3vm5: Difference between revisions
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The | ==Recombinant medaka fish alpha-amylase expressed in yeast Pichia pastoris== | ||
<StructureSection load='3vm5' size='340' side='right'caption='[[3vm5]], [[Resolution|resolution]] 2.85Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[3vm5]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Oryzias_latipes Oryzias latipes]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3VM5 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3VM5 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]] 2.85Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</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=3vm5 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3vm5 OCA], [https://pdbe.org/3vm5 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3vm5 RCSB], [https://www.ebi.ac.uk/pdbsum/3vm5 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3vm5 ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/AMY_ORYLA AMY_ORYLA] Catalyzes the hydrolysis of alpha-1,4 glycosidic linkages in starch, glycogen and similar oligosaccharides.<ref>PMID:22613096</ref> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
The medaka fish alpha-amylase was expressed and purified. The expression systems were constructed using methylotrophic yeast Pichia pastoris, and the recombinant proteins were secreted into the culture medium. Purified recombinant alpha-amylase exhibited starch hydrolysis activity. The optimal pH, denaturation temperature, and K(M) and V(max) values were determined; chloride ions were essential for enzyme activity. The purified protein was also crystallized and examined by X-ray crystallography. The structure has the (alpha/beta)(8) barrel fold, as do other known alpha-amylases, and the overall structure is very similar to the structure of vertebrate (human and pig) alpha-amylases. A novel expression plasmid was developed. Using this plasmid, high-throughput construction of an expression system by homologous recombination in P. pastoris cells, previously reported for membrane proteins, was successfully applied to the secretory protein. | |||
Structural and functional characterization of recombinant medaka fish alpha-amylase expressed in yeast Pichia pastoris.,Mizutani K, Toyoda M, Otake Y, Yoshioka S, Takahashi N, Mikami B Biochim Biophys Acta. 2012 May 18. PMID:22613096<ref>PMID:22613096</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 3vm5" style="background-color:#fffaf0;"></div> | |||
==See Also== | |||
*[[Amylase 3D structures|Amylase 3D structures]] | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Oryzias latipes]] | |||
[[Category: Mikami B]] | |||
[[Category: Mizutani K]] | |||
[[Category: Toyoda M]] | |||
Latest revision as of 15:26, 8 November 2023
Recombinant medaka fish alpha-amylase expressed in yeast Pichia pastorisRecombinant medaka fish alpha-amylase expressed in yeast Pichia pastoris
Structural highlights
FunctionAMY_ORYLA Catalyzes the hydrolysis of alpha-1,4 glycosidic linkages in starch, glycogen and similar oligosaccharides.[1] Publication Abstract from PubMedThe medaka fish alpha-amylase was expressed and purified. The expression systems were constructed using methylotrophic yeast Pichia pastoris, and the recombinant proteins were secreted into the culture medium. Purified recombinant alpha-amylase exhibited starch hydrolysis activity. The optimal pH, denaturation temperature, and K(M) and V(max) values were determined; chloride ions were essential for enzyme activity. The purified protein was also crystallized and examined by X-ray crystallography. The structure has the (alpha/beta)(8) barrel fold, as do other known alpha-amylases, and the overall structure is very similar to the structure of vertebrate (human and pig) alpha-amylases. A novel expression plasmid was developed. Using this plasmid, high-throughput construction of an expression system by homologous recombination in P. pastoris cells, previously reported for membrane proteins, was successfully applied to the secretory protein. Structural and functional characterization of recombinant medaka fish alpha-amylase expressed in yeast Pichia pastoris.,Mizutani K, Toyoda M, Otake Y, Yoshioka S, Takahashi N, Mikami B Biochim Biophys Acta. 2012 May 18. PMID:22613096[2] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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