Refolding Record:
| Protein | |
|---|---|
| Protein Name | Bowman-Birk Inhibitor |
| Abbreviated Name | BBI |
| SCOP Family | Bowman-Birk Inhibitor |
| Structure Notes | |
| Organism | Rice |
| UniProt Accession | Q0JR25 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Small Proteins |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 258 |
| Molecular Weight | 27789.7 |
| Pi | 5.37887 |
| Molecular Weight | 27789.7 |
| Disulphides | 7 |
| Full Sequence |
MSNTTMATSTILLFLLAGLAAAHGDGDTTIRLPSDGAKASRPRAAKPWDCCDNIEISRLM
IYPPLYRCNDEVKQCAAACKECVEAPGGDFNGGAFVCSDWFSTVDPGPKCTAALDGLSME
RPWKCCDNIKRLPTKPDPPQWRCNDELEPSQCTAACKSCREAPGPFPGKLICEDIYWGAD
PGPFCTPRPWGDCCDKAFCNKMNPPTCRCMDEVKECADACKDCQRVESSEPPRYVCKDRF
TGHPGPVCKPRAEN
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Li N, Qu LJ, Liu Y, Li Q, Gu H, Chen Z. (1999) Protein Expression and Purification, 15, 99-104 |
| Project Aim | Undefined |
| Fusion | N-terminal hexahistidine tag |
| 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 | 4h |
| Expression Vector | pET28a |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Freeze/Thaw+Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | partial |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | 2M urea,, 50mM Tris-HCl, pH 8.0 |
| Solubilization Buffer | 8 M urea, 50 mM Tris?HCl, pH 8.0,100 mM b-mercaptoethanol |
| Refolding Buffer | 100 mM Tris?HCl, 0.5 M L-arginine, 1 mM GSSG |
| Pre-Refolding Purification | None |
| Tag Cleaved | yes |
| Refolding pH | 8.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | up to 0.3mg/ml |
| Refolding Time | 12-48h |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Renaturation was initiated by rapidly diluting 1.25 ml of the denatured and reduced protein into 125 ml refolding buffer (100 mM Tris?HCl, pH 8.0, 0.5 M L-arginine, 1 mM GSSG) (100-fold dilution). The sample was incubated without mixing at room temperature for about 12 h before another 1.25 ml of denatured proteins was added. As many as four aliquots could be added together until the final protein concentration reached about 300 mg/ml. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | 57% |
| Notes | |