Skip to main content
Log in

Stability of Escherichia coli strains harboring recombinant plasmids for L-threonine production

  • Published:
Antonie van Leeuwenhoek Aims and scope Submit manuscript

Abstract

Escherichia coli recombinant strains bearing the thr operon have been previously selected for threonine production and phenotypically classified according to antibiotic resistance properties (Nudel et al. 1987).

Further analysis of those strains permitted the isolation and restriction mapping of two different plasmids of 13 kb and 18.6 kb. The smaller one, which expressed tetracycline resistance gave better results on threonine accumulation but it was rather unstable when grown without antibiotic pressure. Therefore, other hosts were transformed with those plasmids to improve stability.

A threonine-auxotrophic strain was a better host for plasmid maintenance and expression of thr operon. Host influence in plasmid-mediated threonine production was studied in terms of specific yields (the ratios of threonine accumulated to biomass values) and of plasmid maintenance (percent of AprTcr clones after cultivation in non selective media).

We also determined that semisynthetic media of defined composition were better than rich media for threonine expression, due to feed-back controls exerted by undesired catabolites accumulated in complex media.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Dagert M & Ehrlich SD (1979) Prolonged incubation in calcium chloride improves the competence of E. coli. Gene 6: 23–28

    Google Scholar 

  • Imanaka T & Aiba S (1981) A perspective on the application of genetic engineering: stability of recombinant plasmid. Ann. N.Y. Acad. Sci. 369: 1–14

    Google Scholar 

  • Joshi S & Yamazaki H (1987) Stability of plasmid pBR322 in E. coli immobilized on cotton cloth during use as resident inoculum. Biotech. Lett. 9: 825–830

    Google Scholar 

  • Maniatis T, Fritsch EF & Sambrook J (1982) Molecular Cloning. A laboratory manual. Cold Spring Harbor Laboratory

  • Miwa K, Tsuchida T, Kurahashi O, Nakamori S, Sano K & Momose H (1980) Threonine production by fermentation. U.K. Patent Application 2049670

  • Miwa K, Nakamori S, Sano K & Momose H (1984) Stability of recombinant plasmids carrying the threonine operon in E. coli. Agric. Biol. Chem. 48: 2233–2237

    Google Scholar 

  • Momose H (1983) New genetic approaches to amino-acid producing strains. In: Developments in Industrial Microbiol. 24: 109–119

  • Noack D, Roth M, Geuther R, Müller G, Undisz K, Hoffmeier Ch & Gáspár S (1981) Maintenance and genetic stability of vector plasmids pBR322 and pBR325 in E. coli K12 strains grown in a chemostat. Molec. Gen. Genet. 184: 121–124

    Google Scholar 

  • Nudel BC, Corradini C, Fraile E & Giulietti AM (1987) Plasmid maintenance and threonine production by recombinant E. coli strains. Biotech. Lett. 9: 77–82

    Google Scholar 

  • Robinson M, Marquardt R, Theu J, McChesney J, Neatherway H, Wöhner G, Deger HM & Prave P (1987) Cloning of the gene coding for aromatic amino acid aminotranferase from an E. coli B Strain. Biotech. Lett. 9: 673–679

    Google Scholar 

  • Ryu DDY & Siegel R (1986) Scale-up of fermentation processes using recombinant organisms. Ann. N.Y. Acad. Sci. 469: 73–82

    Google Scholar 

  • Summers DK & Sherrat DJ (1984) Multimerization of high copy number plasmids causes instability: ColE1 encodes a determinant essential for plasmid monomerization and stability. Cell 36: 1097–1103

    Google Scholar 

  • Yukawa H, Kurusu Y, Shimazu M, Yamagata H & Teresawa M (1988) Stabilization of an E. coli plasmid by a mini-F fragment of DNA. J. Ind. Microbiol. 2: 323–328

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nudel, B.C., Pueyo, M.G., Judewicz, N.D. et al. Stability of Escherichia coli strains harboring recombinant plasmids for L-threonine production. Antonie van Leeuwenhoek 56, 273–282 (1989). https://doi.org/10.1007/BF00418939

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00418939

Key words

Navigation