1887

Abstract

The evolution of MG1655 mutants was followed over 126 d in a glycerol-limited chemostat at a dilution rate of 0.05 h. This corresponds to a total of 217 generations at a doubling time of 13.9 h. After this time, nearly 90% of the chemostat population consisted of evolved mutant strains as determined by their altered colony morphologies on plates. Two mutants were isolated that exhibited generally improved growth phenotypes in batch cultivations on glycerol, glucose or the gluconeogenic substrate acetate. Higher specific growth rates and increased biomass yields were found for both mutants. For one mutant, this behaviour was combined with significantly reduced secretion of overflow metabolites when either glycerol or glucose was the carbon source. Additionally, during all growth phases of a batch cultivation, this mutant exhibited increased resistance to a variety of adverse conditions including heat shock, osmotic stress and nutrient deprivation. It also displayed significantly shorter lag phases.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-5-1567
1997-05-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/5/mic-143-5-1567.html?itemId=/content/journal/micro/10.1099/00221287-143-5-1567&mimeType=html&fmt=ahah

References

  1. Bachmann B. J. 1987; Derivations and genotypes of some mutant derivatives of Escherichia coli K-12. In Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology, Edited by F. C. Neidhardt, J. L. Ingraham, K. Brooks Low, B. Magasanik, M. Schaechter & H. E. Umbarger. Washington, DC:. American Society for Microbiology1190–1219.
    [Google Scholar]
  2. Calhoun M. W., Oden K. L., Gennis R. B., Teixeira de Mattos M. J., Neijssel O. M. 1993; Energetic efficiency of Escherichia coli: effects of mutations in components of the aerobic respiratory chain. J Bacteriol 175:3020–3025.
    [Google Scholar]
  3. Cheng C. Y., Yabe I., Toda K. 1989; Predominant growth of alpha-amylase regulation mutant in continuous culture of Bacillus caldolyticus. J Ferment Bioeng 67:176–181.
    [Google Scholar]
  4. Chesbro W., Arbige M., Eifert R. 1990; When nutrient limitation places bacteria in the domains of slow growth: metabolic, morphologic and cell cycle behavior. FEMS Microbiol Ecol 74:103–119.
    [Google Scholar]
  5. Chou C.-H., Bennett G. N., San K.-Y. 1994; Effect of modified glucose uptake using genetic engineering techniques on high-level recombinant protein production in Escherichia coli dense cultures. Biotechnol Bioeng 44:952–960.
    [Google Scholar]
  6. Contois D. E. 1959; Kinetics of bacterial growth: relationship between population density and specific growth rate in continuous cultures. J Gen Microbiol 21:40–50.
    [Google Scholar]
  7. Dedhia N. N., Hottinger T., Bailey J. E. 1994; Overproduction of glycogen in Escherichia coli blocked in the acetate pathway improves cell-growth. Biotechnol Bioeng 44:132–139.
    [Google Scholar]
  8. Dykhuizen D., Hartl D. 1981; Evolution of competitive ability in Escherichia coli. Evolution 35:581–594.
    [Google Scholar]
  9. Dykhuizen D. E., Hartl D. L. 1983; Selection in chemostats. Microbiol Rev 47:150–168.
    [Google Scholar]
  10. Helling R. B., Vargas C. N., Adams J. 1987; Evolution of Escherichia coli during growth in a constant environment. Genetics 116:349–358.
    [Google Scholar]
  11. Kakuda H., Shiroishi K., Hosono K., Ichihara S. 1994; Construction of a Pta-Ack pathway deletion mutant of Escherichia coli and characteristic growth profiles of the mutants in a rich medium. Biosci Biotechnol Biochem 58:2232–2235.
    [Google Scholar]
  12. Kubitschek H. E. 1970; Zntroduction to Research with Continuous Cultures. Englewood-Cliffs:. Prentice-Hall.
    [Google Scholar]
  13. Kurland C. G. 1992; Translational accuracy and the fitness of bacteria. Annu Rev Genet 26:29–50
    [Google Scholar]
  14. MacPhee D. G. 1993; Directed mutations: a critical analysis. ASM News 59:297–299
    [Google Scholar]
  15. Matin A. 1992; Physiology, molecular biology and applications of the bacterial starvation response. J Appl Bacteriol Symp (Suppl.) 73:49S–57S
    [Google Scholar]
  16. Matin A., Auger E. A., Blum P. H., Schultz J. E. 1989; Genetic basis of starvation survival in nondifferentiating bacteria. Annu Rev Microbiof 43:293–316
    [Google Scholar]
  17. Moser H. 1958; The Dynamics of Bacterial Populations Maintained in the Chemostat. Washington, DC:. Carnegie Institute of Washington (Publication 61).
    [Google Scholar]
  18. Notley L., Ferenci T. 1996; Induction of RhoS-dependent functions in glucose-limited continuous culture: what level of nutrient limitation induces the stationary phase of Escherichia coli. J Bacteriol 178:1465–1468
    [Google Scholar]
  19. Novick A., Szilard L. 1950; Experiments with the chemostat on spontaneous mutations in bacteria. Proc Natl Acad Sci USA 36:708–719
    [Google Scholar]
  20. Paquin C ., Adams J. 1983; Frequency of fixation of adaptive mutations is higher in evolving diploid than haploid yeast populations. Nature 302:495–500
    [Google Scholar]
  21. Rosenzweig R. F., Sharp R. R., Treves D. S., Adams J. 1994; Microbial evolution in a simple unstructured environment: genetic differentiation in Escherichia coli. Genetics 137:1–13
    [Google Scholar]
  22. Sakamoto S., Terada I., Lee Y. C., Uehara K., Matsuzawa H., Ijima M. 1996; Efficient production of Thermus protease aqualysin-1 in Escherichia coli: effect of cloned gene structure and 2-stage culture. Appl Microbiol Biotechnol 45:94–101
    [Google Scholar]
  23. Svitil A. L., Cashel M., Zyskind J. W. 1993; Guanosine tetraphosphate inhibits protein synthesis in vivo. J Biol Chem 268:2307–2311
    [Google Scholar]
  24. Tedin K., Bremer H. 1992; Toxic effects of high levels of ppGpp in Escherichia coli are relieved by rpoB mutations. J Biol Chem 267:2337–2344
    [Google Scholar]
  25. Williams S. G., Greenwood J. A., Jones C. W. 1994; The effect of nutrient limitation on glycerol uptake and metabolism in continuous cultures of Pseudomonas aeruginosa. Microbiology 140:2961–2969
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-5-1567
Loading
/content/journal/micro/10.1099/00221287-143-5-1567
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error