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4eez

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==Crystal Structure of Lactococcus lactis Alcohol Dehydrogenase variant RE1==
==Crystal Structure of Lactococcus lactis Alcohol Dehydrogenase variant RE1==
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<StructureSection load='4eez' size='340' side='right' caption='[[4eez]], [[Resolution|resolution]] 1.90&Aring;' scene=''>
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<StructureSection load='4eez' size='340' side='right'caption='[[4eez]], [[Resolution|resolution]] 1.90&Aring;' scene=''>
== Structural highlights ==
== Structural highlights ==
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<table><tr><td colspan='2'>[[4eez]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Lactococcus_lactis_subsp._lactis_kf147 Lactococcus lactis subsp. lactis kf147]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4EEZ OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4EEZ FirstGlance]. <br>
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<table><tr><td colspan='2'>[[4eez]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Lactococcus_lactis_subsp._lactis_KF147 Lactococcus lactis subsp. lactis KF147]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4EEZ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4EEZ FirstGlance]. <br>
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</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=PG4:TETRAETHYLENE+GLYCOL'>PG4</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>
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</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.9&#8491;</td></tr>
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<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4eex|4eex]]</td></tr>
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<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=PG4:TETRAETHYLENE+GLYCOL'>PG4</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>
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<tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">adhA, LLKF_1981 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=684738 Lactococcus lactis subsp. lactis KF147])</td></tr>
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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=4eez FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4eez OCA], [https://pdbe.org/4eez PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4eez RCSB], [https://www.ebi.ac.uk/pdbsum/4eez PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4eez ProSAT]</span></td></tr>
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<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Alcohol_dehydrogenase Alcohol dehydrogenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.1 1.1.1.1] </span></td></tr>
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<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=4eez FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4eez OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4eez RCSB], [http://www.ebi.ac.uk/pdbsum/4eez PDBsum]</span></td></tr>
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</table>
</table>
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<div style="background-color:#fffaf0;">
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== Function ==
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== Publication Abstract from PubMed ==
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[https://www.uniprot.org/uniprot/D2BLA0_LACLK D2BLA0_LACLK]
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We have determined the X-ray crystal structures of the NADH-dependent alcohol dehydrogenase LlAdhA from Lactococcus lactis and its laboratory-evolved variant LlAdhA(RE1) at 1.9A and 2.5A resolution, respectively. LlAdhA(RE1), which contains three amino acid mutations (Y50F, I212T, and L264V), was engineered to increase the microbial production of isobutanol (2-methylpropan-1-ol) from isobutyraldehyde (2-methylpropanal). Structural comparison of LlAdhA and LlAdhA(RE1) indicates that the enhanced activity on isobutyraldehyde stems from increases in the protein's active site size, hydrophobicity, and substrate access. Further structure-guided mutagenesis generated a quadruple mutant (Y50F/N110S/I212T/L264V), whose K(M) for isobutyraldehyde is approximately 17-fold lower and catalytic efficiency (k(cat)/K(M)) is approximately 160-fold higher than wild-type LlAdhA. Combining detailed structural information and directed evolution, we have achieved significant improvements in non-native alcohol dehydrogenase activity that will facilitate the production of next-generation fuels such as isobutanol from renewable resources.
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Structure-guided engineering of Lactococcus lactis alcohol dehydrogenase LlAdhA for improved conversion of isobutyraldehyde to isobutanol.,Liu X, Bastian S, Snow CD, Brustad EM, Saleski TE, Xu JH, Meinhold P, Arnold FH J Biotechnol. 2012 Sep 3. pii: S0168-1656(12)00610-4. doi:, 10.1016/j.jbiotec.2012.08.008. PMID:22974724<ref>PMID:22974724</ref>
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From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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</div>
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==See Also==
==See Also==
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*[[Alcohol dehydrogenase|Alcohol dehydrogenase]]
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*[[Alcohol dehydrogenase 3D structures|Alcohol dehydrogenase 3D structures]]
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== References ==
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<references/>
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__TOC__
__TOC__
</StructureSection>
</StructureSection>
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[[Category: Alcohol dehydrogenase]]
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[[Category: Lactococcus lactis subsp. lactis KF147]]
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[[Category: Lactococcus lactis subsp. lactis kf147]]
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[[Category: Large Structures]]
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[[Category: Arnold, F H]]
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[[Category: Arnold FH]]
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[[Category: Bastian, S]]
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[[Category: Bastian S]]
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[[Category: Brustad, E M]]
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[[Category: Brustad EM]]
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[[Category: Liu, X]]
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[[Category: Liu X]]
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[[Category: Meinhold, P]]
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[[Category: Meinhold P]]
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[[Category: Saleski, T]]
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[[Category: Saleski T]]
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[[Category: Snow, C D]]
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[[Category: Snow CD]]
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[[Category: Xu, J H]]
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[[Category: Xu JH]]
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[[Category: Biofuel]]
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[[Category: Directed evolution]]
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[[Category: Isobutyraldehyde]]
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[[Category: Oxidoreductase]]
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[[Category: Site-saturation mutagenesis]]
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Current revision

Crystal Structure of Lactococcus lactis Alcohol Dehydrogenase variant RE1

PDB ID 4eez

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