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 | Jing-Jing Li, Yong-Dong Liu, Fang-Wei Wang, Guang-Hui Ma, Zhi-Guo Su (2004) J Chromatogr A., 1061, 193-199 |
| 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 | Column refolding: hydrophobic interaction chromatography |
| Wash Buffer | n/a |
| Solubilization Buffer | 6 M guanidine hydrochloride (Gdn-HCl), 100 mM Tris–HCl, 1 mM EDTA, 150 mM dithiothreitol (DTT), pH 8.5 |
| Refolding Buffer | 100 mM Tris, 1 mM EDTA, 3 mM GSH, 0.3 mM GSSG, pH 8.5 |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.5 |
| Refolding Temperature | 0.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | GSH/GSSG |
| Redox Agent Concentration | 3/0.3 mM |
| Refolding Protocol | Refolding bypassing through HIC column One hundred microlitres denatured lysozyme was directly applied to different HIC columns (including Packed Poros PE column (1 ml), Hitrap Butyl Sepharose FF (1 ml), Hitrap Octyl Sepharose FF (1 ml) and Hitrap Phenyl Sepharose HP (1 ml)) respectively pre-equilibrated with refolding buffer (100 mM Tris, 1 mM EDTA, 3 mM GSH, 0.3 mM GSSG, pH 8.5) and then was eluted with the buffer at 0.2 ml/min flow rate on AKTA Purifier. Refolding was also carried out in Packed Poros PE column (128 mm × 10 mm i.d.) with the same buffer. |
| Refolding Assay | enzyme activity,Bradford assay,SEC-HPLC |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | 50.9% |
| Purity | n/a |
| Notes | n/a |