Refolding Record:
| Protein | |
|---|---|
| Protein Name | Arginine Deiminase |
| Abbreviated Name | AD |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Mycoplasma arginini |
| UniProt Accession | P23793 |
| 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 | 416 |
| Molecular Weight | 46507.3 |
| Pi | 5.23093 |
| Molecular Weight | 46507.3 |
| Disulphides | Unknown |
| Full Sequence |
MSVFDSKFKGIHVYSEIGELESVLVHEPGREIDYITPARLDELLFSAILESHDARKEHKQ
FVAELKANDINVVELIDLVAETYDLASQEAKDKLIEEFLEDSEPVLSEEHKVVVRNFLKA
KKTSRELVEIMMAGITKYDLGIEADHELIVDPMPNLYFTRDPFASVGNGVTIHYMRYKVR
QRETLFSRFVFSNHPKLINTPWYYDPSLKLSIEGGDVFIYNNDTLVVGVSERTDLQTVTL
LAKNIVANKECEFKRIVAINVPKWTNLMHLDTWLTMLDKDKFLYSPIANDVFKFWDYDLV
NGGAEPQPVENGLPLEGLLQSIINKKPVLIPIAGEGASQMEIERETHFDGTNYLAIRPGV
VIGYSRNEKTNAALEAAGIKVLPFHGNQLSLGMGNARCMSMPLSRKDVKW
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Misawa S, Aoshima M, Takaku H, Matsumoto M, Hayashi H. (1994) J Biotechnology, 36, 145-155 |
| Project Aim | Structure-Function |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | JM101 |
| Expression Temp | 37.0 |
| Expression Time | 24h |
| Expression Vector | pMK2 |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | 10mM potassium phosphate, 1mM EDTA, 4% Triton X-100, pH 7.0 |
| Solubilization Buffer | 50mM Tris-HCl, 10mM DTT, 6M guanidinium chloride, pH 8.5 |
| Refolding Buffer | 10mM potassium phosphate |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.5 |
| Refolding Temperature | 15.0 |
| Protein Concentration | |
| Refolding Time | 45h |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The solubilized inclusion bodies were rapidly diluted with 20L of 10mM potassium phosphate, pH 7.0 and stirred constantly at room temperature for 45h. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 70mg/L culture |
| Purity | |
| Notes | |