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Fisiologi E Coli Dimedium LB
Fisiologi E Coli Dimedium LB
23
0021-9193/07/$08.00⫹0 doi:10.1128/JB.01368-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.
Luria-Bertani broth supports Escherichia coli growth to an optical density at 600 nm (OD600) of 7. Surpris-
ingly, however, steady-state growth ceases at an OD600 of 0.3, when the growth rate slows down and cell mass
decreases. Growth stops for lack of a utilizable carbon source. The carbon sources for E. coli in Luria-Bertani
broth are catabolizable amino acids, not sugars.
The widely used rich medium called Luria-Bertani broth is undertook a closer study of the physiology of E. coli K-12 in
8746
VOL. 189, 2007 NOTES 8747
TABLE 1. Composition of Luria-Bertani broth It should be noted that the composition of Luria-Bertani
Concn (mM)
broth is not constant. Variations occur with age (L-tryptophan,
Metabolite(s)
Allele conferring determined by: for example, is degraded, especially in the light), with time of
auxotrophya autoclaving (affecting the degree of deamidation of L-aspara-
E. colib HPLCc
gine and L-glutamine), and from batch to batch. These varia-
L-Alanine NDd 5.8 tions may account for those cases in which the bioassay gave a
L-Arginine argA::Tn10 2.0 2.8
higher estimate than HPLC analysis.
L-Asparagine asnA31 asnB50::Tn5 ⱕ2.1e
L-Aspartate NDd Catabolyzable amino acids are clearly plentiful in Luria-
L-Asp ⫹ L-Asn 7.3 Bertani broth, presumably in the form of oligopeptides. It has
L-Cysteine cysG 1.7 0.4 been reported that wild-type E. coli K-12 growing in tryptone
L-Glutamic acid ⌬(gltA-sucB)::Spcr 11.2 broth sequentially catabolizes L-serine, L-aspartate, L-trypto-
L-Glutamine glnA1857 0.6
L-Glu ⫹ L-Gln 19.1 phan, L-glutamate, glycine, L-threonine, and L-alanine (9). Se-
Glycine glyA42::Tn5 18 4.0 quential catabolism probably occurs in Luria-Bertani broth as
L-Histidine hisG213::Tn10 0.75 1.4 well, since a diauxic lag was observed (12). It may follow the
L-Isoleucine ilvA454 2.3 5.4 same order, although one would also expect L-proline, a good
L-Leucine leu-82::Tn10 7.9 8.8
acid equivalents. These are probably depleted sequentially subunit of RNA polymerase in Escherichia coli is controlled at the levels of
transcription, translation, and protein stability. Genes Dev. 8:1600–1612.
during the postexponential phase of growth. During this pe- 6. McFall, E., and E. B. Newman. 1996. Amino acids as carbon sources, p.
riod, the physiological state of the cells varies constantly. Fur- 358–379. In F. C. Neidhardt, R. Curtiss III, J. L. Ingraham, E. C. C. Lin, K. B.
thermore, even using the same strain at the same OD600, if the Low, B. Magasanik, W. S. Reznikoff, M. Riley, M. Schaechter, and H. E.
Umbarger (ed.), Escherichia coli and Salmonella: cellular and molecular
latter is above 0.3 the cells’ physiological state will not neces- biology. ASM Press, Washington, D.C.
sarily be reproducible in different experiments since the com- 7. Miller, C. G. 1996. Protein degradation and proteolytic modification, p.
position of Luria-Bertani broth can vary. Caveat experimenta- 938–954. In F. C. Neidhardt, R. Curtiss III, J. L. Ingraham, E. C. C. Lin, K. B.
Low, B. Magasanik, W. S. Reznikoff, M. Riley, M. Schaechter, and H. E.
tor! Umbarger (ed.), Escherichia coli and Salmonella: cellular and molecular
biology. ASM Press, Washington, D.C.
8. Neidhardt, F. C., J. L. Ingraham, and M. Schaechter. 1990. Physiology of the
We thank Conrad Woldringh for helpful discussion.
bacterial cell. Sinauer Associates, Sunderland, MA.
9. Prüss, B. M., J. M. Nelms, C. Park, and A. J. Wolfe. 1994. Mutations in
REFERENCES NADH:ubiquinone oxidoreductase of Escherichia coli affect growth on
mixed amino acids. J. Bacteriol. 176:2143–2150.
1. Bachmann, B. J. 1996. Derivations and genotypes of some mutant derivatives of 10. Riley, M., T. Abe, M. B. Arnaud, M. K. B. Berlyn, F. R. Blattner, R. R.
Escherichia coli K-12, p. 2460–2488. In F. C. Neidhardt, R. Curtiss III, J. L. Chaudhuri, J. D. Glasner, T. Horiuchi, I. M. Keseler, T. Kosuge, H. Mori,
Ingraham, E. C. C. Lin, K. B. Low, B. Magasanik, W. S. Reznikoff, M. Riley, M. N. T. Perna, G. Plunkett III, K. E. Rudd, M. H. Serres, G. H. Thomas, N. R.
Schaechter, and H. E. Umbarger (ed.), Escherichia coli and Salmonella: cellular Thomson, D. Wishart, and B. L. Wanner. 2006. Escherichia coli K-12: a