Cloned (Comment) | Organism |
---|---|
the fragment is expressed in both BL21 (DE3) and B834 (DE3) Escherichia coli cells | Streptococcus pneumoniae |
Crystallization (Comment) | Organism |
---|---|
hangingdrop and sitting-drop vapour-diffusion methods | Streptococcus pneumoniae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
30000 | - |
- |
Streptococcus pneumoniae |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus pneumoniae | - |
- |
- |
Purification (Comment) | Organism |
---|---|
the cell pellet is resuspended in 50 ml lysis buffer (20 mM Tris-HCl pH 8.0 and 300 mM sodium chloride) and lysed by sonication on ice. All subsequent purification steps are performed at 277 K. The lysate is centrifuged at 15 000g for 30 min. The supernatant is applied onto a 10 ml Ni2+-NTA affinity column (Qiagen) equilibrated with lysis buffer. Nonspecifically bound proteins are washed from the column using 200 ml lysis buffer containing 15 mM imidazole. The recombinant protein is then eluted from the column with 20 ml elution buffer containing 20 mM Tris-HCl pH 8.0, 300 mM sodium chloride and 400 mM imidazole. The protein is concentrated and buffer-exchanged to the final buffer (5 mM Tris-HCl pH 8.0 and 10 mM sodium chloride). The final purified protein concentration was about 50 mg/ml and the purity is determined by SDS-PAGE to be about 95% | Streptococcus pneumoniae |
Synonyms | Comment | Organism |
---|---|---|
lysozyme SP0987 | - |
Streptococcus pneumoniae |