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| | ==Structure of a 37-fold mutant of halohydrin dehalogenase (HheC) with chloride bound== | | ==Structure of a 37-fold mutant of halohydrin dehalogenase (HheC) with chloride bound== |
| - | <StructureSection load='4iy1' size='340' side='right' caption='[[4iy1]], [[Resolution|resolution]] 2.10Å' scene=''> | + | <StructureSection load='4iy1' size='340' side='right'caption='[[4iy1]], [[Resolution|resolution]] 2.10Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4iy1]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/"achromobacter_radiobacter"_(beijerinck_and_van_delden_1902)_bergey_et_al._1934 "achromobacter radiobacter" (beijerinck and van delden 1902) bergey et al. 1934]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4IY1 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4IY1 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4iy1]] 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=4IY1 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4IY1 FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</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]] 2.1Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4ixt|4ixt]], [[4ixw|4ixw]]</td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">hheC ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=358 "Achromobacter radiobacter" (Beijerinck and van Delden 1902) Bergey et al. 1934])</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=4iy1 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4iy1 OCA], [https://pdbe.org/4iy1 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4iy1 RCSB], [https://www.ebi.ac.uk/pdbsum/4iy1 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4iy1 ProSAT]</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=4iy1 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4iy1 OCA], [http://pdbe.org/4iy1 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4iy1 RCSB], [http://www.ebi.ac.uk/pdbsum/4iy1 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4iy1 ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/Q93D82_RHIRD Q93D82_RHIRD] |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | </div> | | </div> |
| | <div class="pdbe-citations 4iy1" style="background-color:#fffaf0;"></div> | | <div class="pdbe-citations 4iy1" style="background-color:#fffaf0;"></div> |
| | + | |
| | + | ==See Also== |
| | + | *[[Dehalogenase 3D structures|Dehalogenase 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Breuer, M]] | + | [[Category: Agrobacterium tumefaciens]] |
| - | [[Category: Dijkstra, B W]] | + | [[Category: Large Structures]] |
| - | [[Category: Floor, R J]] | + | [[Category: Breuer M]] |
| - | [[Category: Hauer, B]]
| + | [[Category: Dijkstra BW]] |
| - | [[Category: Janssen, D B]]
| + | [[Category: Floor RJ]] |
| - | [[Category: Jekel, P A]]
| + | [[Category: Hauer B]] |
| - | [[Category: Schallmey, M]]
| + | [[Category: Janssen DB]] |
| - | [[Category: Wijma, H J]] | + | [[Category: Jekel PA]] |
| - | [[Category: Atorvastatin synthesis]] | + | [[Category: Schallmey M]] |
| - | [[Category: Backbone change]] | + | [[Category: Wijma HJ]] |
| - | [[Category: Coupled mutation]] | + | |
| - | [[Category: Cyanolysis]] | + | |
| - | [[Category: Dehalogenase]] | + | |
| - | [[Category: Directed evolution]] | + | |
| - | [[Category: Enantioselectivity change]]
| + | |
| - | [[Category: Lyase]]
| + | |
| - | [[Category: Proline-induced]]
| + | |
| - | [[Category: Protein engineering]]
| + | |
| - | [[Category: Short-chain dehydrogenase/reductase enzyme superfamily]]
| + | |
| - | [[Category: Synergistic mutation]]
| + | |
| - | [[Category: Thermostability]]
| + | |
| Structural highlights
Function
Q93D82_RHIRD
Publication Abstract from PubMed
Two highly engineered halohydrin dehalogenase variants were characterized in terms of their performance in dehalogenation and epoxide cyanolysis reactions. Both enzyme variants outperformed the wild-type enzyme in the cyanolysis of ethyl (S)-3,4-epoxybutyrate, a conversion yielding ethyl (R)-4-cyano-3-hydroxybutyrate, an important chiral building block for statin synthesis. One of the enzyme variants, HheC2360, displayed catalytic rates for this cyanolysis reaction enhanced up to tenfold. Furthermore, the enantioselectivity of this variant was the opposite of that of the wild-type enzyme, both for dehalogenation and for cyanolysis reactions. The 37-fold mutant HheC2360 showed an increase in thermal stability of 8 degrees C relative to the wild-type enzyme. Crystal structures of this enzyme were elucidated with chloride and ethyl (S)-3,4-epoxybutyrate or with ethyl (R)-4-cyano-3-hydroxybutyrate bound in the active site. The observed increase in temperature stability was explained in terms of a substantial increase in buried surface area relative to the wild-type HheC, together with enhanced interfacial interactions between the subunits that form the tetramer. The structures also revealed that the substrate binding pocket was modified both by substitutions and by backbone movements in loops surrounding the active site. The observed changes in the mutant structures are partly governed by coupled mutations, some of which are necessary to remove steric clashes or to allow backbone movements to occur. The importance of interactions between substitutions suggests that efficient directed evolution strategies should allow for compensating and synergistic mutations during library design.
Biocatalytic and structural properties of a highly engineered halohydrin dehalogenase.,Schallmey M, Floor RJ, Hauer B, Breuer M, Jekel PA, Wijma HJ, Dijkstra BW, Janssen DB Chembiochem. 2013 May 10;14(7):870-81. doi: 10.1002/cbic.201300005. Epub 2013 Apr, 12. PMID:23585096[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Schallmey M, Floor RJ, Hauer B, Breuer M, Jekel PA, Wijma HJ, Dijkstra BW, Janssen DB. Biocatalytic and structural properties of a highly engineered halohydrin dehalogenase. Chembiochem. 2013 May 10;14(7):870-81. doi: 10.1002/cbic.201300005. Epub 2013 Apr, 12. PMID:23585096 doi:10.1002/cbic.201300005
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