Refolding Record:
| Protein | |
|---|---|
| Protein Name | Hen egg- white lysozyme |
| Abbreviated Name | HEWL |
| SCOP Family | C-type Lysozyme |
| Structure Notes | |
| Organism | Chicken (Gallus gallus) |
| UniProt Accession | P00698 |
| 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 | 126 |
| Molecular Weight | 14471.1 |
| Pi | 10.5 |
| Molecular Weight | 14471.1 |
| Disulphides | 4 |
| Full Sequence |
MKSAVLFLLGIIFLEQCGVRGTLVIRNARCSCISTSRGTIHYKSLKDLKQFAPSPNCNKTEIIATLKNGDQTCLDPDSANVKKLMKEWEKKINQKKKQKRGKKHQKNMKNRKPKTPQSRRRSRKTT
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Mannall GJ, Titchener-Hooker NJ, Dalby PA (2007) Biotechnology and Bioengineering, 97, 1523-1534 |
| 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 | RP-HPLC |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | 8 M GdHCl, 32 mM DTT, 50 mM Tris, 1 mM EDTA pH 8 |
| Solubilization Buffer | 5o mM Tris HCl, 4 mM Cytamine, 1 mM EDTA, (0,0.167,0.47,0.717,0.967,1.967) mM GdHCl pH 7 |
| Refolding Buffer | 5o mM Tris HCl, 4 mM Cytamine, 1 mM EDTA, (0,0.167,0.47,0.717,0.967,1.967) mM GdHCl pH 7 |
| Pre-Refolding Purification | RP-HPLC |
| Tag Cleaved | no tag |
| Refolding pH | 7.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | 2 h |
| Redox Agent | Cystamine |
| Redox Agent Concentration | 4 mM |
| Refolding Protocol | Denatured reduced lysozyme (16 mg mL􏰁1) was diluted 1:15 in various refold buffers (six refold buffers in total as detailed in Table I) at ambient temperature and mixed vigorously for 30 s, and left at ambient temperature. After 2 h, a sample (2 mL) was taken and quenched with 10% v/v TFA (200 umL). A further sample was taken at 24 h and left unquenched. Samples were assayed for native lysozyme using RP-HPLC. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |