Refolding Record:
| Protein | |
|---|---|
| Protein Name | beta-lactamase |
| Abbreviated Name | beta-lactamase |
| SCOP Family | beta-Lactamase/D-ala carboxypeptidase |
| Structure Notes | |
| Organism | Escherichia coli |
| UniProt Accession | P00811 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Multi-domain proteins (alpha and beta) |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 358 |
| Molecular Weight | 39551.1 |
| Pi | 8.7837 |
| Molecular Weight | 39551.1 |
| Disulphides | 0 |
| Full Sequence |
APQQINDIVHRTITPLIEQQKIPGMAVAVIYQGKPYYFTWGYADIAKKQPVTQQTLFELGSVSKTFTGVLGGDAIARGEIKLSDPTTKYWPELTAKQWNGITLLHLATYTAGGLPLQVPDEVKSSSDLLRFYQNWQPAWAPGTQRLYANSSIGLFGALAVKPSGLSFEQAMQTRVFQPLKLNHTWINVPPAEEKNYAWGYREGKAVHVSPGALDAEAYGVKSTIEDMARWVQSNLKPLDINEKTLQQGIQLAQSRYWQTGDMYQGLGWEMLDWPVNPDSIINGSDNKIALAARPVKAITPPTPAVRASWVHKTGATGGFGSYVAFIPEKELGIVMLANKNYPNPARVDAAWQILNALQ
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | St.John, Richard, J. (1999) Proc. Natl. Acad. Sci. USA, 96, 13029-13033 |
| Project Aim | Folding |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | RB791 |
| Expression Temp | 37.0 |
| Expression Time | 18 hours |
| Expression Vector | pGB1 |
| Expression Protocol | Unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Microfluidizer |
| Lytic Agent | Lysozyme |
| Pre-Refolding Purification | Washing inclusion body |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | High Pressure |
| Wash Buffer | 50 mM Tris pH 8.0, 0.2% lysozyme |
| Solubilization Buffer | Unknown |
| Refolding Buffer | Unknown |
| Pre-Refolding Purification | Washing inclusion body |
| Tag Cleaved | no tag |
| Refolding pH | 7.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | 48 hrs |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | |
| Refolding Yield | 85 |
| Purity | |
| Notes | |