title: GroEL-GroES assisted folding of multiple recombinant proteins simultaneously over-expressed in Escherichia coli.
abstract: Folding of aggregation prone recombinant proteins through co-expression of chaperonin GroEL and GroES has been a popular practice in the effort to optimize preparation of functional protein in Escherichia coli. Considering the demand for functional recombinant protein products, it is desirable to apply the chaperone assisted protein folding strategy for enhancing the yield of properly folded protein. Toward the same direction, it is also worth attempting folding of multiple recombinant proteins simultaneously over-expressed in E. coli through the assistance of co-expressed GroEL-ES. The genesis of this thinking was originated from the fact that cellular GroEL and GroES assist in the folding of several endogenous proteins expressed in the bacterial cell. Here we present the experimental findings from our study on co-expressed GroEL-GroES assisted folding of simultaneously over-expressed proteins maltodextrin glucosidase (MalZ) and yeast mitochondrial aconitase (mAco). Both proteins mentioned here are relatively larger and aggregation prone, mostly form inclusion bodies, and undergo GroEL-ES assisted folding in E. coli cells during over-expression. It has been reported that the relative yield of properly folded functional forms of MalZ and mAco with the exogenous GroEL-ES assistance were comparable with the results when these proteins were overexpressed alone. This observation is quite promising and highlights the fact that GroEL and GroES can assist in the folding of multiple substrate proteins simultaneously when over-expressed in E. coli. This method might be a potential tool for enhanced production of multiple functional recombinant proteins simultaneously in E. coli.
authors.affiliation: Kusuma School of Biological Sciences, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.
authors.forename: Megha
authors.initials: M
authors.lastname: Goyal
authors.affiliation: Kusuma School of Biological Sciences, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India. Electronic address: tkchaudhuri@bioschool.iitd.ac.in.
authors.forename: Tapan K
authors.initials: TK
authors.lastname: Chaudhuri
mesh_terms: D000154:Aconitate Hydratase / Q000096:biosynthesis / Q000235:genetics; D018835:Chaperonin 10 / Q000502:physiology*; D018834:Chaperonin 60 / Q000502:physiology*; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000235:genetics; D006026:Glycoside Hydrolases / Q000096:biosynthesis / Q000235:genetics; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D015533:Transcriptional Activation
chemical_list: D018835:Chaperonin 10; D018834:Chaperonin 60; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; C071233:malZ protein, E coli; D000154:Aconitate Hydratase
keywords: Aconitase; Chaperone assisted protein folding; GroEL; GroES; Maltodextrin glucosidase; Multiple protein folding in Escherichia coli