View source for 9qez


==Carbonic anhydrase mutant==
<StructureSection load='9qez' size='340' side='right'caption='[[9qez]], [[Resolution|resolution]] 2.10&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[9qez]] is a 10 chain structure with sequence from [https://en.wikipedia.org/wiki/Caloramator_australicus_RC3 Caloramator australicus RC3] and [https://en.wikipedia.org/wiki/Unidentified Unidentified]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=9QEZ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=9QEZ 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]] 2.1&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=EPE:4-(2-HYDROXYETHYL)-1-PIPERAZINE+ETHANESULFONIC+ACID'>EPE</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</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=9qez FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=9qez OCA], [https://pdbe.org/9qez PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=9qez RCSB], [https://www.ebi.ac.uk/pdbsum/9qez PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=9qez ProSAT]</span></td></tr>
</table>
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
The 16S microbial community profiling of a metagenomics library from geothermal spring at Lisvori (Lesvos island, Greece) enabled the identification of a putative sequence exhibiting 95% identity to the gamma-type carbonic anhydrase (gamma-CA) from Caloramator australicus (gamma-CaCA). The sequence of gamma-CaCA was amplified by PCR, cloned, and expressed in E. coli. Activity assays showed that gamma-CaCA possesses very low, but detectable, anhydrase activity, while exhibiting no measurable esterase activity. Differential scanning fluorimetry (DSF) revealed that the enzyme shows high thermal stability with a melting temperature (T(m)) approximately 65-75 degrees C in the pH range between 5.5 and 9.0. The structure of gamma-CaCA was determined by X-ray crystallography at 1.11 A resolution, the highest resolution reported so far for a gamma-CA. The enzyme was crystallized as a trimer in the crystallographic asymmetric unit and contains three zinc-binding sites, one at each interface of neighboring subunits of the trimer. Structure-based rational design enabled the design and creation of a mutant enzyme (gamma-CaCAmut) which possessed a heptapeptide insertion at the active-site loop and two-point mutations. Kinetic analysis demonstrated that gamma-CaCAmut was successfully converted into a catalytically active esterase indicating successful activity gain through structure-guided engineering. The thermostability of gamma-CaCAmut was significantly increased, aligning with the thermostability typically observed in hyperthermostable enzymes. X-ray crystallographic analysis of the gamma-CaCAmut structure at 2.1 A resolution, provided detailed structural insights into how the mutations impact the overall enzyme structure, function, and thermostability. These findings provide valuable structural and functional insights into gamma-CAs and demonstrate a strategy for converting an inactive enzyme into a catalytically active form through rational design.

Structural and functional characterization of a metagenomically derived gamma-type carbonic anhydrase and its engineering into a hyperthermostable esterase.,Bodourian CS, Imran M, Georgakis ND, Papageorgiou AC, Labrou NE Protein Sci. 2025 Dec;34(12):e70396. doi: 10.1002/pro.70396. PMID:41294346<ref>PMID:41294346</ref>

From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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<div class="pdbe-citations 9qez" style="background-color:#fffaf0;"></div>
== References ==
<references/>
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</StructureSection>
[[Category: Caloramator australicus RC3]]
[[Category: Large Structures]]
[[Category: Unidentified]]
[[Category: Mohsin I]]
[[Category: Papageorgiou AC]]