Refolding Record:
| Protein | |
|---|---|
| Protein Name | Manganese peroxidase |
| Abbreviated Name | MN peroxidase |
| SCOP Family | CCP-like |
| Structure Notes | |
| Organism | Phanerochaete chrysosporium |
| UniProt Accession | Q12170 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 388 |
| Molecular Weight | 40142.3 |
| Pi | 4.55236 |
| Molecular Weight | 40142.3 |
| Disulphides | 5 |
| Full Sequence |
MAFGSLLAFVALAAITRAAPTAESAVCPDGTRVTNAACCAFIPLAQDLQETLFQGDCGED
AHEVIRLTFHDAIAISQSLGPQAGGGADGSMLHFPTIEPNFSANNGIDDSVNNLLPFMQK
HDTISAADLVQFAGAVALSNCPGAPRLEFMAGRPNTTIPAVEGLIPEPQDSVTKILQRFE
DAGNFSPFEVVSLLASHTVARADKVDETIDAAPFDSTPFTFDTQVFLEVLLKGTGFPGSN
NNTGEVMSPLPLGSGSDTGEMRLQSDFALARDERTACFWQSFVNEQEFMAASFKAAMAKL
AILGHSRSSLIDCSDVVPVPKPAVNKPATFPATKGPKDLDTLTCKALKFPTLTSDPGATE
TLIPHCSNGGMSCPGVQFDGPA
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Whitwam RE, Gazarian IG, Tien M. (1995) Biochemical and Biophysical Research Com, 216, 1013-1017 |
| Project Aim | Functional Studies |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21 |
| Expression Temp | 37.0 |
| Expression Time | 3h |
| Expression Vector | pET21a |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | Lysozyme |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | unknown |
| Solubilization Buffer | 2mM EDTA, 1mM DTT, 8M urea, 50mM Tris-HCl, pH 8.0 |
| Refolding Buffer | 5mM CaCl2, 0.7mM GSH, 5 micromolar hemin, 50mM Tris-HCl, 2M urea |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 4.0 |
| Protein Concentration | |
| Refolding Time | 16h |
| Redox Agent | GSH |
| Redox Agent Concentration | n/a,n/a |
| Refolding Protocol | The inclusion body pellet was solubilized in 2mM EDTA, 1mM DTT, 8M urea, 50mM Tris-HCl, pH 8.0, and refolded by rapid dilution yielding a solution containing 5mM CaCl2, 0.7mM GST, 5 micromolar hemin, 50mM Tris-HCl, 2M urea. The protein was allowed to refold overnight at 4 degrees C. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | |
| Notes | |