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==Crystal structure of GH1 beta-glucosidase Td2F2==
==Crystal structure of GH1 beta-glucosidase Td2F2==
<StructureSection load='3wh5' size='340' side='right' caption='[[3wh5]], [[Resolution|resolution]] 1.60&Aring;' scene=''>
<StructureSection load='3wh5' size='340' side='right' caption='[[3wh5]], [[Resolution|resolution]] 1.60&Aring;' scene=''>
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<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[3wh6|3wh6]], [[3wh7|3wh7]], [[3wh8|3wh8]]</td></tr>
<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[3wh6|3wh6]], [[3wh7|3wh7]], [[3wh8|3wh8]]</td></tr>
<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Beta-glucosidase Beta-glucosidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.21 3.2.1.21] </span></td></tr>
<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Beta-glucosidase Beta-glucosidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.21 3.2.1.21] </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=3wh5 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3wh5 OCA], [http://www.rcsb.org/pdb/explore.do?structureId=3wh5 RCSB], [http://www.ebi.ac.uk/pdbsum/3wh5 PDBsum]</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=3wh5 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3wh5 OCA], [http://pdbe.org/3wh5 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=3wh5 RCSB], [http://www.ebi.ac.uk/pdbsum/3wh5 PDBsum]</span></td></tr>
</table>
</table>
<div style="background-color:#fffaf0;">
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
== Publication Abstract from PubMed ==
The beta-glucosidase encoded by the td2f2 gene was isolated from a compost microbial metagenomic library by functional screening. The protein was identified to be a member of the glycoside hydrolase family 1 and was overexpressed in Escherichia coli, purified, and biochemically characterized. The recombinant beta-glucosidase, Td2F2, exhibited enzymatic activity with beta-glycosidic substrates, with preferences for glucose, fucose, and galactose. Hydrolysis occurred at the nonreducing end and in an exo manner. The order of catalytic efficiency for glucodisaccharides and cellooligosaccharides was sophorose &gt; cellotetraose &gt; cellotriose &gt; laminaribiose &gt; cellobiose &gt; cellopentaose &gt; gentiobiose, respectively. Intriguingly, the p-nitrophenyl-beta-D-glucopyranoside hydrolysis activity of Td2F2 was activated by various monosaccharides and sugar alcohols. At a D-glucose concentration of 1000 mM, enzyme activity was 6.7-fold higher than that observed in the absence of D-glucose. With 31.3 mM D-glucose, Td2F2 catalyzed transglycosylation to generate sophorose, laminaribiose, cellobiose, and gentiobiose. Transglycosylation products were detected under all activated conditions, suggesting that the activity enhancement induced by monosaccharides and sugar alcohols may be due to the transglycosylation activity of the enzyme. These results show that Td2F2 obtained from a compost microbial metagenome may be a potent candidate for industrial applications.
beta-Glucosidase Td2F2 isolated from a compost metagenome has high glucose tolerance and transglycosylation activity. In this study, we determined the high-resolution crystal structure of Td2F2. It has a unique structure at the -1 subsite that is important for substrate specificity but not for glucose tolerance. To elucidate the mechanism(s) of glucose tolerance, we isolated a glucose-sensitive Td2F2 mutant using random mutagenesis. In this mutant, Asn223 residue located between subsites +1 and +2 was mutated. The Asn223 mutation resulted in reduced glucose tolerance and transglycosylation activity, and drastically changed substrate specificity. These results indicate that the structure between subsites +1 and +2 is critical for the glucose tolerance and substrate specificity of Td2F2. Our findings shed light on the glucose tolerance and transglycosylation activity mechanisms of glycoside hydrolase family 1 beta-glucosidases. This article is protected by copyright. All rights reserved.


Characterization of a novel beta-glucosidase from a compost microbial metagenome with strong transglycosylation activity.,Uchiyama T, Miyazaki K, Yaoi K J Biol Chem. 2013 Jun 21;288(25):18325-34. doi: 10.1074/jbc.M113.471342. Epub, 2013 May 9. PMID:23661705<ref>PMID:23661705</ref>
Crystal structure and identification of a key amino acid for glucose tolerance, substrate specificity and transglycosylation activity of metagenomic beta-glucosidase Td2F2.,Matsuzawa T, Jo T, Uchiyama T, Manninen JA, Arakawa T, Miyazaki K, Fushinobu S, Yaoi K FEBS J. 2016 Apr 19. doi: 10.1111/febs.13743. PMID:27092463<ref>PMID:27092463</ref>


From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
</div>
<div class="pdbe-citations 3wh5" style="background-color:#fffaf0;"></div>
==See Also==
*[[Beta-glucosidase|Beta-glucosidase]]
== References ==
== References ==
<references/>
<references/>

Revision as of 19:29, 10 May 2016

Crystal structure of GH1 beta-glucosidase Td2F2Crystal structure of GH1 beta-glucosidase Td2F2

Structural highlights

3wh5 is a 1 chain structure. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Ligands:, ,
Activity:Beta-glucosidase, with EC number 3.2.1.21
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum

Publication Abstract from PubMed

beta-Glucosidase Td2F2 isolated from a compost metagenome has high glucose tolerance and transglycosylation activity. In this study, we determined the high-resolution crystal structure of Td2F2. It has a unique structure at the -1 subsite that is important for substrate specificity but not for glucose tolerance. To elucidate the mechanism(s) of glucose tolerance, we isolated a glucose-sensitive Td2F2 mutant using random mutagenesis. In this mutant, Asn223 residue located between subsites +1 and +2 was mutated. The Asn223 mutation resulted in reduced glucose tolerance and transglycosylation activity, and drastically changed substrate specificity. These results indicate that the structure between subsites +1 and +2 is critical for the glucose tolerance and substrate specificity of Td2F2. Our findings shed light on the glucose tolerance and transglycosylation activity mechanisms of glycoside hydrolase family 1 beta-glucosidases. This article is protected by copyright. All rights reserved.

Crystal structure and identification of a key amino acid for glucose tolerance, substrate specificity and transglycosylation activity of metagenomic beta-glucosidase Td2F2.,Matsuzawa T, Jo T, Uchiyama T, Manninen JA, Arakawa T, Miyazaki K, Fushinobu S, Yaoi K FEBS J. 2016 Apr 19. doi: 10.1111/febs.13743. PMID:27092463[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

See Also

References

  1. Matsuzawa T, Jo T, Uchiyama T, Manninen JA, Arakawa T, Miyazaki K, Fushinobu S, Yaoi K. Crystal structure and identification of a key amino acid for glucose tolerance, substrate specificity and transglycosylation activity of metagenomic beta-glucosidase Td2F2. FEBS J. 2016 Apr 19. doi: 10.1111/febs.13743. PMID:27092463 doi:http://dx.doi.org/10.1111/febs.13743

3wh5, resolution 1.60Å

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