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Control of Acetate Production Rate in Escherichia coli by Regulating Expression of Single-Copy pta Using $lacI^Q$ in Multicopy Plasmid  

Lee, Sun-Gu (Department of Chemical and Biochemical Engineering, Pusan National University)
Liao, James C (Department of Chemical and Biomolecular Engineering, University of California Los Angeles)
Publication Information
Journal of Microbiology and Biotechnology / v.18, no.2, 2008 , pp. 334-337 More about this Journal
Abstract
A tightly regulated gene expression system composed of a single-copy target gene under the control of a lac promoter derivative and lacI gene in a multicopy plasmid is proposed, and its ability to control the flux of a metabolic pathway is demonstrated. A model system to control the flux of acetyl-CoA to acetyl phosphate was constructed by integrating pta, a gene encoding phosphotransacetylase, under a tac promoter into the chromosome of E. coli with a pta-negative background and transforming a multicopy plasmid containing the $lacI^Q$ gene into the strain. The production rate of acetate was shown to be tightly controlled when varying the concentration of the inducer (IPTG) in he model system.
Keywords
Metabolic engineering; tight regulation; metabolic flux; tac promoter; lacI;
Citations & Related Records
Times Cited By KSCI : 4  (Citation Analysis)
Times Cited By Web Of Science : 1  (Related Records In Web of Science)
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1 Keasling, J. D. 1999. Gene-expression tools for the metabolic engineering of bacteria. TIBTECH 17: 452-460   DOI   ScienceOn
2 Stephanopoulos, G. 1999. Metabolic fluxes and metabolic engineering. Metab. Eng. 1: 1-11   DOI   ScienceOn
3 Stephanopoulos, G. N., A. A. Aristidou, and J. Nielsen. 1998. Metabolic Engineering. Academic Press
4 Sorensen, H. P. and K. K. Mortensen. 2005. Advanced genetic strategies for recombinant protein expression in E. coli. J. Biotechnol. 115: 113-128   DOI   ScienceOn
5 Haldimann, A. and B. L. Wanner. 2001. Conditional-replication, integration, excision, and retrieval plasmid-host systems for gene structure-function studies of bacteria. J. Bacteriol. 183: 6384-6393   DOI   ScienceOn
6 Kim, J. Y. H. and H. J. Cha. 2003. Down-regulation of acetate pathway through antisense strategy in Escherichia coli: Improved foreign protein production. Biotechnol. Bioeng. 83: 841-853   DOI   ScienceOn
7 Raab, R. M., K. Tyo, and G. Stephanopoulos. 2005. Metabolic engineering. Adv. Biochem. Eng. Biotechnol. 100: 1-17
8 Bowers, L. M., K. LaPointa, L. Anthony, A. Pluciennik, and M. Filutowicz. 2004. Bacterial expression system with tightly regulated gene expression and plasmid copy number. Gene 340: 11-18   DOI   ScienceOn
9 Furste, J. P., W. Pansegrau, R. Frank, H. Blocker, P. Scholz, M. Bagdasarian, and E. Lanka. 1986. Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene 48: 119-131   DOI   ScienceOn
10 Lee, S. G., Y. J. Kim, S. I. Han, Y.-K. Oh, S. Park, Y. H. Kim, and K.-S. Hwang. 2006. Simulation of dynamic behavior of glucose- and tryptophan-grown Escherichia coli using constraintbased metabolic models with a hierarchical regulatory network. J. Microbiol. Biotechnol. 16: 993-998   과학기술학회마을
11 Lutz, R. and H. Bujard. 1997. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and $AraC/I_{1}-I_{2}$ regulatory elements. Nucleic Acids Res. 25: 1203-1210   DOI   ScienceOn
12 Nakano, K., M. Rischke, S. Sato, and H. Markl. 1997. Influence of acetic acid on the growth of Escherichia coli K12 during high-cell-density cultivation in a dialysis reactor. Appl. Microbiol. Biotechnol. 48: 597-601   DOI   ScienceOn
13 Kim, T.-Y. and S.-Y. Lee. 2006. Accurate metabolic flux analysis through data reconciliation of isotope balance-based data. J. Microbiol. Biotechnol. 16: 1139-1143   과학기술학회마을
14 Fung, E., W. W. Wong, J. K. Suen, T. Bulter, S. Lee, and J. C. Liao. 2005. A synthetic gene-metabolic oscillator. Nature 435: 118-122   DOI   ScienceOn
15 Asenjo, J. A., P. Ramirez, I. Rapaport, J. Aracena, E. Goles, and B. A. Andrews. 2007. A discrete mathematical model applied to genetic regulation and metabolic networks. J. Microbiol. Biotechnol. 17: 496-510   과학기술학회마을
16 Bulter, T., S. G. Lee, W. W. Wong, E. Fung, M. R. Connor, and J. C. Liao. 2004. Design of artificial cell-cell communication using gene and metabolic network. Proc. Natl. Acad. Sci. USA 101: 2299-2304
17 Baneyx, F. 1999. Recombinant protein expression in Escherichia coli. Curr. Opin. Biotechnol. 10: 411-421   DOI   ScienceOn
18 Farmer, W. R. and J. C. Liao. 1997. Reduction of aerobic acetate production by Escherichia coli. Appl. Environ. Microbiol. 63: 3205-3210
19 Glascock, C. B. and M. J. Weickert. 1998. Using $chromosomal lacI^{Q1}$ to control expression of genes on high-copy-number plasmids in Escherichia coli. Gene 223: 221-231   DOI
20 Jones, K. L., S. Kim, and J. D. Keasling. 2000. Low-copy plasmids can perform as well as or better than high-copy plasmids for metabolic engineering of bacteria. Metabolic Eng. 2: 328-338   DOI   ScienceOn
21 Oh, M.-K., M.-J. Cha, S.-G. Lee, L. Rohlin, and J. C. Liao. 2006. Dynamic gene expression profiling of Escherichia coli in carbon source transition from glucose to acetate. J. Microbiol. Biotechnol. 16: 543-549   과학기술학회마을