LuxS coexpression enhances yields of recombinant proteins in Escherichia coli in part through posttranscriptional control of GroEL.

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TitleLuxS coexpression enhances yields of recombinant proteins in Escherichia coli in part through posttranscriptional control of GroEL.
Publication TypeJournal Article
Year of Publication2011
AuthorsTsao, C-Y, Wang, L, Hashimoto, Y, Yi, H, March, JC, DeLisa, MP, Wood, TK, Valdes, JJ, Bentley, WE
JournalAppl Environ Microbiol
Volume77
Issue6
Pagination2141-52
Date Published2011 Mar
ISSN1098-5336
KeywordsBacterial Proteins, Blotting, Western, Carbon-Sulfur Lyases, Chromatography, High Pressure Liquid, Escherichia coli, Escherichia coli Proteins, Gene Expression Regulation, Bacterial, Heat-Shock Proteins, Homoserine, Lactones, Recombinant Proteins, Reverse Transcriptase Polymerase Chain Reaction
Abstract

Cell-to-cell communication, or quorum sensing (QS), enables cell density-dependent regulation of bacterial gene expression which can be exploited for the autonomous-signal-guided expression of recombinant proteins (C. Y. Tsao, S. Hooshangi, H. C. Wu, J. J. Valdes, and W. E. Bentley, Metab. Eng. 12:291-297, 2010). Earlier observations that the metabolic potential of Escherichia coli is conveyed via the QS signaling molecule autoinducer-2 (AI-2) suggested that the capacity for protein synthesis could also be affected by AI-2 signaling (M. P. DeLisa, J. J. Valdes, and W. E. Bentley, J. Bacteriol. 183:2918-2928, 2001). In this work, we found that simply adding conditioned medium containing high levels of AI-2 at the same time as inducing the synthesis of recombinant proteins doubled the yield of active product. We have hypothesized that AI-2 signaling "conditions" cells as a natural consequence of cell-to-cell communication and that this could tweak the signal transduction cascade to alter the protein synthesis landscape. We inserted luxS (AI-2 synthase) into vectors which cosynthesized proteins of interest (organophosphorus hydrolase [OPH], chloramphenicol acetyltransferase [CAT], or UV-variant green fluorescent protein [GFPuv]) and evaluated the protein expression in luxS-deficient hosts. In this way, we altered the level of luxS in the cells in order to "tune" the synthesis of AI-2. We found conditions in which the protein yield was dramatically increased. Further studies demonstrated coincident upregulation of the chaperone GroEL, which may have facilitated higher yields and is shown for the first time to be positively regulated at the posttranscriptional level by AI-2. This report is the first to demonstrate that the protein synthesis capacity of E. coli can be altered by rewiring quorum sensing circuitry.

DOI10.1128/AEM.02347-10
Alternate JournalAppl. Environ. Microbiol.
PubMed ID21278275
PubMed Central IDPMC3067317