View source for 7p76


==Re-engineered 2-deoxy-D-ribose-5-phosphate aldolase catalysing asymmetric Michael addition reactions, Schiff base complex with cinnamaldehyde==
<StructureSection load='7p76' size='340' side='right'caption='[[7p76]], [[Resolution|resolution]] 1.90&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[7p76]] is a 12 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli_909945-2 Escherichia coli 909945-2]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7P76 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7P76 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]] 1.9&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=9Y6:(2E)-3-phenylprop-2-enal'>9Y6</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</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=7p76 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7p76 OCA], [https://pdbe.org/7p76 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7p76 RCSB], [https://www.ebi.ac.uk/pdbsum/7p76 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7p76 ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/V0AAC4_ECOLX V0AAC4_ECOLX] Catalyzes a reversible aldol reaction between acetaldehyde and D-glyceraldehyde 3-phosphate to generate 2-deoxy-D-ribose 5-phosphate.[HAMAP-Rule:MF_00592]
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== Publication Abstract from PubMed ==
Class I aldolases catalyze asymmetric aldol addition reactions and have found extensive application in the biocatalytic synthesis of chiral beta-hydroxy-carbonyl compounds. However, the usefulness of these powerful enzymes for application in other C-C bond-forming reactions remains thus far unexplored. The redesign of class I aldolases to expand their catalytic repertoire to include non-native carboligation reactions therefore continues to be a major challenge. Here, we report the successful redesign of 2-deoxy-d-ribose-5-phosphate aldolase (DERA) from Escherichia coli, an archetypical class I aldolase, to proficiently catalyze enantioselective Michael additions of nitromethane to alpha,beta-unsaturated aldehydes to yield various pharmaceutically relevant chiral synthons. After 11 rounds of directed evolution, the redesigned DERA enzyme (DERA-MA) carried 12 amino-acid substitutions and had an impressive 190-fold enhancement in catalytic activity compared to the wildtype enzyme. The high catalytic efficiency of DERA-MA for this abiological reaction makes it a proficient "Michaelase" with potential for biocatalytic application. Crystallographic analysis provides a structural context for the evolved activity. Whereas an aldolase acts naturally by activating the enzyme-bound substrate as a nucleophile (enamine-based mechanism), DERA-MA instead acts by activating the enzyme-bound substrate as an electrophile (iminium-based mechanism). This work demonstrates the power of directed evolution to expand the reaction scope of natural aldolases to include asymmetric Michael addition reactions and presents opportunities to explore iminium catalysis with DERA-derived catalysts inspired by developments in the organocatalysis field.

Unlocking Asymmetric Michael Additions in an Archetypical Class I Aldolase by Directed Evolution.,Kunzendorf A, Xu G, van der Velde JJH, Rozeboom HJ, Thunnissen AWH, Poelarends GJ ACS Catal. 2021 Nov 5;11(21):13236-13243. doi: 10.1021/acscatal.1c03911. Epub, 2021 Oct 15. PMID:34765282<ref>PMID:34765282</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 7p76" style="background-color:#fffaf0;"></div>

==See Also==
*[[Aldolase 3D structures|Aldolase 3D structures]]
== References ==
<references/>
__TOC__
</StructureSection>
[[Category: Escherichia coli 909945-2]]
[[Category: Large Structures]]
[[Category: Kunzendorf A]]
[[Category: Poelarends GJ]]
[[Category: Rozeboom HJ]]
[[Category: Thunnissen AMWH]]