4u2f: Difference between revisions
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<StructureSection load='4u2f' size='340' side='right'caption='[[4u2f]], [[Resolution|resolution]] 1.80Å' scene=''> | <StructureSection load='4u2f' size='340' side='right'caption='[[4u2f]], [[Resolution|resolution]] 1.80Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[4u2f]] is a 1 chain structure with sequence from [ | <table><tr><td colspan='2'>[[4u2f]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_knackmussii Pseudomonas knackmussii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4U2F OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4U2F FirstGlance]. <br> | ||
</td></tr><tr id=' | </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=SO4:SULFATE+ION'>SO4</scene></td></tr> | |||
<tr id=' | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=4u2f FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4u2f OCA], [https://pdbe.org/4u2f PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4u2f RCSB], [https://www.ebi.ac.uk/pdbsum/4u2f PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4u2f ProSAT]</span></td></tr> | ||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[ | |||
</table> | </table> | ||
== Function == | == Function == | ||
[ | [https://www.uniprot.org/uniprot/CLCD_PSEKB CLCD_PSEKB] Ring cleavage of cyclic ester dienelactone to produce maleylacetate. | ||
<div style="background-color:#fffaf0;"> | <div style="background-color:#fffaf0;"> | ||
== Publication Abstract from PubMed == | == Publication Abstract from PubMed == | ||
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__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: | [[Category: Pseudomonas knackmussii]] | ||
[[Category: | [[Category: Collyer CA]] | ||
[[Category: | [[Category: Ollis DL]] | ||
[[Category: | [[Category: Porter JL]] | ||
Latest revision as of 03:49, 28 December 2023
Crystal structure of dienelactone hydrolase B-1 variant (Q35H, F38L, Y64H, Q110L, C123S, Y137C, Y145C, N154D, E199G, S208G and G211D) at 1.80 A resolutionCrystal structure of dienelactone hydrolase B-1 variant (Q35H, F38L, Y64H, Q110L, C123S, Y137C, Y145C, N154D, E199G, S208G and G211D) at 1.80 A resolution
Structural highlights
FunctionCLCD_PSEKB Ring cleavage of cyclic ester dienelactone to produce maleylacetate. Publication Abstract from PubMedThe ease with which enzymes can be adapted from their native roles and engineered to function specifically for industrial or commercial applications is crucial to enabling enzyme technology to advance beyond its current state. Directed evolution is a powerful tool for engineering enzymes with improved physical and catalytic properties and can be used to evolve enzymes where lack of structural information may thwart the use of rational design. In this study, we take the versatile and diverse alpha/beta hydrolase fold framework, in the form of dienelactone hydrolase, and evolve it over three unique sequential evolutions with a total of 14 rounds of screening to generate a series of enzyme variants. The native enzyme has a low level of promiscuous activity toward p-nitrophenyl acetate but almost undetectable activity toward larger p-nitrophenyl esters. Using p-nitrophenyl acetate as an evolutionary intermediate, we have generated variants with altered specificity and catalytic activity up to 3 orders of magnitude higher than the native enzyme toward the larger nonphysiological p-nitrophenyl ester substrates. Several variants also possess increased stability resulting from the multidimensional approach to screening. Crystal structure analysis and substrate docking show how the enzyme active site changes over the course of the evolutions as either a direct or an indirect result of mutations. Directed Evolution of New and Improved Enzyme Functions Using an Evolutionary Intermediate and Multidirectional Search.,Porter JL, Boon PL, Murray TP, Huber T, Collyer CA, Ollis DL ACS Chem Biol. 2014 Dec 5. PMID:25419863[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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