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| | ==Crystal structure of substrate-free CYP109A2 from Bacillus megaterium== | | ==Crystal structure of substrate-free CYP109A2 from Bacillus megaterium== |
| - | <StructureSection load='5ofq' size='340' side='right' caption='[[5ofq]], [[Resolution|resolution]] 2.70Å' scene=''> | + | <StructureSection load='5ofq' size='340' side='right'caption='[[5ofq]], [[Resolution|resolution]] 2.70Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5ofq]] is a 4 chain structure with sequence from [http://en.wikipedia.org/wiki/Bacmd Bacmd]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5OFQ OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5OFQ FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5ofq]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Priestia_megaterium_DSM_319 Priestia megaterium DSM 319]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5OFQ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5OFQ FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=1PE:PENTAETHYLENE+GLYCOL'>1PE</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.7Å</td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">BMD_2035 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=592022 BACMD])</td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=1PE:PENTAETHYLENE+GLYCOL'>1PE</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></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=5ofq FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5ofq OCA], [http://pdbe.org/5ofq PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5ofq RCSB], [http://www.ebi.ac.uk/pdbsum/5ofq PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5ofq ProSAT]</span></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=5ofq FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5ofq OCA], [https://pdbe.org/5ofq PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5ofq RCSB], [https://www.ebi.ac.uk/pdbsum/5ofq PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5ofq ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/D5DF88_PRIM3 D5DF88_PRIM3] |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | </div> | | </div> |
| | <div class="pdbe-citations 5ofq" style="background-color:#fffaf0;"></div> | | <div class="pdbe-citations 5ofq" style="background-color:#fffaf0;"></div> |
| | + | |
| | + | ==See Also== |
| | + | *[[Cytochrome P450 3D structures|Cytochrome P450 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Bacmd]] | + | [[Category: Large Structures]] |
| - | [[Category: Jozwik, I K]] | + | [[Category: Priestia megaterium DSM 319]] |
| - | [[Category: Thunnissen, A M.W H]]
| + | [[Category: Jozwik IK]] |
| - | [[Category: 25-hydroxyvitamin d3]]
| + | [[Category: Thunnissen AMWH]] |
| - | [[Category: Bacillus]]
| + | |
| - | [[Category: Bacillus megaterium]]
| + | |
| - | [[Category: Bacterial protein]] | + | |
| - | [[Category: Biocatalysis]] | + | |
| - | [[Category: Calcifediol]]
| + | |
| - | [[Category: Cholecalciferol]]
| + | |
| - | [[Category: Cytochrome p-450 enzyme system]]
| + | |
| - | [[Category: Cytochrome p450]]
| + | |
| - | [[Category: Heme]]
| + | |
| - | [[Category: Hydroxylation]]
| + | |
| - | [[Category: Oxidation-reduction]]
| + | |
| - | [[Category: Oxidoreductase]]
| + | |
| Structural highlights
Function
D5DF88_PRIM3
Publication Abstract from PubMed
Cytochrome P450 enzymes are increasingly investigated due to their potential application as biocatalysts with high regio- and/or stereo-selectivity and under mild conditions. Vitamin D3 metabolites are of pharmaceutical importance and are applied for the treatment of vitamin D3 deficiency and other disorders. However, the chemical synthesis of vitamin D3 derivatives shows low specificity and low yields. In this study, cytochrome P450 CYP109A2 from Bacillus megaterium DSM319 was expressed, purified and shown to oxidize vitamin D3 with high regio-selectivity. The in vitro conversion, using cytochrome P450 reductase (BmCPR) and ferredoxin (Fdx2) from the same strain, showed typical Michaelis-Menten reaction kinetics. A whole-cell system in B. megaterium overexpressing CYP109A2 reached 76 +/- 5% conversion after 24h and allowed to identify the main product by NMR analysis as 25-hydroxylated vitamin D3 . Product yield amounted to 54.9 mg L-1 day-1 , rendering the established whole-cell system as a highly promising biocatalytic route for the production of this valuable metabolite. The crystal structure of substrate-free CYP109A2 was determined at 2.7 A resolution, displaying an open conformation. Structural analysis predicts that CYP109A2 uses a highly similar set of residues for vitamin D3 binding as the related vitamin D3 hydroxylases CYP109E1 from Bacillus megaterium and CYP107BR1 (Vdh) from Pseudonocardia autotrophica. However, the folds and sequences of the BC loops in these three P450s are highly divergent, leading to differences in the shape and apolar/polar surface distribution of their active site pockets, which may account for the observed differences in substrate specificity and the regio-selectivity of vitamin D3 hydroxylation. This article is protected by copyright. All rights reserved.
Biochemical and structural characterization of CYP109A2, a vitamin D3 25-hydroxylase from Bacillus megaterium.,Abdulmughni A, Jozwik IK, Brill E, Hannemann F, Thunnissen AWH, Bernhardt R FEBS J. 2017 Sep 22. doi: 10.1111/febs.14276. PMID:28940959[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Abdulmughni A, Jozwik IK, Brill E, Hannemann F, Thunnissen AWH, Bernhardt R. Biochemical and structural characterization of CYP109A2, a vitamin D3 25-hydroxylase from Bacillus megaterium. FEBS J. 2017 Sep 22. doi: 10.1111/febs.14276. PMID:28940959 doi:http://dx.doi.org/10.1111/febs.14276
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