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 | Lu D, Liu Z, Zhang M, Wang X, Liu Z (2006) Biochemical Engineering Journal, 27, 336-343 |
| 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 | pH 8.6, 0.1 M Tris–HCl containing 8 M urea, 30 mM DTT and 1 mM EDTA |
| Refolding Buffer | 1.6 M urea, 1 mM GSSG, 4 mM GSH, 1 mM EDTA, and DGP |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.2 |
| Refolding Temperature | 0.0 |
| Protein Concentration | 50 mg/ mL |
| Refolding Time | n/a |
| Redox Agent | GSH/GSSG |
| Redox Agent Concentration | 4/1 mM |
| Refolding Protocol | The denatured lysozyme sample was mixed with pH 8.2, 0.1 M Tris–HCl containing various amounts of GSSG and GSH in the presence or absence of DGP at a specific dilution ratio. The final concentrations of the ingredients in the refolding solution were 1.6 M urea, 1 mM GSSG, 4 mM GSH, 1 mM EDTA, and DGP and lysozyme at their expected concentrations. Then, the solution was stirred constantly at a specified thermostat temperature. The refolding yield was defined as the activity of the refolding solution relative to that of the control containing native lysozyme. |
| Refolding Assay | Kinetic analysis |
| Refolding Chaperones | None |
| Refolding Additives | Dextran-grafted-PNIPAAm (DGP) |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |