8ihq: Difference between revisions
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The | ==Cryo-EM structure of ochratoxin A-detoxifying amidohydrolase ADH3== | ||
<StructureSection load='8ihq' size='340' side='right'caption='[[8ihq]], [[Resolution|resolution]] 2.71Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[8ihq]] is a 8 chain structure with sequence from [https://en.wikipedia.org/wiki/Stenotrophomonas_acidaminiphila Stenotrophomonas acidaminiphila]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=8IHQ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=8IHQ FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Electron Microscopy, [[Resolution|Resolution]] 2.71Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=KCX:LYSINE+NZ-CARBOXYLIC+ACID'>KCX</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></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=8ihq FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=8ihq OCA], [https://pdbe.org/8ihq PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=8ihq RCSB], [https://www.ebi.ac.uk/pdbsum/8ihq PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=8ihq ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/A0A7L8TXW5_9GAMM A0A7L8TXW5_9GAMM] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Ochratoxin A (OTA) is among the most prevalent mycotoxins detected in agroproducts, posing serious threats to human and livestock health. Using enzymes to conduct OTA detoxification is an appealing potential strategy. The recently identified amidohydrolase from Stenotrophomonas acidaminiphila, termed ADH3, is the most efficient OTA-detoxifying enzyme reported thus far and can hydrolyze OTA to nontoxic ochratoxin alpha (OTalpha) and L-beta-phenylalanine (Phe). To elucidate the catalytic mechanism of ADH3, we solved the single-particle cryo-electron microscopy (cryo-EM) structures of apo-form, Phe- and OTA-bound ADH3 to an overall resolution of 2.5-2.7 A. The role of OTA-binding residues was investigated by structural, mutagenesis and biochemical analyses. We also rationally engineered ADH3 and obtained variant S88E, whose catalytic activity was elevated by 3.7-fold. Structural analysis of variant S88E indicates that the E88 side chain provides additional hydrogen bond interactions to the OTalpha moiety. Furthermore, the OTA-hydrolytic activity of variant S88E expressed in Pichia pastoris is comparable to that of Escherichia coli-expressed enzyme, revealing the feasibility of employing the industrial yeast strain to produce ADH3 and its variants for further applications. These results unveil a wealth of information about the catalytic mechanism of ADH3-mediated OTA degradation and provide a blueprint for rational engineering of high-efficiency OTA-detoxifying machineries. | |||
Cryo-EM structure and rational engineering of a superefficient ochratoxin A-detoxifying amidohydrolase.,Dai L, Niu D, Huang JW, Li X, Shen P, Li H, Xie Z, Min J, Hu Y, Yang Y, Guo RT, Chen CC J Hazard Mater. 2023 Sep 15;458:131836. doi: 10.1016/j.jhazmat.2023.131836. Epub , 2023 Jun 12. PMID:37331057<ref>PMID:37331057</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
<div class="pdbe-citations 8ihq" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Stenotrophomonas acidaminiphila]] | |||
[[Category: Chen C-C]] | |||
[[Category: Dai LH]] | |||
[[Category: Guo R-T]] | |||
[[Category: Hu YM]] | |||
[[Category: Huang J-W]] | |||
[[Category: Li H]] | |||
[[Category: Li X]] | |||
[[Category: Niu D]] | |||
[[Category: Shen PP]] | |||
[[Category: Yang Y]] | |||
Latest revision as of 12:12, 30 August 2023
Cryo-EM structure of ochratoxin A-detoxifying amidohydrolase ADH3Cryo-EM structure of ochratoxin A-detoxifying amidohydrolase ADH3
Structural highlights
FunctionPublication Abstract from PubMedOchratoxin A (OTA) is among the most prevalent mycotoxins detected in agroproducts, posing serious threats to human and livestock health. Using enzymes to conduct OTA detoxification is an appealing potential strategy. The recently identified amidohydrolase from Stenotrophomonas acidaminiphila, termed ADH3, is the most efficient OTA-detoxifying enzyme reported thus far and can hydrolyze OTA to nontoxic ochratoxin alpha (OTalpha) and L-beta-phenylalanine (Phe). To elucidate the catalytic mechanism of ADH3, we solved the single-particle cryo-electron microscopy (cryo-EM) structures of apo-form, Phe- and OTA-bound ADH3 to an overall resolution of 2.5-2.7 A. The role of OTA-binding residues was investigated by structural, mutagenesis and biochemical analyses. We also rationally engineered ADH3 and obtained variant S88E, whose catalytic activity was elevated by 3.7-fold. Structural analysis of variant S88E indicates that the E88 side chain provides additional hydrogen bond interactions to the OTalpha moiety. Furthermore, the OTA-hydrolytic activity of variant S88E expressed in Pichia pastoris is comparable to that of Escherichia coli-expressed enzyme, revealing the feasibility of employing the industrial yeast strain to produce ADH3 and its variants for further applications. These results unveil a wealth of information about the catalytic mechanism of ADH3-mediated OTA degradation and provide a blueprint for rational engineering of high-efficiency OTA-detoxifying machineries. Cryo-EM structure and rational engineering of a superefficient ochratoxin A-detoxifying amidohydrolase.,Dai L, Niu D, Huang JW, Li X, Shen P, Li H, Xie Z, Min J, Hu Y, Yang Y, Guo RT, Chen CC J Hazard Mater. 2023 Sep 15;458:131836. doi: 10.1016/j.jhazmat.2023.131836. Epub , 2023 Jun 12. PMID:37331057[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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