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| | ==Crystal Structure of Lactococcus lactis Alcohol Dehydrogenase variant RE1== | | ==Crystal Structure of Lactococcus lactis Alcohol Dehydrogenase variant RE1== |
| - | <StructureSection load='4eez' size='340' side='right' caption='[[4eez]], [[Resolution|resolution]] 1.90Å' scene=''> | + | <StructureSection load='4eez' size='340' side='right'caption='[[4eez]], [[Resolution|resolution]] 1.90Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4eez]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Laclk Laclk]. 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> | + | <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> |
| - | </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> | + | </td></tr><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> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4eex|4eex]]</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=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> |
| - | <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 LACLK])</td></tr>
| + | |
| - | <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>
| + | |
| - | <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://pdbe.org/4eez PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4eez RCSB], [http://www.ebi.ac.uk/pdbsum/4eez PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4eez ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | + | == Function == |
| | + | [[https://www.uniprot.org/uniprot/D2BLA0_LACLK D2BLA0_LACLK]] |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | ==See Also== | | ==See Also== |
| - | *[[Alcohol dehydrogenase|Alcohol dehydrogenase]] | + | *[[Alcohol dehydrogenase 3D structures|Alcohol dehydrogenase 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Alcohol dehydrogenase]] | + | [[Category: Lactococcus lactis subsp. lactis KF147]] |
| - | [[Category: Laclk]] | + | [[Category: Large Structures]] |
| - | [[Category: Arnold, F H]] | + | [[Category: Arnold FH]] |
| - | [[Category: Bastian, S]] | + | [[Category: Bastian S]] |
| - | [[Category: Brustad, E M]] | + | [[Category: Brustad EM]] |
| - | [[Category: Liu, X]] | + | [[Category: Liu X]] |
| - | [[Category: Meinhold, P]] | + | [[Category: Meinhold P]] |
| - | [[Category: Saleski, T]] | + | [[Category: Saleski T]] |
| - | [[Category: Snow, C D]] | + | [[Category: Snow CD]] |
| - | [[Category: Xu, J H]] | + | [[Category: Xu JH]] |
| - | [[Category: Biofuel]]
| + | |
| - | [[Category: Directed evolution]]
| + | |
| - | [[Category: Isobutyraldehyde]]
| + | |
| - | [[Category: Oxidoreductase]]
| + | |
| - | [[Category: Site-saturation mutagenesis]]
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| Structural highlights
Function
[D2BLA0_LACLK]
Publication Abstract from PubMed
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.
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[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Liu X, Bastian S, Snow CD, Brustad EM, Saleski TE, Xu JH, Meinhold P, Arnold FH. Structure-guided engineering of Lactococcus lactis alcohol dehydrogenase LlAdhA for improved conversion of isobutyraldehyde to isobutanol. J Biotechnol. 2012 Sep 3. pii: S0168-1656(12)00610-4. doi:, 10.1016/j.jbiotec.2012.08.008. PMID:22974724 doi:http://dx.doi.org/10.1016/j.jbiotec.2012.08.008
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