title: Heat-shock response transcriptional program enables high-yield and high-quality recombinant protein production in Escherichia coli.
abstract: The biosynthesis of soluble, properly folded recombinant proteins in large quantities from Escherichia coli is desirable for academic research and industrial protein production. The basal E. coli protein homeostasis (proteostasis) network capacity is often insufficient to efficiently fold overexpressed proteins. Herein we demonstrate that a transcriptionally reprogrammed E. coli proteostasis network is generally superior for producing soluble, folded, and functional recombinant proteins. Reprogramming is accomplished by overexpressing a negative feedback deficient heat-shock response transcription factor before and during overexpression of the protein-of-interest. The advantage of transcriptional reprogramming versus simply overexpressing select proteostasis network components (e.g., chaperones and co-chaperones, which has been explored previously) is that a large number of proteostasis network components are upregulated at their evolved stoichiometry, thus maintaining the system capabilities of the proteostasis network that are currently incompletely understood. Transcriptional proteostasis network reprogramming mediated by stress-responsive signaling in the absence of stress should also be useful for protein production in other cells.
authors: Department of Chemistry, ‡Department of Molecular and Experimental Medicine, and §Department of Chemical Physiology, ∥The Skaggs Institute for Chemical Biology, The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
authors: Xin
authors: X
authors: Zhang
authors: Yu
authors: Y
authors: Liu
authors: Joseph C
authors: JC
authors: Genereux
authors: Chandler
authors: C
authors: Nolan
authors: Meha
authors: M
authors: Singh
authors: Jeffery W
authors: JW
authors: Kelly
mesh_terms: D001089:Arabinose / Q000378:metabolism; D018835:Chaperonin 10 / Q000235:genetics / Q000378:metabolism; D018834:Chaperonin 60 / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D025461:Feedback, Physiological; D015964:Gene Expression Regulation, Bacterial; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018869:Heat-Shock Response / Q000235:genetics*; D007613:Kanamycin Resistance / Q000235:genetics; D009154:Mutation; D015202:Protein Engineering / Q000379:methods*; D040901:Proteomics / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D014158:Transcription, Genetic
chemical_list: D018835:Chaperonin 10; D018834:Chaperonin 60; C494635:DnaJ protein, E coli; D029968:Escherichia coli Proteins; C489400:GrpE protein, E coli; D050956:HSP40 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D012808:Sigma Factor; C069774:heat-shock sigma factor 32; D001089:Arabinose