Refolding Record:
| Protein | |
|---|---|
| Protein Name | xylanase |
| Abbreviated Name | XYN4 |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Aspergillus niger |
| UniProt Accession | Q12550 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 168 |
| Molecular Weight | 18131.2 |
| Pi | 6.8 |
| Molecular Weight | 18131.2 |
| Disulphides | 1 |
| Full Sequence |
KVTAAFAGLLVTAFAPPVPEPVLVSRSAGINYVQNYNGNLGDFTYDESAGTFSMYWEDGVSSDFVVGLGWTTGSSNAITYSAEYSASGSSSYLAVYGWVNYLRLNTTSSRITVITTLAARPQALVPSTLMEAPTKSAPTLELTNRPSREQARSRSTSPFERARAHLER
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Shah S, Gupta MN. (2008) Biochim Biophys Acta., 1784, 363-367 |
| 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 | Multi-walled carbon nanotubes |
| Wash Buffer | n/a |
| Solubilization Buffer | 4.8 ml of 0.05 M Tris acetate buffer, pH 9.2, containing 10 M urea (final concentration 8 M) and 100 mM (final concentration 80 mM) dithiothreitol (DT |
| Refolding Buffer | The MWNTs were sonicated in 0.1 ml of 50 mM sodium acetate buffer pH 5.0 |
| Pre-Refolding Purification | None |
| Tag Cleaved | no |
| Refolding pH | 5.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | 0.5 |
| Refolding Time | 30 min |
| Redox Agent | None |
| Redox Agent Concentration | n/a,n/a,n/a,n/a,n/a,n/a,n/a |
| Refolding Protocol | Refolding of xylanase by adsorption on MWNTs The MWNTs were sonicated in 0.1 ml of 50 mM sodium acetate buffer pH 5.0 for 30 min. Denatured xylanase solution was added to these, in a total volume of 0.5 ml. At this stage, the MWNTs were found to form a fine suspension. It has been reported earlier that sonication of SWNTs in the presence of adequate concentration was found to even dissolve SWNTs [14]. The mixture was kept at 25 °C with constant shaking at 200 rpm for 1 h. After incubation, mixture was centrifuged at 10,000 ×g at 25 °C for 10 min. The supernatant was removed. The MWNTs were dispersed in 0.5 ml of acetate buffer pH 5.6. The xylanase activity was measured both in the supernatant and in the bound form in MWNTs. |
| Refolding Assay | Enzyme activity,Unspecified |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | |