Enhanced soluble production of biologically active recombinant human p38 mitogen-activated-protein kinase (MAPK) in Escherichia coli
| Autor: | Sunil K. Khattar, Kulvinder Singh Saini, Vibhuti Singh, Prabuddha K. Kundu, Malini Bajpai, Pankaj Gulati, Usha Bughani |
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| Rok vydání: | 2007 |
| Předmět: |
MAPK/ERK pathway
MAP Kinase Kinase 6 medicine.disease_cause Biochemistry law.invention Mitogen-Activated Protein Kinase 14 Structural Biology law medicine Escherichia coli Humans Cloning Molecular chemistry.chemical_classification biology Kinase Biological activity General Medicine Molecular biology Recombinant Proteins Enzyme Activation Kinetics Enzyme chemistry Solubility Mitogen-activated protein kinase biology.protein Recombinant DNA Phosphorylation |
| Zdroj: | Protein and peptide letters. 14(8) |
| ISSN: | 0929-8665 |
| Popis: | The conditions were optimized for maximum soluble yield of biologically active recombinant p38alpha mitogen activated protein kinase (MAPK) vis-à-vis insoluble fraction (inclusion body formation). This study reports a rapid, economical and single step purification process for the overproduction of GST tagged p38alpha MAPK. A yield of 18 mg of highly purified and soluble protein per liter of bacterial culture within 6 h timeframe was achieved. The purified protein was found to be biologically suitable for phosphorylation by upstream kinases and was catalytically active. We further demonstrated that our in-house p38alpha MAPK is more potent (30%) than a commercially available enzyme. |
| Databáze: | OpenAIRE |
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