Enhanced Soluble Production of Biologically Active Recombinant Human p38 Mitogen-Activated-Protein Kinase (MAPK) in Escherichia coli
Abstract:The conditions were optimized for maximum soluble yield of biologically active recombinant p38α mitogen activated protein kinase (MAPK) vis-a-vis insoluble fraction (inclusion body formation). This study reports a rapid, economical and single step purification process for the overproduction of GST tagged p38α 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 p38α MAPK is more potent (>30%) than a commercially available enzyme.
Document Type: Research Article
Publication date: August 1, 2007
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