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. 1969 Feb;97(2):535–543. doi: 10.1128/jb.97.2.535-543.1969

Control of Mixed-Substrate Utilization in Continuous Cultures of Escherichia coli

Richard S Silver a,1, Richard I Mateles a
PMCID: PMC249724  PMID: 4886282

Abstract

The chemostat culture technique was used to study the control mechanisms which operate during utilization of mixtures of glucose and lactose and glucose and l-aspartic acid by populations of Escherichia coli B6. Constitutive mutants were rapidly selected during continuous culture on a mixture of glucose and lactose, and the β-galactosidase level of the culture increased greatly. After mutant selection, the specific β-galactosidase level of the culture was a decreasing function of growth rate. In cultures of both the inducible wild type and the constitutive mutant, glucose and lactose were simultaneously utilized at moderate growth rates, whereas only glucose was used in the inducible cultures at high growth rates. Catabolite repression was shown to be the primary mechanism of control of β-galactosidase level and lactose utilization in continuous culture on mixed substrates. In batch culture, as in the chemostat, catabolite repression acting by itself on the lac enzymes was insufficient to prevent lactose utilization or cause diauxie. Interference with induction of the lac operon, as well as catabolite repression, was necessary to produce diauxic growth. Continuous cultures fed mixtures of glucose and l-aspartic acid utilized both substrates at moderate growth rates, even though the catabolic enzyme aspartase was linearly repressed with increasing growth rate. Although the repression of aspartase paralleled the catabolite repression of β-galactosidase, l-aspartic acid could be utilized even at very low levels of the catabolic enzyme because of direct anabolic incorporation into protein.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ASENSIO C., AVIGAD G., HORECKER B. L. PREFERENTIAL GALACTOSE UTILIZATION IN A MUTANT STRAIN OF E. COLI. Arch Biochem Biophys. 1963 Dec;103:299–309. doi: 10.1016/0003-9861(63)90419-3. [DOI] [PubMed] [Google Scholar]
  2. Adhya S., Echols H. Glucose effect and the galactose enzymes of Escherichia coli: correlation between glucose inhibition of induction and inducer transport. J Bacteriol. 1966 Sep;92(3):601–608. doi: 10.1128/jb.92.3.601-608.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CLARK D. J., MARR A. G. STUDIES ON THE REPRESSION OF BETA-GALACTOSIDASE IN ESCHERICHIA COLI. Biochim Biophys Acta. 1964 Oct 23;92:85–94. doi: 10.1016/0926-6569(64)90272-x. [DOI] [PubMed] [Google Scholar]
  4. COHN M., HORIBATA K. Inhibition by glucose of the induced synthesis of the beta-galactoside-enzyme system of Escherichia coli. Analysis of maintenance. J Bacteriol. 1959 Nov;78:601–612. doi: 10.1128/jb.78.5.601-612.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. COHN M., HORIBATA K. Physiology of the inhibition by glucose of the induced synthesis of the beta-galactosideenzyme system of Escherichia coli. J Bacteriol. 1959 Nov;78:624–635. doi: 10.1128/jb.78.5.624-635.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dobrogosz W. J. Altered end-product patterns and catabolite repression in Escherichia coli. J Bacteriol. 1966 Jun;91(6):2263–2269. doi: 10.1128/jb.91.6.2263-2269.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Epps H. M., Gale E. F. The influence of the presence of glucose during growth on the enzymic activities of Escherichia coli: comparison of the effect with that produced by fermentation acids. Biochem J. 1942 Sep;36(7-9):619–623. doi: 10.1042/bj0360619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HAYASHI S., LIN E. C. CAPTURE OF GLYCEROL BY CELLS OF ESCHERICHIA COLI. Biochim Biophys Acta. 1965 Mar 29;94:479–487. doi: 10.1016/0926-6585(65)90056-7. [DOI] [PubMed] [Google Scholar]
  9. HOLMES R., SHEININ R., CROCKER B. F. A study of the permeability barrier to beta-galactosides in Escherichia coli B. Can J Biochem Physiol. 1961 Jan;39:45–54. doi: 10.1139/o61-006. [DOI] [PubMed] [Google Scholar]
  10. HORIUCHI T., TOMIZAWA J. I., NOVICK A. Isolation and properties of bacteria capable of high rates of beta-galactosidase synthesis. Biochim Biophys Acta. 1962 Jan 22;55:152–163. doi: 10.1016/0006-3002(62)90941-1. [DOI] [PubMed] [Google Scholar]
  11. Hayashi S. I., Lin E. C. Product induction of glycerol kinase in Escherichia coli. J Mol Biol. 1965 Dec;14(2):515–521. doi: 10.1016/s0022-2836(65)80200-5. [DOI] [PubMed] [Google Scholar]
  12. KOCH A. L. The inactivation of the transport mechanism for beta-galactosides of Escherichia coli under various physiological conditions. Ann N Y Acad Sci. 1963 Jan 21;102:602–620. doi: 10.1111/j.1749-6632.1963.tb13663.x. [DOI] [PubMed] [Google Scholar]
  13. KOCH J. P., HAYASHI S., LIN E. C. THE CONTROL OF DISSIMILATION OF GLYCEROL AND L-ALPHA-GLYCEROPHOSPHATE IN ESCHERICHIA COLI. J Biol Chem. 1964 Sep;239:3106–3108. [PubMed] [Google Scholar]
  14. Kamogawa A., Kurahashi K. Inhibitory effect of glucose on the growth of a mutant strain of Escherichia coli defective in glucose transport system. J Biochem. 1967 Feb;61(2):220–230. doi: 10.1093/oxfordjournals.jbchem.a128534. [DOI] [PubMed] [Google Scholar]
  15. Koch A. L. Kinetics of permease catalyzed transport. J Theor Biol. 1967 Feb;14(2):103–130. doi: 10.1016/0022-5193(67)90109-9. [DOI] [PubMed] [Google Scholar]
  16. Kotyk A. Properties of the sugar carrier in baker's yeast. II. Specificity of transport. Folia Microbiol (Praha) 1967;12(2):121–131. doi: 10.1007/BF02896872. [DOI] [PubMed] [Google Scholar]
  17. LEDERBERG J. The beta-d-galactosidase of Escherichia coli, strain K-12. J Bacteriol. 1950 Oct;60(4):381–392. doi: 10.1128/jb.60.4.381-392.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. LOOMIS W. F., Jr, MAGASANIK B. THE RELATION OF CATABOLITE REPRESSION TO THE INDUCTION SYSTEM FOR BETA-GALACTOSIDASE IN ESCHERICHIA COLI. J Mol Biol. 1964 Mar;8:417–426. doi: 10.1016/s0022-2836(64)80205-9. [DOI] [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. Loomis W. F., Jr, Magasanik B. Genetic control of catabolite repression of the lac operon in Escherichia coli. Biochem Biophys Res Commun. 1965 Jul 12;20(2):230–234. doi: 10.1016/0006-291x(65)90351-7. [DOI] [PubMed] [Google Scholar]
  21. Loomis W. F., Jr, Magasanik B. Glucose-lactose diauxie in Escherichia coli. J Bacteriol. 1967 Apr;93(4):1397–1401. doi: 10.1128/jb.93.4.1397-1401.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. MAGASANIK B. Catabolite repression. Cold Spring Harb Symp Quant Biol. 1961;26:249–256. doi: 10.1101/sqb.1961.026.01.031. [DOI] [PubMed] [Google Scholar]
  23. MAGASANIK B., NEIDHARDT F. C. The effect of glucose on the induced biosynthesis of bacterial enzymes in the presence and absence of inducing agents. Biochim Biophys Acta. 1956 Aug;21(2):324–334. doi: 10.1016/0006-3002(56)90016-6. [DOI] [PubMed] [Google Scholar]
  24. MANDELSTAM J. The repression of constitutive beta-galactosidase in Escherichia coli by glucose and other carbon sources. Biochem J. 1962 Mar;82:489–493. doi: 10.1042/bj0820489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. MCFALL E., MANDELSTAM J. SPECIFIC METABOLIC REPRESSION OF THREE INDUCED ENZYMES IN ESCHERICHIA COLI. Biochem J. 1963 Nov;89:391–398. doi: 10.1042/bj0890391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Matsushita S., Iwami N., Nitta Y. Colorimetric estimation of amino acids and peptides with the Folin phenol reagent. Anal Biochem. 1966 Aug;16(2):365–371. doi: 10.1016/0003-2697(66)90168-0. [DOI] [PubMed] [Google Scholar]
  27. Moses V., Prevost C. Catabolite repression of beta-galactosidase synthesis in Escherichia coli. Biochem J. 1966 Aug;100(2):336–353. doi: 10.1042/bj1000336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. NEIDHARDT F. C. Mutant of Aerobacter aerogenes lacking glucose repression. J Bacteriol. 1960 Oct;80:536–543. doi: 10.1128/jb.80.4.536-543.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. NOVICK A., HORIUCHI T. Hyper-production of beta-galactosidase by Escherichia coli bacteria. Cold Spring Harb Symp Quant Biol. 1961;26:239–245. doi: 10.1101/sqb.1961.026.01.029. [DOI] [PubMed] [Google Scholar]
  30. Palmer J., Moses V. Involvement of the lac regulatory genes in catabolite repression in Escherichia coli. Biochem J. 1967 May;103(2):358–366. doi: 10.1042/bj1030358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Prevost C., Moses V. Pool sizes of metabolic intermediates and their relation to glucose repression of beta-galactosidase synthesis in Escherichia coli. Biochem J. 1967 May;103(2):349–357. doi: 10.1042/bj1030349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. REVEL H. R., LURIA S. E., ROTMAN B. Biosynthesis of B-D-galactosidase controlled by phage-carried genes. I. Induced beta-D-galactosidase biosynthesis after transduction of gene z-plus by phage. Proc Natl Acad Sci U S A. 1961 Dec 15;47:1956–1967. doi: 10.1073/pnas.47.12.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zwaig N., Lin E. C. Feedback inhibition of glycerol kinase, a catabolic enzyme in Escherichia coli. Science. 1966 Aug 12;153(3737):755–757. doi: 10.1126/science.153.3737.755. [DOI] [PubMed] [Google Scholar]

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