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Increase of organic solvent tolerance by overexpression of manXYZ in Escherichia coli

  • Mina Okochi
  • Masaki Kurimoto
  • Kazunori Shimizu
  • Hiroyuki HondaEmail author
Applied Genetics and Molecular Biotechnology

Abstract

Transcriptional analysis has been investigated to detect the genes involved in organic solvent tolerance. A time course of gene expression profiles of Escherichia coli after exposure to organic solvents revealed that the expression levels of manX, manY, and manZ genes were strongly upregulated by 13.2-, 10.0-, and 7.0-folds, respectively, after 2 min and then decreased after 10 min. Organic solvent tolerance of E. coli was investigated by inducing overexpression of manX, manY, and manZ genes and manXYZ operon that encode a sugar transporter of the phosphotransferase system. Although the expression of manX, manY, and manZ alone was not effective, the organic solvent tolerance level was increased by the expression of manXYZ. The intracellular hexane level was kept lower in E. coli cells overexpressing manXYZ after 4 h of incubation with hexane.

Keywords

Organic solvent tolerance DNA microarray manXYZ 

Notes

Acknowledgements

This study was carried out as a part of “The Project for Development of a Technological Infrastructure for Industrial Bioprocesses on R&D of New Industrial Science and Technology Frontiers” by the Ministry of Economy, Trade, and Industry (METI) and entrusted by the New Energy and Industrial Technology Development Organization (NEDO). This study was partially supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (no. 17560688).

References

  1. Aono R, Kobayashi H (1997) Cell surface properties of organic solvent-tolerant mutants of Escherichia coli K-12. Appl Environ Microbiol 63:3637–3642Google Scholar
  2. Aono R, Aibe A, Inoue A, Horikoshi K (1991) Preparation of organic solvent tolerant mutants from Escherichia coli K-12. Agric Biol Chem 55:1935–938Google Scholar
  3. Aono R, Negishi T, Nakajima H (1994) Cloning of organic solvent tolerance gene ostA that determines n-hexane tolerance level in Escherichia coli. Appl Environ Microbiol 60:4624–4626Google Scholar
  4. Asako H, Nakajima H, Kobayashi K, Kobayashi M, Aono R (1997) Organic solvent tolerance and antibiotic resistance increased by overexpression of marA in Escherichia coli. Appl Environ Microbiol 63:1428–1433Google Scholar
  5. Bos MP, Tefsen B, Tommassen J (2004) Identification of an outer membrane protein required for the transport of lipopolysaccharide to the bacterial cell surface. Proc Natl Acad Sci USA 101:9417–9422CrossRefGoogle Scholar
  6. Hansch C, Fujita T (1964) ρ–σ–π–Analysis. A method for the correlation of biological activity and chemical structure. J Am Chem Soc 86:1616–1626CrossRefGoogle Scholar
  7. Hayashi S, Aono R, Hanai T, Mori H, Kobayashi T, Honda H (2003) Analysis of organic solvent tolerance in Escherichia coli using gene expression profiles from DNA microarrays. J Biosci Bioeng 95:379–383Google Scholar
  8. Heipieper HJ, Weber FJ, Sikkema J, Keweloh H, de Bont JAM (1994) Mechanisms of resistance of whole cells to toxic organic solvents. Trends Biotechnol 12:409–414CrossRefGoogle Scholar
  9. Ingram LO (1977) Changes in lipid composition of Escherichia coli resulting from growth with organic solvents and food additives. Appl Environ Microbiol 33:1233–1236Google Scholar
  10. Inoue A, Horikoshi K (1989) A Pseudomonas thrives in high concentrations of toluene. Nature 338:264–265CrossRefGoogle Scholar
  11. Isken S, de Bont JAB (1996) Active efflux of toluene in a solvent-resistant bacterium. J Bacteriol 178:6056–6058Google Scholar
  12. Kingsman AJ, Clarke L, Mortimer RK, Carbon J (1979) Replication in Saccharomyces cerevisiae of plasmid pBR313 carrying DNA from the yeast trp1 region. Gene 7:141–152CrossRefGoogle Scholar
  13. Li L, Komatsu T, Inoue A, Horikoshi K (1995) A toluene-tolerant mutant of Pseudomonas aeruginosa lacking the outer membrane protein F. Biosci Biotechnol Biochem 59:2358–2359CrossRefGoogle Scholar
  14. Matsui K, Hirayama T, Kuroda K, Shirahige K, Ashikari T, Ueda M (2006) Screening for candidate genes involved in tolerance to organic solvents in yeast. Appl Microbiol Biotechnol 71:75–79CrossRefGoogle Scholar
  15. Miura S, Zou W, Ueda M, Tanaka A (2000) Screening of genes involved in isooctane tolerance in Saccharomyces cerevisiae by using mRNA differential display. Appl Environ Microbiol 66:4883–4889CrossRefGoogle Scholar
  16. Nakajima H, Kobayashi K, Kobayashi M, Asako H, Aono R (1995) Overexpression of the robA gene increases organic solvent tolerance and multiple antibiotic and heavy metal ion resistance in Escherichia coli. Appl Environ Microbiol 61:2302–2307Google Scholar
  17. Pinkart HC, Wolfram JW, Rogers R, White DC (1996) Cell envelope changes in solvent-tolerant and solvent-sensitive Pseudomonas putida strains following exposure to o-xylene. Appl Environ Microbiol 62:1129–1132Google Scholar
  18. Rosenberg M, Gutnick D, Rosenberg E (1980) Adherence of bacteria to hydrocarbons: a simple method for measuring cell-surface hydrophobicity. FEMS Lett 9:29–33CrossRefGoogle Scholar
  19. Segura A, Rojas A, Hurtado A, Ramos JL, Heurtas M-J (2003) Comparative genemic analysis of solvent extrusion pumps in Pseudomonas strains exhibiting different degrees of solvent tolerance. Extremophiles 7:371–376CrossRefGoogle Scholar
  20. Shimizu K, Hayashi S, Doukyu N, Kobayashi T, Honda H (2005a) Time-course data analysis of gene expression profiles reveals purR regulon concerns in organic solvent tolerance in Escherichia coli. J Biosci Bioeng 99:72–74CrossRefGoogle Scholar
  21. Shimizu K, Hayashi S, Kako T, Suzuki M, Tsukagoshi N, Doukyu N, Kobayashi T, Honda H (2005b) Discovery of glpC as an organic solvent tolerance related gene in Escherichia coli using gene expression profiles from DNA microarrays. Appl Environ Microbiol 71:1093–1096CrossRefGoogle Scholar
  22. Sikkema J, de Bont JAM, Poolman B (1995) Mechanisms of membrane toxicity of hydrocarbons. Microbiol Rev 59:201–222Google Scholar
  23. Tsugakoshi N, Aono R (2000) Entry into and release of solvents by Escherichia coli in an organic-aqueous two-liquid phase system and substrate specificity of the AcrAB-TolC solvent-extruding pump. J Bacteriol 182:4803–4810CrossRefGoogle Scholar
  24. Zhang W, Needham DL, Coffin M, Rooker A, Hurban P, Tanzer MM, Shuster JR (2003) Microarray analysis of the metabolic responses of Saccharomyces cerevisiae to organic solvent dimethyl sulfoxide. J Ind Microbiol Biotechnol 30:57–69Google Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Mina Okochi
    • 1
  • Masaki Kurimoto
    • 1
  • Kazunori Shimizu
    • 1
  • Hiroyuki Honda
    • 1
    Email author
  1. 1.Department of BiotechnologySchool of Engineering, Nagoya UniversityNagoyaJapan

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