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==Crystal structure of glyceraldehyde-3-phosphate dehydrogenase from Corynebacterium glutamicum ATCC13032 (L36S/T37K/F100V/P192S) in complex with NADP== | |||
<StructureSection load='8hrt' size='340' side='right'caption='[[8hrt]], [[Resolution|resolution]] 1.99Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[8hrt]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Corynebacterium_glutamicum Corynebacterium glutamicum]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=8HRT OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=8HRT 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]] 1.99Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=NAP:NADP+NICOTINAMIDE-ADENINE-DINUCLEOTIDE+PHOSPHATE'>NAP</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=8hrt FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=8hrt OCA], [https://pdbe.org/8hrt PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=8hrt RCSB], [https://www.ebi.ac.uk/pdbsum/8hrt PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=8hrt ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Since the discovery of l-glutamate-producing Corynebacterium glutamicum, it has evolved to be an industrial workhorse. For biobased chemical production, suppling sufficient amounts of the NADPH cofactor is crucial. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme that converts glyceraldehyde-3-phosphate (G3P) to 1,3-bisphosphoglycerate and produces NADH, is a major prospective solution for the cofactor imbalance issue. In this study, we determined the crystal structure of GAPDH from C. glutamicum ATCC13032 (CgGAPDH). Based on the structural information, we generated six CgGAPDH variants, CgGAPDH(L36S), CgGAPDH(L36S/T37K), CgGAPDH(L36S/T37K/P192S), CgGAPDH(L36S/T37K/F100V/P192S), CgGAPDH(L36S/T37K/F100L/P192S), and CgGAPDH(L36S/T37K/F100I/P192S), that can produce both NADH and NAPDH. The final CgGAPDH(L36S/T37K/F100V/P192S) variant showed a 212-fold increase in enzyme activity for NADP as well as 200% and 30% increased activity for the G3P substrate under NAD and NADP cofactor conditions, respectively. In addition, crystal structures of CgGAPDH variants in complex with NAD(P) permit the elucidation of differences between wild-type CgGAPDH and variants in relation to cofactor stabilization. | |||
Structure-Guided Protein Engineering of Glyceraldehyde-3-phosphate Dehydrogenase from Corynebacterium glutamicum for Dual NAD/NADP Cofactor Specificity.,Son HF, Yu H, Hong J, Lee D, Kim IK, Kim KJ J Agric Food Chem. 2023 Nov 22;71(46):17852-17859. doi: 10.1021/acs.jafc.3c06176. , Epub 2023 Nov 7. PMID:37935620<ref>PMID:37935620</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
<div class="pdbe-citations 8hrt" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Corynebacterium glutamicum]] | |||
[[Category: Large Structures]] | |||
[[Category: Kim KJ]] | |||
[[Category: Son HF]] | |||
Latest revision as of 11:01, 6 December 2023
Crystal structure of glyceraldehyde-3-phosphate dehydrogenase from Corynebacterium glutamicum ATCC13032 (L36S/T37K/F100V/P192S) in complex with NADPCrystal structure of glyceraldehyde-3-phosphate dehydrogenase from Corynebacterium glutamicum ATCC13032 (L36S/T37K/F100V/P192S) in complex with NADP
Structural highlights
Publication Abstract from PubMedSince the discovery of l-glutamate-producing Corynebacterium glutamicum, it has evolved to be an industrial workhorse. For biobased chemical production, suppling sufficient amounts of the NADPH cofactor is crucial. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme that converts glyceraldehyde-3-phosphate (G3P) to 1,3-bisphosphoglycerate and produces NADH, is a major prospective solution for the cofactor imbalance issue. In this study, we determined the crystal structure of GAPDH from C. glutamicum ATCC13032 (CgGAPDH). Based on the structural information, we generated six CgGAPDH variants, CgGAPDH(L36S), CgGAPDH(L36S/T37K), CgGAPDH(L36S/T37K/P192S), CgGAPDH(L36S/T37K/F100V/P192S), CgGAPDH(L36S/T37K/F100L/P192S), and CgGAPDH(L36S/T37K/F100I/P192S), that can produce both NADH and NAPDH. The final CgGAPDH(L36S/T37K/F100V/P192S) variant showed a 212-fold increase in enzyme activity for NADP as well as 200% and 30% increased activity for the G3P substrate under NAD and NADP cofactor conditions, respectively. In addition, crystal structures of CgGAPDH variants in complex with NAD(P) permit the elucidation of differences between wild-type CgGAPDH and variants in relation to cofactor stabilization. Structure-Guided Protein Engineering of Glyceraldehyde-3-phosphate Dehydrogenase from Corynebacterium glutamicum for Dual NAD/NADP Cofactor Specificity.,Son HF, Yu H, Hong J, Lee D, Kim IK, Kim KJ J Agric Food Chem. 2023 Nov 22;71(46):17852-17859. doi: 10.1021/acs.jafc.3c06176. , Epub 2023 Nov 7. PMID:37935620[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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