2e9b
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(New page: 200px<br /><applet load="2e9b" size="350" color="white" frame="true" align="right" spinBox="true" caption="2e9b, resolution 2.30Å" /> '''Crystal structure of...)
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Revision as of 15:07, 21 February 2008
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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
2E9B is a Single protein structure of sequence from Bacillus subtilis with , and as ligands. Active as Pullulanase, with EC number 3.2.1.41 Known structural/functional Sites: , , , , , , , , , , , , and . Full crystallographic information is available from OCA.
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:16582490
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