View source for 9vig


==Crystal structure of fused glycerol dehydratase L113W variant==
<StructureSection load='9vig' size='340' side='right'caption='[[9vig]], [[Resolution|resolution]] 3.02&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[9vig]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Klebsiella_pneumoniae Klebsiella pneumoniae]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=9VIG OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=9VIG 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]] 3.02&#8491;</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=9vig FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=9vig OCA], [https://pdbe.org/9vig PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=9vig RCSB], [https://www.ebi.ac.uk/pdbsum/9vig PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=9vig ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/Q59476_KLEPN Q59476_KLEPN] [https://www.uniprot.org/uniprot/O08505_KLEPN O08505_KLEPN] 
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
Glycerol dehydratase (GDHt) enables bioconversion of glycerol to valuable chemicals, but its industrial use is hindered by rapid loss of the adenosylcobalamin (AdoCbl) cofactor (coenzyme B(12)) through both oxygen- and mechanism-based inactivation. To overcome this limitation, we reinforced the AdoCbl-binding interface of Klebsiella pneumoniae GDHt by fusing its alpha and beta subunits with a five-residue linker (fGDHt) and then introducing interface mutations. Fusion alone doubled the oxygen-inactivation half-life without affecting catalytic efficiency. Structural and computational analyses of interface residues, followed by experimental screening, yielded three stabilizing substitutions-alpha-A177M, beta-L113W, and beta-M158W. Pairwise combinations of these mutations yielded double variants whose oxygen-inactivation half-lives increased by up to 24-fold. Enzyme-coupled reactions to convert glycerol into 3-hydroxypropionic acid (3-HP) confirmed that engineered fGDHt variants maintained catalytic activity for longer periods, implying protection against both inactivation modes. In recombinant Escherichia coli strains producing 3-HP, the alpha-A177M/beta-M158W variant matched wild-type titers while operating with 25-fold less AdoCbl. Crystal structures reveal that the mutations tighten inter-subunit packing and, in the case of alpha-A177M, partly occlude an O(2)-access tunnel to the cofactor. These results have established alpha-beta interface engineering as a strategy for engineering more robust GDHts.

Engineering the alpha- and beta-subunit interface of a coenzyme B(12)-dependent glycerol dehydratase for enhancing its resistance to inactivation.,Na CY, Nasir A, Park R, Yeon YJ, Baek SH, Park S, Park YS, Seo MD, Yoo TH Bioresour Technol. 2026 Feb;442:133733. doi: 10.1016/j.biortech.2025.133733. Epub , 2025 Nov 28. PMID:41319883<ref>PMID:41319883</ref>

From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
<div class="pdbe-citations 9vig" style="background-color:#fffaf0;"></div>
== References ==
<references/>
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</StructureSection>
[[Category: Klebsiella pneumoniae]]
[[Category: Large Structures]]
[[Category: Park RY]]
[[Category: Seo MD]]