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| | ==Crystal structure of CALB mutant D223G from Candida antarctica== | | ==Crystal structure of CALB mutant D223G from Candida antarctica== |
| - | <StructureSection load='4k6k' size='340' side='right' caption='[[4k6k]], [[Resolution|resolution]] 1.60Å' scene=''> | + | <StructureSection load='4k6k' size='340' side='right'caption='[[4k6k]], [[Resolution|resolution]] 1.60Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4k6k]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Canar Canar]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4K6K OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4K6K FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4k6k]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Moesziomyces_antarcticus Moesziomyces antarcticus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4K6K OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4K6K FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</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]] 1.6Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4k5q|4k5q]], [[4k6g|4k6g]], [[4k6h|4k6h]]</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></td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Triacylglycerol_lipase Triacylglycerol lipase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.1.3 3.1.1.3] </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=4k6k FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4k6k OCA], [https://pdbe.org/4k6k PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4k6k RCSB], [https://www.ebi.ac.uk/pdbsum/4k6k PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4k6k ProSAT]</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=4k6k FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4k6k OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4k6k RCSB], [http://www.ebi.ac.uk/pdbsum/4k6k PDBsum]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/LIPB_CANAR LIPB_CANAR]] Hydrolysis of triglycerides. Is very stereospecific both in hydrolysis and in organic synthesis and has a potentially important application in glucolipid synthesis. | + | [https://www.uniprot.org/uniprot/LIPB_PSEA2 LIPB_PSEA2] Hydrolysis of triglycerides. Is very stereospecific both in hydrolysis and in organic synthesis and has a potentially important application in glucolipid synthesis. |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| | </div> | | </div> |
| - | | + | <div class="pdbe-citations 4k6k" style="background-color:#fffaf0;"></div> |
| - | ==See Also== | + | |
| - | *[[Lipase|Lipase]]
| + | |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Canar]] | + | [[Category: Large Structures]] |
| - | [[Category: Triacylglycerol lipase]] | + | [[Category: Moesziomyces antarcticus]] |
| - | [[Category: An, J]] | + | [[Category: An J]] |
| - | [[Category: Feng, Y]] | + | [[Category: Feng Y]] |
| - | [[Category: Wu, G]] | + | [[Category: Wu G]] |
| - | [[Category: Xie, Y]] | + | [[Category: Xie Y]] |
| - | [[Category: Hydrolase]]
| + | |
| - | [[Category: Lipase]]
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| Structural highlights
Function
LIPB_PSEA2 Hydrolysis of triglycerides. Is very stereospecific both in hydrolysis and in organic synthesis and has a potentially important application in glucolipid synthesis.
Publication Abstract from PubMed
Enzyme stability is an important issue for protein engineers. Understanding how rigidity in the active site affects protein kinetic stability will provide new insight into enzyme stabilization. In this study, we demonstrated enhanced kinetic stability of Candida antarctica lipase B (CalB) by mutating the structurally flexible residues within the active site. Six residues within 10 A of the catalytic Ser(105) residue with a high B factor were selected for iterative saturation mutagenesis. After screening 2200 colonies, we obtained the D223G/L278M mutant, which exhibited a 13-fold increase in half-life at 48 degrees C and a 12 degrees C higher T50(15), the temperature at which enzyme activity is reduced to 50% after a 15-min heat treatment. Further characterization showed that global unfolding resistance against both thermal and chemical denaturation also improved. Analysis of the crystal structures of wild-type CalB and the D223G/L278M mutant revealed that the latter formed an extra main chain hydrogen bond network with seven structurally coupled residues within the flexible alpha10 helix that are primarily involved in forming the active site. Further investigation of the relative B factor profile and molecular dynamics simulation confirmed that the enhanced rigidity decreased fluctuation of the active site residues at high temperature. These results indicate that enhancing the rigidity of the flexible segment within the active site may provide an efficient method for improving enzyme kinetic stability.
Enhanced enzyme kinetic stability by increasing rigidity within the active site.,Xie Y, An J, Yang G, Wu G, Zhang Y, Cui L, Feng Y J Biol Chem. 2014 Mar 14;289(11):7994-8006. doi: 10.1074/jbc.M113.536045. Epub, 2014 Jan 21. PMID:24448805[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Xie Y, An J, Yang G, Wu G, Zhang Y, Cui L, Feng Y. Enhanced enzyme kinetic stability by increasing rigidity within the active site. J Biol Chem. 2014 Mar 14;289(11):7994-8006. doi: 10.1074/jbc.M113.536045. Epub, 2014 Jan 21. PMID:24448805 doi:http://dx.doi.org/10.1074/jbc.M113.536045
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