Enhanced Soluble Production of Biologically Active Recombinant Human p38 Mitogen-Activated-Protein Kinase (MAPK) in Escherichia coli

Title: Enhanced Soluble Production of Biologically Active Recombinant Human p38 Mitogen-Activated-Protein Kinase (MAPK) in Escherichia coli

Volume: 14 Issue: 8

Author(s): Sunil K. Khattar, Pankaj Gulati, Prabuddha K. Kundu, Vibhuti Singh, Usha Bughani, Malini Bajpai and Kulvinder. S. Saini

Affiliation:

Keywords: Affinity chromatography, enzyme kinetics, GST tag, MKK6, phosphorylation, soluble p38α MAPK

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.

Cite this article as:

Sunil K. Khattar , Pankaj Gulati , Prabuddha K. Kundu , Vibhuti Singh , Usha Bughani , Malini Bajpai and Kulvinder. S. Saini , Enhanced Soluble Production of Biologically Active Recombinant Human p38 Mitogen-Activated-Protein Kinase (MAPK) in Escherichia coli, Protein & Peptide Letters 2007; 14 (8) . https://dx.doi.org/10.2174/092986607781483660

DOI https://dx.doi.org/10.2174/092986607781483660	Print ISSN 0929-8665
Publisher Name Bentham Science Publisher	Online ISSN 1875-5305