Refolding Record:
| Protein | |
|---|---|
| Protein Name | Beta-lactamase inhibitory protein II |
| Abbreviated Name | BLIP-II |
| SCOP Family | Beta-lactamase inhibitor protein-II, BLIP-II |
| Structure Notes | |
| Organism | Streptomyces exfoliatus |
| UniProt Accession | O87916 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 316 |
| Molecular Weight | 31056.5 |
| Pi | 4.7646 |
| Molecular Weight | 31056.5 |
| Disulphides | 0 |
| Full Sequence |
MPNGIVRLHRFGAALVMSGLLSALSAAPGVAAEPSSVSVAATSVVAWGGNNDWGEATVPA
EAQSGVDAIAGGYFHGLALKGGKVLGWGANLNGQLTMPAATQSGVDAIAAGNYHSLALKD
GEVIAWGGNEDGQTTVPAEARSGVDAIAAGAWASYALKDGKVIAWGDDSDGQTTVPAEAQ
SGVTALDGGVYTALAVKNGGVIAWGDNYFGQTTVPAEAQSGVDDVAGGIFHSLALKDGKV
IAWGDNRYKQTTVPTEALSGVSAIASGEWYSLALKNGKVIAWGSSRTAPSSVQSGVSSIE
AGPNAAYALKG
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Park HU, Lee KJ. (1998) Microbiology, 144, 2161-2167 |
| Project Aim | Undefined |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21 |
| Expression Temp | 37.0 |
| Expression Time | 3h |
| Expression Vector | pET3a |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | 50mM Tris-HCl, 10mM EDTA, 0.5% Triton X-100, pH 8.0 |
| Solubilization Buffer | 50mM Tris-HCl, 1mM EDTA, 100mM NaCl, 4M urea, pH 8.0 |
| Refolding Buffer | 5mM potassium phosphate |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The solubilized inclusion bodies were refolded by stepwise dilution. The concentration of urea was reduced two-fold at each step and the solution was then concentrated prior to the next dilution. The process was repeated 5 times in total and the protein was finally dialysed against 5mM phosphate buffer pH 7.0. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | |
| Notes | |