8xiz
From Proteopedia
(Difference between revisions)
| Line 1: | Line 1: | ||
| - | ==Crystal structure of an epoxide hydrolase mutant A250IC/L344V | + | ==Crystal structure of an epoxide hydrolase mutant A250IC/L344V from Aspergillus usamii E001 at 2.17 Angstroms resolution== |
| - | <StructureSection load='8xiz' size='340' side='right'caption='[[8xiz]]' scene=''> | + | <StructureSection load='8xiz' size='340' side='right'caption='[[8xiz]], [[Resolution|resolution]] 2.17Å' scene=''> |
== Structural highlights == | == Structural highlights == | ||
| - | <table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=8XIZ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=8XIZ FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[8xiz]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Aspergillus_usamii Aspergillus usamii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=8XIZ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=8XIZ FirstGlance]. <br> |
| - | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction</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]] 2.175Å</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=8xiz FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=8xiz OCA], [https://pdbe.org/8xiz PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=8xiz RCSB], [https://www.ebi.ac.uk/pdbsum/8xiz PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=8xiz ProSAT]</span></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=8xiz FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=8xiz OCA], [https://pdbe.org/8xiz PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=8xiz RCSB], [https://www.ebi.ac.uk/pdbsum/8xiz PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=8xiz ProSAT]</span></td></tr> | ||
</table> | </table> | ||
| + | == Function == | ||
| + | [https://www.uniprot.org/uniprot/T2B4K5_ASPUS T2B4K5_ASPUS] | ||
| + | <div style="background-color:#fffaf0;"> | ||
| + | == Publication Abstract from PubMed == | ||
| + | Chiral (S)-o-fluorostyrene oxide (oFSO) and vicinal diol (R)-o-fluorophenylethane-1,2-diol (oFPED) are important intermediates for synthesizing treatments for neuropathic diseases. This study aimed to engineer Aspergillus usamii epoxide hydrolase (AuEH2) through a rational mutagenesis strategy to customize high enantioselectivity mutant for rac-oFSO. Out of 181 single-site mutants screened, six showed elevated enantiomeric ratio (E value) ranging from 32 to 108 according to E value and activity mutability landscapes. By combinatorial mutagenesis of A250I with other five single-site mutants, we constructed five double-site mutants, with the best-performing mutant, D5 (A250I/L344V), achieving an E value of 180. This mutant enabled the efficient kinetic resolution of 400 mM rac-oFSO in pure water system using E. coli/Aueh2(A250I/L344V), yielding (S)-oFSO (>99 % ee(s), 50 % yield(s)) and (R)-oFPED (>99 % ee(p), 50 % yield(p)) with space-time yields (STYs) of 331.5 and 376.1 g/L/d, respectively. Combining crystal structure resolution with theoretical computations clarified the enantioselectivity mechanism of D5, demonstrating that A250I reduced the funnel-shaped substrate binding pocket (SBP) while L344V extended its bottom, enhancing specific recognition of (R)-oFSO and inhibiting (S)-oFSO hydrolysis. These findings provide valuable insights for designing highly enantioselective enzyme mutants, advancing the field of asymmetric synthesis of chiral compounds using green biocatalytic processes. | ||
| + | |||
| + | Rational mutagenesis of an epoxide hydrolase and its structural mechanism for the enantioselectivity improvement toward chiral ortho-fluorostyrene oxide.,Lu ZY, Liao X, Jing WW, Liu KK, Ren QG, He YC, Hu D Int J Biol Macromol. 2024 Dec;282(Pt 3):136864. doi: , 10.1016/j.ijbiomac.2024.136864. Epub 2024 Oct 29. PMID:39476898<ref>PMID:39476898</ref> | ||
| + | |||
| + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
| + | </div> | ||
| + | <div class="pdbe-citations 8xiz" style="background-color:#fffaf0;"></div> | ||
| + | == References == | ||
| + | <references/> | ||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
| + | [[Category: Aspergillus usamii]] | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
| - | [[Category: Hu | + | [[Category: Hu BC]] |
| + | [[Category: Hu D]] | ||
| + | [[Category: Lu ZY]] | ||
| + | [[Category: Tang CD]] | ||
Current revision
Crystal structure of an epoxide hydrolase mutant A250IC/L344V from Aspergillus usamii E001 at 2.17 Angstroms resolution
| |||||||||||
Categories: Aspergillus usamii | Large Structures | Hu BC | Hu D | Lu ZY | Tang CD
