Refolding Record:
| Protein | |
|---|---|
| Protein Name | Lysozyme |
| Abbreviated Name | Lysozyme |
| SCOP Family | C-type Lysozyme |
| Structure Notes | |
| Organism | Chicken (Gallus gallus) |
| UniProt Accession | Q6LEL2 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha+Beta |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 45 |
| Molecular Weight | 4953.0 |
| Pi | 9.69 |
| Molecular Weight | 4953.0 |
| Disulphides | Unknown |
| Full Sequence |
MRSLLILVLCFLPLAALGKVFGRCELAAAMKRHGLDNYRGYSLGN
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Lange C, Patil G, Rudolph R. (2005) Protein Science, 14, 2693-701 |
| 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 | not specified |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | not indicated in paper |
| Refolding Buffer | (3 mM GSH, 0.3 mM GSSG, 1 mM EDTA, 0.1 M Tris/HCl [pH 8.2]), containing organic salts as indicated |
| Pre-Refolding Purification | not specified |
| Tag Cleaved | no |
| Refolding pH | 8.2 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | overnight |
| Redox Agent | GSH/GSSG |
| Redox Agent Concentration | 3/0.3 mM |
| Refolding Protocol | Lysozyme. The reductive denaturation and the oxidative refolding of lysozyme were carried out as described (Reddy K. et al. 2005). Briefly, the renaturation of the protein was initiated by rapid mixing of reduced denatured lysozyme with 60 volumes of degassed renaturation buffer (3 mM GSH, 0.3 mM GSSG, 1 mM EDTA, 0.1 M Tris/HCl [pH 8.2]), containing organic salts as indicated. The final concentration of lysozyme was 280 μg mL−1. For the screening experiments, the samples were incubated overnight at room temperature and analyzed for lysozyme activity after 18–24 h. Native lysozyme, incubated under the same conditions, without added organic salts, served as control. |
| Refolding Assay | Activity assay |
| Refolding Chaperones | None |
| Refolding Additives | N-substitued N-methylimidazolium cations |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |