5br4: Difference between revisions
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''' | ==E. coli lactaldehyde reductase (FucO) M185C mutant== | ||
<StructureSection load='5br4' size='340' side='right' caption='[[5br4]], [[Resolution|resolution]] 0.91Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[5br4]] is a 2 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5BR4 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5BR4 FirstGlance]. <br> | |||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=NAD:NICOTINAMIDE-ADENINE-DINUCLEOTIDE'>NAD</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr> | |||
<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Lactaldehyde_reductase Lactaldehyde reductase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.77 1.1.1.77] </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=5br4 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5br4 OCA], [http://pdbe.org/5br4 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5br4 RCSB], [http://www.ebi.ac.uk/pdbsum/5br4 PDBsum]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
NAD(P)H-dependent enzymes are ubiquitous in metabolism and cellular processes and are also of great interest for pharmaceutical and industrial applications. Here, we present a structure-guided enzyme engineering strategy for improving catalytic properties of NAD(P)H-dependent enzymes toward native or native-like reactions using mutations to the enzyme's adenine-binding pocket, distal to the site of catalysis. Screening single-site saturation mutagenesis libraries identified mutations that increased catalytic efficiency up to 10-fold in 7 out of 10 enzymes. The enzymes improved in this study represent three different cofactor-binding folds (Rossmann, DHQS-like, and FAD/NAD binding) and utilize both NADH and NADPH. Structural and biochemical analyses show that the improved activities are accompanied by minimal changes in other properties (cooperativity, thermostability, pH optimum, uncoupling), and initial tests on two enzymes (ScADH6 and EcFucO) show improved functionality in Escherichia coli. | |||
Mutations in adenine-binding pockets enhance catalytic properties of NAD(P)H-dependent enzymes.,Cahn JK, Baumschlager A, Brinkmann-Chen S, Arnold FH Protein Eng Des Sel. 2016 Jan;29(1):31-8. doi: 10.1093/protein/gzv057. Epub 2015 , Oct 27. PMID:26512129<ref>PMID:26512129</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 5br4" style="background-color:#fffaf0;"></div> | |||
[[Category: | == References == | ||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Lactaldehyde reductase]] | |||
[[Category: Arnold, F H]] | |||
[[Category: Brinkmann-Chen, S]] | [[Category: Brinkmann-Chen, S]] | ||
[[Category: Cahn, J | [[Category: Cahn, J K.B]] | ||
[[Category: | [[Category: Fuco]] | ||
[[Category: Mutant]] | |||
[[Category: Nadh]] | |||
[[Category: Oxidoreductase]] | |||
Revision as of 15:42, 23 December 2015
E. coli lactaldehyde reductase (FucO) M185C mutantE. coli lactaldehyde reductase (FucO) M185C mutant
Structural highlights
Publication Abstract from PubMedNAD(P)H-dependent enzymes are ubiquitous in metabolism and cellular processes and are also of great interest for pharmaceutical and industrial applications. Here, we present a structure-guided enzyme engineering strategy for improving catalytic properties of NAD(P)H-dependent enzymes toward native or native-like reactions using mutations to the enzyme's adenine-binding pocket, distal to the site of catalysis. Screening single-site saturation mutagenesis libraries identified mutations that increased catalytic efficiency up to 10-fold in 7 out of 10 enzymes. The enzymes improved in this study represent three different cofactor-binding folds (Rossmann, DHQS-like, and FAD/NAD binding) and utilize both NADH and NADPH. Structural and biochemical analyses show that the improved activities are accompanied by minimal changes in other properties (cooperativity, thermostability, pH optimum, uncoupling), and initial tests on two enzymes (ScADH6 and EcFucO) show improved functionality in Escherichia coli. Mutations in adenine-binding pockets enhance catalytic properties of NAD(P)H-dependent enzymes.,Cahn JK, Baumschlager A, Brinkmann-Chen S, Arnold FH Protein Eng Des Sel. 2016 Jan;29(1):31-8. doi: 10.1093/protein/gzv057. Epub 2015 , Oct 27. PMID:26512129[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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