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, Zhang K, Liu Z (2005) Biochemical Engineering Journal, 25, 141-149 |
| 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 | 0.1 M Tris–HCl, pH 8.2, containing 20 mM CTAB, and further diluted with 0.1 M, pH 8.2 Tris–HCl containing 0.4 mM GSSG, 4 mM GSH and 1 mM EDTA as well as B-CD-g-PNIPAAm or B-CD |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.2 |
| Refolding Temperature | 0.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | GSH/GSSG |
| Redox Agent Concentration | 4/0.4 mM |
| Refolding Protocol | The two-step dilution method was used for lysozyme refolding. In the first step, a denatured lysozyme sample of 50 mg/ml was mixed with 0.1 M Tris–HCl, pH 8.2, containing 20 mM CTAB, to give a specific mole ratio of CTAB to lysozyme. The mixture was incubated at room temperature for 30 min. In the second step, the solution was further diluted by 0.1 M, pH 8.2 Tris–HCl containing 0.4 mM GSSG, 4 mM GSH and 1 mM EDTA as well as B-CD-g-PNIPAAm or B-CD of specified concentration. The refolding yield was defined as the activity of the refolding solution relative to that of the control containing native lysozyme. |
| Refolding Assay | Fluorescence,Protein activity assay |
| Refolding Chaperones | None |
| Refolding Additives | Dextran-grafted-PNIPAAm (DGP) |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |