Refolding Record:
| Protein | |
|---|---|
| Protein Name | Beta-casein |
| Abbreviated Name | CA |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Bovine |
| UniProt Accession | P02666 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 210 |
| Molecular Weight | 23583.3 |
| Pi | 5.12 |
| Molecular Weight | 23583.3 |
| Disulphides | Unknown |
| Full Sequence |
RELEELNVPGEIVESLSSSEESITRINKKIEKFQSEEQQQTEDELQDKIHPFAQTQSLVYPFPGPIPNSLPQNIPPLTQTPVVVPPFLQPEVMGVSKVKEAMAPKHKEMPFPKYPVEPFTESQSLTLTDVENLHLPLPLLQSWMHQPHQPLPPTVMFPPQSVLSLSQSKVLPVPQKAVPYPQRDMPIQAFLLYQEPVLGPVRGPFPIIV
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Khodarahmi R, Beyrami M, Soori H. (2008) Archives Biochemistry and Biophysics, 1, 1 |
| Project Aim | Protein refolding |
| Fusion | None |
| Protein Expression and Production | Protein expressed and purified in native conformation prior to denaturation and refolding. |
| Expression Host | None |
| Expression Strain | None |
| Expression Temp | 0.0 |
| Expression Time | 0 |
| Expression Vector | |
| Expression Protocol | |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | 20 mM Tris–sulfate, pH 7.75, containing 6 M GuHCl |
| Refolding Buffer | 20 mM Tris–sulfate, pH 7.75 |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.75 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | 2-24 h |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Native CA (10 mg/ml) was denatured by overnight incubation in denaturation buffer (20 mM Tris–sulfate, pH 7.75, containing 6 M GuHCl) at room temperature. The denatured protein was stored at 4 °C and used within 2 weeks. Protein renaturation was conducted by rapid dilution of the denatured CA sample by 50-fold, using freshly prepared renaturation buffer. The composition of renaturation buffer with or without additive (casein) was similar to the denaturation buffer except for denaturant. The resulting renaturation systems were incubated at 25 °C for 2 h (or 24 h) followed by enzyme assay. |
| Refolding Assay | enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |