGlance]. <br> | |||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=CTT:BETA-D-GLUCOPYRANOSYL-(1- 4)-BETA-D-GLUCOPYRANOSYL-(1- 4)-BETA-D-GLUCOPYRANOSYL-(1- 4)-BETA-D-GLUCOPYRANOSE'>CTT</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene></td></tr> | |||
<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4x0v|4x0v]]</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=5h4r FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5h4r OCA], [http://pdbe.org/5h4r PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5h4r RCSB], [http://www.ebi.ac.uk/pdbsum/5h4r PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5h4r ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Glycoside hydrolase (GH) family 5 is one of the largest GH families with various GH activities including lichenase, but the structural basis of the GH5 lichenase activity is still unknown. A novel thermostable lichenase F32EG5 belonging to GH5 was identified from an extremely thermophilic bacterium Caldicellulosiruptor sp. F32. F32EG5 is a bi-functional cellulose and lichenan-degrading enzyme and exhibited a high activity on beta-1,3-1,4-glucan but side activity on cellulose. Thin-layer chromatography and NMR analyses indicated that F32EG5 cleaved the beta-1,4 linkage or the beta-1,3 linkage while a 4- O -substitued glucose residue linked to a glucose residue through a beta-1,3 linkage, which is completely different from extensively studied GH16 lichenase that catalyses strict endo-hydrolysis of the beta-1,4-glycosidic linkage adjacent to a 3- O -substitued glucose residue in the mixed linked beta-glucans. The crystal structure of F32EG5 was determined to 2.8 A resolution and the crystal structure of the complex of F32EG5 E193Q mutant and cellotetraose was determined to 1.7 A resolution, which revealed that the exit subsites of substrate binding sites contribute to both thermostability and substrate specificity of F32EG5. The sugar chain showed a sharp bend in the complex structure, suggesting that a substrate cleft fitting to the bent sugar chains in lichenan is a common feature of GH5 lichenases. The mechanism of thermostability and substrate selectivity of F32EG5 was further demonstrated by molecular dynamics simulation and site-directed mutagenesis. These results provide biochemical and structural insight into thermostability and substrate selectivity of GH5 lichenases which have potential in industrial processes. | |||
Structural Insights into the Substrate Specificity of a Glycoside Hydrolase Family 5 Lichenase from Caldicellulosiruptor sp. F32.,Meng DD, Liu X, Dong S, Wang YF, Ma XQ, Zhou H, Wang X, Yao LS, Feng Y, Li FL Biochem J. 2017 Aug 24. pii: BCJ20170328. doi: 10.1042/BCJ20170328. PMID:28838949<ref>PMID:28838949</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 5h4r" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Dong, S]] | [[Category: Dong, S]] | ||
[[Category: | [[Category: Feng, Y]] | ||
[[Category: Liu, X]] | [[Category: Liu, X]] | ||
[[Category: Wang, X]] | [[Category: Wang, X]] | ||
[[Category: | [[Category: Zhou, H]] | ||
[[Category: Glycoside hydrolase 5 lichenase]] | |||
[[Category: Hydrolase]] | |||
Revision as of 13:26, 13 September 2017
the complex of Glycoside Hydrolase 5 Lichenase from Caldicellulosiruptor sp. F32 E188Q mutant and cellotetraosethe complex of Glycoside Hydrolase 5 Lichenase from Caldicellulosiruptor sp. F32 E188Q mutant and cellotetraose
Structural highlights
Publication Abstract from PubMedGlycoside hydrolase (GH) family 5 is one of the largest GH families with various GH activities including lichenase, but the structural basis of the GH5 lichenase activity is still unknown. A novel thermostable lichenase F32EG5 belonging to GH5 was identified from an extremely thermophilic bacterium Caldicellulosiruptor sp. F32. F32EG5 is a bi-functional cellulose and lichenan-degrading enzyme and exhibited a high activity on beta-1,3-1,4-glucan but side activity on cellulose. Thin-layer chromatography and NMR analyses indicated that F32EG5 cleaved the beta-1,4 linkage or the beta-1,3 linkage while a 4- O -substitued glucose residue linked to a glucose residue through a beta-1,3 linkage, which is completely different from extensively studied GH16 lichenase that catalyses strict endo-hydrolysis of the beta-1,4-glycosidic linkage adjacent to a 3- O -substitued glucose residue in the mixed linked beta-glucans. The crystal structure of F32EG5 was determined to 2.8 A resolution and the crystal structure of the complex of F32EG5 E193Q mutant and cellotetraose was determined to 1.7 A resolution, which revealed that the exit subsites of substrate binding sites contribute to both thermostability and substrate specificity of F32EG5. The sugar chain showed a sharp bend in the complex structure, suggesting that a substrate cleft fitting to the bent sugar chains in lichenan is a common feature of GH5 lichenases. The mechanism of thermostability and substrate selectivity of F32EG5 was further demonstrated by molecular dynamics simulation and site-directed mutagenesis. These results provide biochemical and structural insight into thermostability and substrate selectivity of GH5 lichenases which have potential in industrial processes. Structural Insights into the Substrate Specificity of a Glycoside Hydrolase Family 5 Lichenase from Caldicellulosiruptor sp. F32.,Meng DD, Liu X, Dong S, Wang YF, Ma XQ, Zhou H, Wang X, Yao LS, Feng Y, Li FL Biochem J. 2017 Aug 24. pii: BCJ20170328. doi: 10.1042/BCJ20170328. PMID:28838949[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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