Antonie van Leeuwenhoek

, Volume 95, Issue 4, pp 305–310 | Cite as

Induction of cadBA in an Escherichia coli lysine auxotroph transformed with a cad-gfp transcriptional fusion

Original Paper
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Abstract

CadBA functions as a part of overall Escherichia coli response to low extracellular pH. A gfpmut3 structural gene transcriptionally fused to the cadBA promoter (Pcad) was used as a reporter to monitor changes in intracellular lysine as a potential factor influencing cadBA induction. Different patterns of cadBA induction were observed in two E. coli strains with different lysine biosynthetic capabilities. In E. coli ZK126 (pJBA25-Pcad), a lysine prototroph, maximum levels of induction were detected 3 h after the transfer of bacterial cells under inducing conditions (pH 5.8; 3.4 μM extracellular lysine). The induction subsequently decreased until hour 7 after which no further change in expression was observed. However, in the lysine depleted strain E. coli ATCC 23812 (pJBA25-Pcad) which is an auxotroph for lysine, no decrease in cadBA expression was observed over time under the same induction conditions. Although no time dependent statistical differences in intracellular lysine were observed, bacterial cells depleted for no longer than 4 h (1.38 ± 0.25 μmol lysine/g cell dry weight) exhibited more rapid induction of cadBA (after 3 h) and a lower maximum level of induction compared to cells with relatively lower intracellular lysine (approximately 1.08 μmol/g cell dry weight). For the latter, the detectable level of induction was delayed for 1 h but the maximum level of induction response was higher.

Keywords

Escherichia coli Lysine auxotroph cad operon Green fluorescent protein Transcriptional fusion Intracellular lysine 

References

  1. Adelberg EA, Mandel M, Chein CCG (1965) Optimal conditions for mutagenesis by N-methyl-N′-nitro-N-nitrosoguanidine in Escherichia coli K12. Biochem Biophys Res Commun 18:788–795. doi:10.1016/0006-291X(65)90855-7 CrossRefGoogle Scholar
  2. Andersen JB, Sternberg C, Poulsen LK, Bjørn SP, Givskov M, Molin S (1998) New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Appl Environ Microbiol 64:2240–2246PubMedGoogle Scholar
  3. Auger EA, Redding KE, Plumb T, Childs LC, Meng S-Y, Bennett GN (1989) Construction of lac fusions to the inducible arginine- and lysine decarboxylase genes of Escherichia coli K12. Mol Microbiol 3:609–620. doi:10.1111/j.1365-2958.1989.tb00208.x PubMedCrossRefGoogle Scholar
  4. Bodini S, Nunziangeli L, Santori F (2007) Influence of amino acids on low-density Escherichia coli responses to nutrient downshifts. J Bacteriol 189:3099–3105. doi:10.1128/JB.01753-06 PubMedCrossRefGoogle Scholar
  5. Chalova VI, Woodward CL, Ricke SC (2008) A cad-gfpmut3 plasmid construct in Escherichia coli for gene induction-based quantification of lysine in acid hydrolysates of feedstuffs. Lett Appl Microbiol 46:107–112PubMedGoogle Scholar
  6. Connell N, Han Z, Moreno F, Kolter R (1987) An E coli promoter induced by the cessation of growth. Mol Microbiol 1:195–201. doi:10.1111/j.1365-2958.1987.tb00512.x PubMedCrossRefGoogle Scholar
  7. Gale EF (1946) The bacterial amino acid decarboxylases. Adv Enzymol 6:1–32Google Scholar
  8. Heyde M, Portalier R (1990) Acid shock proteins of Escherichia coli. FEMS Microbiol Lett 69:19–26. doi:10.1111/j.1574-6968.1990.tb04168.x CrossRefGoogle Scholar
  9. Hickey EW, Hirshfield IN (1990) Low-pH-induced effects on patterns of protein synthesis and on internal pH in Escherichia coli and Salmonella typhimurium. Appl Environ Microbiol 56:1038–1045PubMedGoogle Scholar
  10. Ingraham JL, Marr AG (1996) Effect of temperature, pressure, pH, and osmotic stress on growth. In: Neidhardt FC, Curtiss RIII, Ingraham JL, Lin ECC, Low KB, Magasanik B, Reznikoff WS, Riley M, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella: Cellular and molecular biology, vol 2, 2nd edn. ASM Press, Washington, D.C, pp 1570–1578Google Scholar
  11. Kiefer P, Heinzle E, Zelder O, Wittmann C (2004) Comparative metabolic flux analysis of lysine-producing Corynebacterium glutamicum cultured on glucose or fructose. Appl Environ Microbiol 70:229–239. doi:10.1128/AEM.70.1.229-239.2004 PubMedCrossRefGoogle Scholar
  12. Kostrzynska M, Leung KT, Lee H, Trevors JT (2002) Green fluorescent protein-based biosensor for detecting SOS-inducing activity of genotoxic compounds. J Microbiol Methods 48:43–51. doi:10.1016/S0167-7012(01)00335-9 PubMedCrossRefGoogle Scholar
  13. Krömer JO, Sorgenfrei O, Klopprogge K, Heinzle E, Wittmann C (2004) In-depth profiling of lysine-producing Corynebacterium glutamicum by combined analysis of the transcriptome, metabolome, and fluxome. J Bacteriol 186:1769–1784. doi:10.1128/JB.186.6.1769-1784.2004 PubMedCrossRefGoogle Scholar
  14. Li X, Erickson AM, Ricke SC (1999) Comparison of minimal media and inoculum concentration to decrease the lysine growth assay response time of an Escherichia coli lysine auxotroph mutant. J Rapid Methods Autom Microbiol 7:279–290Google Scholar
  15. Meng S-Y, Bennett GN (1992a) Regulation of the Escherichia coli cad operon: location of a site required for acid induction. J Bacteriol 174:2670–2678PubMedGoogle Scholar
  16. Meng S-Y, Bennett GN (1992b) Nucleotide sequence of the Escherichia coli cad operon: a system for neutralization of low extracellular pH. J Bacteriol 174:2659–2669PubMedGoogle Scholar
  17. Møller S, Sternberg C, Andersen JB, Christensen BB, Ramos JL, Givskov M, Molin S (1998) In situ expression in mixed-culture biofilms: evidence of metabolic interactions between community members. Appl Environ Microbiol 64:721–732PubMedGoogle Scholar
  18. Neely MN, Olson ER (1996) Kinetics of expression of the Escherichia coli cad operon as a function of pH and lysine. J Bacteriol 178:5522–5528PubMedGoogle Scholar
  19. Neely MN, Dell CL, Olson ER (1994) Roles of LysP and CadC in mediating the lysine requirement for acid induction of the Escherichia coli cad operon. J Bacteriol 176:3278–3285PubMedGoogle Scholar
  20. Raja N, Goodson M, Chui WC, Smith DG, Rowbury RJ (1991) Habituation to acid in Escherichia coli: conditions for habituation and its effect on plasmid transfer. J Appl Bacteriol 70:59–65PubMedGoogle Scholar
  21. Recsei PA, Snell EE (1972) Histidine decarboxylaseless mutants of Lactobacillus 30a: isolation and growth properties. J Bacteriol 112:624–626PubMedGoogle Scholar
  22. Sabo DL, Boeker EA, Byers B, Waron H, Fischer EH (1974) Purification and physical properties of inducible Escherichia coli lysine decarboxylase. Biochem 13:662–670. doi:10.1021/bi00701a005 CrossRefGoogle Scholar
  23. Sambrook J, Frish EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.YGoogle Scholar
  24. Slonczewski JL, Foster JW (1996) pH-regulated genes and survival at extreme pH. In: Neidhardt FC, Curtiss RIII, Ingraham JL, Lin ECC, Low KB, Magasanik B, Reznikoff WS, Riley M, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella: Cellular and molecular biology, vol 1, 2nd edn. ASM Press, Washington, D.C, pp 1539–1549Google Scholar
  25. Slonczewski JL, Rosen BP, Alger JR, Macnab RM (1981) pH homeostasis in Escherichia coli: measurement by 31P nuclear magnetic resonance of methylphosphonate and phosphate. Proc Natl Acad Sci USA 78:6271–6275. doi:10.1073/pnas.78.10.6271 PubMedCrossRefGoogle Scholar
  26. Soksawatmaekhin W, Kuraishi A, Sakata K, Kashiwagi K, Igarshi K (2004) Excretion and uptake of cadaverine by CadB and its physiological functions in Escherichia coli. Mol Microbiol 51:1401–1412. doi:10.1046/j.1365-2958.2003.03913.x PubMedCrossRefGoogle Scholar
  27. Wittmann C, Krömer JO, Kiefer P, Binz T, Heinzle E (2004) Impact of the cold shock phenomenon on quantification of intracellular metabolites in bacteria. Anal Biochem 327:135–139. doi:10.1016/j.ab.2004.01.002 PubMedCrossRefGoogle Scholar
  28. Zilberstein D, Agmon V, Schuldiner S, Padan E (1984) Escherichia coli intracellular pH, membrane potential, and cell growth. J Bacteriol 158:246–252PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • V. I. Chalova
    • 1
    • 2
  • C. L. Woodward
    • 1
  • S. C. Ricke
    • 1
    • 2
    • 3
  1. 1.Department of Poultry ScienceTexas A&M UniversityCollege StationUSA
  2. 2.Center for Food Safety, Department of Food Science, and Department of Poultry ScienceUniversity of ArkansasFayettevilleUSA
  3. 3.Department of Food ScienceUniversity of ArkansasFayettevilleUSA

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