3zn2: Difference between revisions
New page: '''Unreleased structure''' The entry 3zn2 is ON HOLD Authors: Schallmey, M., Jekel, P., Tang, L., Majeric-Elenkov, M., Hoeffken, H.W., Hauer, B., Janssen, D.B. Description: protein eng... |
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The | ==protein engineering of halohydrin dehalogenase== | ||
<StructureSection load='3zn2' size='340' side='right'caption='[[3zn2]], [[Resolution|resolution]] 1.80Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[3zn2]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Agrobacterium_tumefaciens Agrobacterium tumefaciens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3ZN2 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3ZN2 FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.8Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=PG5:1-METHOXY-2-[2-(2-METHOXY-ETHOXY]-ETHANE'>PG5</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=3zn2 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3zn2 OCA], [https://pdbe.org/3zn2 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3zn2 RCSB], [https://www.ebi.ac.uk/pdbsum/3zn2 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3zn2 ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/Q93D82_RHIRD Q93D82_RHIRD] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
The cyanide-mediated ring opening of epoxides catalyzed by halohydrin dehalogenases yields beta-hydroxynitriles that are of high interest for synthetic chemistry. The best studied halohydrin dehalogenase to date is the enzyme from Agrobacterium radiobacter, but this enzyme (HheC) exhibits only low cyanolysis activities. Sequence comparison between a pair of related halohydrin dehalogenases from Corynebacterium and Mycobacterium suggested that substitution of a threonine that interacts with the active site might be responsible for the higher cyanolytic activity of the former enzyme. Here we report that a variant of HheC in which this substitution (T134A) is adopted displays an up to 11-fold higher activity in cyanide-mediated epoxide ring-opening. The mutation causes removal of the hydrogen bond between residue 134 and the side chain O of the active site serine 132, which donates a hydrogen bond to the substrate oxygen. The mutation also increases dehalogenase rates with various substrates. Structural analysis revealed that the anion-binding site of the mutant enzyme remained unaltered, showing that the enhanced activity is due to altered interactions with the substrate oxygen rather than changes in the nucleophile binding site. | |||
A single point mutation enhances hydroxynitrile synthesis by halohydrin dehalogenase.,Schallmey M, Jekel P, Tang L, Majeric Elenkov M, Hoffken HW, Hauer B, Janssen DB Enzyme Microb Technol. 2015 Mar;70:50-7. doi: 10.1016/j.enzmictec.2014.12.009., Epub 2014 Dec 24. PMID:25659632<ref>PMID:25659632</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 3zn2" style="background-color:#fffaf0;"></div> | |||
==See Also== | |||
*[[Dehalogenase 3D structures|Dehalogenase 3D structures]] | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Agrobacterium tumefaciens]] | |||
[[Category: Large Structures]] | |||
[[Category: Hauer B]] | |||
[[Category: Hoeffken HW]] | |||
[[Category: Janssen DB]] | |||
[[Category: Jekel P]] | |||
[[Category: Majeric-Elenkov M]] | |||
[[Category: Schallmey M]] | |||
[[Category: Tang L]] | |||
Latest revision as of 14:06, 20 December 2023
protein engineering of halohydrin dehalogenaseprotein engineering of halohydrin dehalogenase
Structural highlights
FunctionPublication Abstract from PubMedThe cyanide-mediated ring opening of epoxides catalyzed by halohydrin dehalogenases yields beta-hydroxynitriles that are of high interest for synthetic chemistry. The best studied halohydrin dehalogenase to date is the enzyme from Agrobacterium radiobacter, but this enzyme (HheC) exhibits only low cyanolysis activities. Sequence comparison between a pair of related halohydrin dehalogenases from Corynebacterium and Mycobacterium suggested that substitution of a threonine that interacts with the active site might be responsible for the higher cyanolytic activity of the former enzyme. Here we report that a variant of HheC in which this substitution (T134A) is adopted displays an up to 11-fold higher activity in cyanide-mediated epoxide ring-opening. The mutation causes removal of the hydrogen bond between residue 134 and the side chain O of the active site serine 132, which donates a hydrogen bond to the substrate oxygen. The mutation also increases dehalogenase rates with various substrates. Structural analysis revealed that the anion-binding site of the mutant enzyme remained unaltered, showing that the enhanced activity is due to altered interactions with the substrate oxygen rather than changes in the nucleophile binding site. A single point mutation enhances hydroxynitrile synthesis by halohydrin dehalogenase.,Schallmey M, Jekel P, Tang L, Majeric Elenkov M, Hoffken HW, Hauer B, Janssen DB Enzyme Microb Technol. 2015 Mar;70:50-7. doi: 10.1016/j.enzmictec.2014.12.009., Epub 2014 Dec 24. PMID:25659632[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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