Expression, crystallization and preliminary X-ray crystallographic studies of<i>Klebsiella pneumoniae</i>maltohexaose-producing α-amylase
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説明
A recombinant form of Klebsiella pneumoniae maltohexaose-producing alpha-amylase has been overexpressed in Escherichia coli and purified to homogeneity. Crystals were obtained at 293 K by the microbatch technique using 80 mM sodium/potassium phosphate buffer pH 6.2 containing 8% polyethylene glycol 3000, 4% polyethylene glycol 3350 and 40 mM sodium thiocyanate. Crystals of the overexpressed recombinant enzyme diffracted to better than 2.5 A resolution at 95 K using a synchrotron-radiation source. The crystals belong to the primitive monoclinic space group P2(1), with unit-cell parameters a = 74.8, b = 107.6, c = 82.2 A, beta = 96.2 degrees. Assuming the presence of two molecules per asymmetric unit, the V(M) value for the crystal was 2.3 A(3) Da(-1), indicating a solvent content of 47%.
収録刊行物
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- Acta Crystallographica Section D Biological Crystallography
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Acta Crystallographica Section D Biological Crystallography 60 2352-2354, 2004-11-26
International Union of Crystallography (IUCr)
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キーワード
- Potassium Compounds
- Protein Conformation
- Temperature
- Oligosaccharides
- Buffers
- Hydrogen-Ion Concentration
- Crystallography, X-Ray
- Recombinant Proteins
- Phosphates
- Polyethylene Glycols
- Klebsiella pneumoniae
- X-Ray Diffraction
- Mutation
- Electrophoresis, Polyacrylamide Gel
- alpha-Amylases
- Crystallization
- Synchrotrons
- Thiocyanates