2e9b: Difference between revisions

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[[Image:2e9b.jpg|left|200px]]
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[[Image:2e9b.png|left|200px]]


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{{STRUCTURE_2e9b|  PDB=2e9b  |  SCENE=  }}  
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'''Crystal structure of pullulanase type I from Bacillus subtilis str. 168 complexed with maltose'''
===Crystal structure of pullulanase type I from Bacillus subtilis str. 168 complexed with maltose===


==Overview==
The AmyX gene encoding pullulanase from the common spore-forming bacterium Bacillus subtilis strain 168 was cloned, overexpressed in Escherichia coli, purified and crystallized. The recombinant pullulanase was purified to homogeneity using ammonium sulfate precipitation, hydrophobic chromatography and anion-exchange chromatography, resulting in a specific activity of 24.10 U per milligram of protein. SDS-PAGE analysis showed that the molecular weight of the protein is approximately 81.0 kDa, which is similar to the calculated molecular weight, 81.1 kDa, from its translated cDNA sequence. The k(cat) and K(m) of the purified enzyme with pullulan as substrate were approximately 79 s(-1) and 1.284 mg ml(-1), respectively. X-ray crystallographic analysis of the pullulanase crystal showed that the crystal belongs to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 70.568, b = 127.68, c = 189.25 angstroms. The crystal contains two molecules of pullulanase in the asymmetric unit, with a solvent content of 53.15%. The crystal diffracted to 2.1 angstroms resolution at a synchrotron and is suitable for structure determination.


==About this Structure==
==About this Structure==
2E9B is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2E9B OCA].  
2E9B is a 2 chains structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2E9B OCA].  


==Reference==
==Reference==
Overexpression, purification and preliminary X-ray analysis of pullulanase from Bacillus subtilis strain 168., Malle D, Itoh T, Hashimoto W, Murata K, Utsumi S, Mikami B, Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Apr 1;62(Pt, 4):381-4. Epub 2006 Mar 25. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16582490 16582490]
<ref group="xtra">PMID:16582490</ref><references group="xtra"/>
[[Category: Bacillus subtilis]]
[[Category: Bacillus subtilis]]
[[Category: Pullulanase]]
[[Category: Pullulanase]]
[[Category: Single protein]]
[[Category: Iwamoto, H.]]
[[Category: Iwamoto, H.]]
[[Category: Katsuya, Y.]]
[[Category: Katsuya, Y.]]
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[[Category: Hydrolase]]
[[Category: Hydrolase]]
[[Category: Smultiple domain]]
[[Category: Smultiple domain]]
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May  4 02:10:42 2008''
 
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Feb 16 20:08:43 2009''

Revision as of 18:08, 16 February 2009

File:2e9b.png

Template:STRUCTURE 2e9b

Crystal structure of pullulanase type I from Bacillus subtilis str. 168 complexed with maltose

About this Structure

2E9B is a 2 chains structure of sequences from Bacillus subtilis. Full crystallographic information is available from OCA.

Reference

  1. Malle D, Itoh T, Hashimoto W, Murata K, Utsumi S, Mikami B. Overexpression, purification and preliminary X-ray analysis of pullulanase from Bacillus subtilis strain 168. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Apr 1;62(Pt, 4):381-4. Epub 2006 Mar 25. PMID:16582490 doi:10.1107/S1744309106007901

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