title: 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 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.
authors.affiliation: Department of Biotechnology, Ranbaxy Research Laboratories, R&D -III, Plot 20, Sector-18, Udyog Vihar Industrial Area, Gurgaon, India. sunil.khattar@ranbaxy.com
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Khattar
authors.forename: Pankaj
authors.initials: P
authors.lastname: Gulati
authors.forename: Prabuddha K
authors.initials: PK
authors.lastname: Kundu
authors.forename: Vibhuti
authors.initials: V
authors.lastname: Singh
authors.forename: Usha
authors.initials: U
authors.lastname: Bughani
authors.forename: Malini
authors.initials: M
authors.lastname: Bajpai
authors.forename: Kulvinder S
authors.initials: KS
authors.lastname: Saini
mesh_terms: D003001:Cloning, Molecular; D004789:Enzyme Activation; D004926:Escherichia coli / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D048669:MAP Kinase Kinase 6 / Q000096:biosynthesis; D048308:Mitogen-Activated Protein Kinase 14 / Q000096:biosynthesis* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis; D012995:Solubility
chemical_list: D011994:Recombinant Proteins; D048308:Mitogen-Activated Protein Kinase 14; D048669:MAP Kinase Kinase 6; C482566:MAP2K6 protein, human