Advertisement

Amino Acids

, Volume 39, Issue 3, pp 727–737 | Cite as

Glutamate-induced metabolic changes in Lactococcus lactis NCDO 2118 during GABA production: combined transcriptomic and proteomic analysis

  • Roberto Mazzoli
  • Enrica PessioneEmail author
  • Magali Dufour
  • Valérie Laroute
  • Maria Gabriella Giuffrida
  • Carlo Giunta
  • Muriel Cocaign-Bousquet
  • Pascal Loubière
Original Article

Abstract

GABA is a molecule of increasing nutraceutical interest due to its modulatory activity on the central nervous system and smooth muscle relaxation. Potentially probiotic bacteria can produce it by glutamate decarboxylation, but nothing is known about the physiological modifications occurring at the microbial level during GABA production. In the present investigation, a GABA-producing Lactococcus lactis strain grown in a medium supplemented with or without glutamate was studied using a combined transcriptome/proteome analysis. A tenfold increase in GABA production in the glutamate medium was observed only during the stationary phase and at low pH. About 30 genes and/or proteins were shown to be differentially expressed in glutamate-stimulated conditions as compared to control conditions, and the modulation exerted by glutamate on entire metabolic pathways was highlighted by the complementary nature of transcriptomics and proteomics. Most glutamate-induced responses consisted in under-expression of metabolic pathways, with the exception of glycolysis where either over- or under-expression of specific genes was observed. The energy-producing arginine deiminase pathway, the ATPase, and also some stress proteins were down-regulated, suggesting that glutamate is not only an alternative means to get energy, but also a protective agent against stress for the strain studied.

Keywords

GABA Glutamate decarboxylase Branched chain amino acids ADI route Stress ATPase 

Notes

Acknowledgments

This research, conceived in the context of the CESQTA (Center for Food Safety and Quality, Piedmont, Italy), has been supported by GALILEE-EGIDE project. This paper is published despite the effects of the Italian law 133/08. This law drastically reduces public funds to public Italian universities, which is particularly dangerous for scientific free research, and it will prevent young researchers from getting a position, either temporary or tenured, in Italy.

Supplementary material

726_2010_507_MOESM1_ESM.doc (82 kb)
Supplementary material 1 (DOC 82 kb)

References

  1. Arsène-Ploetze F, Kugler V, Martinussen J, Bringel F (2006) Expression of the pyr operon of Lactobacillus plantarum is regulated by inorganic carbon availability through a second regulator, PyrR2, homologous to the pyrimidine-dependent regulator PyrR1. J Bacteriol 188:8607–8616CrossRefPubMedGoogle Scholar
  2. Budin-Verneuil A, Maguin E, Auffray Y, Dusko Ehrlich S, Pichereau V (2004) An essential role for arginine catabolism in the acid tolerance of Lactococcus lactis MG1363. Lait 84:61–68CrossRefGoogle Scholar
  3. Cox B, Kislinger T, Emili A (2005) Integrating gene and protein expression data: pattern analysis and profile mining. Methods 35:303–314CrossRefPubMedGoogle Scholar
  4. Exterkate FA, de Veer GJCM (1987) Purification and some properties of a membrane-bound aminopeptidase A from Streptococcus cremoris. Appl Environ Microbiol 53:577–583PubMedGoogle Scholar
  5. Fernández M, Zúñiga M (2006) Amino acid catabolic pathways of lactic acid bacteria. Crit Rev Microbiol 32:155–183CrossRefPubMedGoogle Scholar
  6. Frees D, Savijoki K, Varmanen P, Ingmer H (2007) Clp ATPases and ClpP proteolytic complexes regulate vital biological processes in low GC, Gram-positive bacteria. Mol Microbiol 63:1285–1295CrossRefPubMedGoogle Scholar
  7. Giuffrida MG, Pessione E, Mazzoli R, Della Valle G et al (2001) Media containing aromatic compounds induce peculiar proteins in Acinetobacter radioresistens, as revealed by proteome analysis. Electrophoresis 22:1705–1711CrossRefPubMedGoogle Scholar
  8. Higuchi T, Hayashi H, Abe K (1997) Exchange of glutamate and gamma-aminobutyrate in a Lactobacillus strain. J Bacteriol 179:3362–3364PubMedGoogle Scholar
  9. Hughes MJ, Moore JC, Lane JD, Wilson R et al (2002) Identification of major outer surface proteins of Streptococcus agalactiae. Infect Immun 70:1254–1259CrossRefPubMedGoogle Scholar
  10. Inoue K, Shirai T, Ochiai H, Kasao M et al (2003) Blood-pressure-lowering effect of a novel fermented milk containing gamma-aminobutyric acid (GABA) in mild hypertensives. Eur J Clin Nutr 57:490–495CrossRefPubMedGoogle Scholar
  11. Jorgensen CM, Hammer K, Jensen PR, Martinussen J (2004) Expression of the pyrG gene determines the pool sizes of CTP and dCTP in Lactococcus lactis. Eur J Biochem 271:2438–2445CrossRefPubMedGoogle Scholar
  12. Kim D, San BH, Moh SH, Park H, Kim DY, Lee S, Kim KK (2009) Structural basis for the substrate specificity of PepA from Streptococcus pneumoniae, a dodecameric tetrahedral protease. Biochem Biophys Res Commun (Epub ahead of print)Google Scholar
  13. Konings WN (2002) The cell membrane and the struggle for life of lactic acid bacteria. Antonie Van Leeuwenhoek 82:3–27CrossRefPubMedGoogle Scholar
  14. Konings WN (2006) Microbial transport: adaptations to natural environments. Antonie van Leeuwenhoek 90:325–342CrossRefPubMedGoogle Scholar
  15. Lucas P, Landete J, Coton M, Coton E, Lonvaud-Funel A (2003) The tyrosine decarboxylase operon of Lactobacillus brevis IOEB 9809: characterization and conservation in tyramine-producing bacteria. FEMS Microbiol Lett 229:65–71CrossRefPubMedGoogle Scholar
  16. Lucas PM, Wolken WAM, Claisse O, Lolkema JS, Lonvaud-Funel A (2005) Histamine-producing pathway encoded on an unstable plasmid in Lactobacillus hilgardiii 0006. Appl Environ Microbiol 71:1417–1424CrossRefPubMedGoogle Scholar
  17. Maligoy M, Mercade M, Cocaign-Bousquet M, Loubière P (2008) Transcriptome analysis of Lactococcus lactis in coculture with Saccharomyces cerevisiae. Appl Environ Microbiol 74:485–494CrossRefPubMedGoogle Scholar
  18. Martos GI, Minahk CJ, de Valdez GF, Morero R (2007) Effects of protective agents on membrane fluidity of freeze-dried Lactobacillus delbrueckii ssp. bulgaricus. Lett Appl Microbiol 45:282–288CrossRefPubMedGoogle Scholar
  19. Millichap JG, Yee MM (2003) The diet factor in pediatric and adolescent migraine. Pediatr Neurol 28:9–15CrossRefPubMedGoogle Scholar
  20. Neuhoff V, Arold N, Taube D, Ehrhardt W (1988) Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250. Electrophoresis 9:255–262CrossRefPubMedGoogle Scholar
  21. Nicoloff H, Elagoz A, Arsène-Ploetze F, Kammerer B et al (2005) Repression of the pyr operon in Lactobacillus plantarum prevents ist ability to grow at low carbon dioxide levels. J Bacteriol 187:2093–2104CrossRefPubMedGoogle Scholar
  22. Novak L, Cocaign-Bousquet M, Lindley ND, Loubière P (1997) Metabolism and energetics of Lactococcus lactis during growth in complex or synthetic media. Appl Environ Microbiol 63:2665–2670PubMedGoogle Scholar
  23. Pessione E, Mazzoli R, Giuffrida MG, Lamberti C et al (2005) A proteomic approach to studying biogenic amine producing lactic acid bacteria. Proteomics 5:687–698CrossRefPubMedGoogle Scholar
  24. Pessione E, Pessione A, Lamberti C, Coïsson JD et al (2009) First evidence of a membrane-bound, tyramine and beta-phenylethylamine producing, tyrosine decarboxylase in Enterococcus faecalis: a two-dimensional electrophoresis proteomic study. Proteomics 9:2695–2710CrossRefPubMedGoogle Scholar
  25. Poolman B, Konings WN (1988) Relation of growth of Streptococcus lactis and Streptococcus cremoris to amino acid transport. J Bacteriol 170:700–707PubMedGoogle Scholar
  26. Rabilloud T (1998) Use of thiourea to increase the solubility of membrane proteins in two-dimensional electrophoresis. Electrophoresis 19:758–760CrossRefPubMedGoogle Scholar
  27. Raikar LS, Vallejo J, Lloyd PG, Hardin CD (2006) Overexpression of caveolin-1 results in increased plasma membrane targeting of glycolytic enzymes: the structural basis for a membrane associated metabolic compartment. J Cell Biochem 98:861–871CrossRefPubMedGoogle Scholar
  28. Redon E, Loubière P, Cocaign-Bousquet M (2005) Role of mRNA stability during genome-wide adaptation of Lactococcus lactis to carbon starvation. J Biol Chem 280:36380–36385CrossRefPubMedGoogle Scholar
  29. Sanders JW, Leenhouts K, Burghoorn J, Brands JR et al (1998) A chloride-inducible acid resistance mechanism in Lactococcus lactis and its regulation. Mol Microbiol 27:299–310CrossRefPubMedGoogle Scholar
  30. Schelp E, Worley S, Monzingo AF, Ernst S, Robertus JD (2001) pH-induced structural changes regulate histidine decarboxylase activity in Lactobacillus 30a. J Mol Biol 306:727–732CrossRefPubMedGoogle Scholar
  31. Singh OV, Nagaraj NS (2006) Transcriptomics, proteomics and interactomics: unique approaches to track the insights of bioremediation. Brief Funct Genomic Proteomic 4:355–362CrossRefPubMedGoogle Scholar
  32. Siragusa S, De Angelis M, Di Cagno R, Rizzello CG, Coda R, Gobbetti M (2007) Synthesis of gamma-aminobutyric acid by lactic acid bacteria isolated from a variety of Italian cheeses. Appl Environ Microbiol 73:7283–7290CrossRefPubMedGoogle Scholar
  33. Thompson (1987) Sugar transport in the lactic acid bacteria. In: Reizer J, Peterkofsky A (eds) Sugar transport and metabolism in gram-positive bacteria. Ellis Horwood, Chichester, pp 15–38Google Scholar
  34. Tramonti A, De Canio M, Delany I, Scarlato V, De Biase D (2006) Mechanisms of transcription activation exerted by GadX and GadW at the gadA and gadBC gene promoters of the glutamate-based acid resistance system in Escherichia coli. J Bacteriol 188:8118–8127CrossRefPubMedGoogle Scholar
  35. Van de Guchte M, Serror P, Chervaux C, Smokvina T et al (2002) Stress responses in lactic acid bacteria. Antonie van Leeuwenhoek 82:187–216CrossRefPubMedGoogle Scholar
  36. Vercauteren FG, Arckens L, Quirion R (2007) Applications and current challenges of proteomic approaches, focusing on two-dimensional electrophoresis. Amino Acids 33:405–414CrossRefPubMedGoogle Scholar
  37. Wilkins JC, Beighton D, Homer KA (2003) Effect of acidic pH on expression of surface-associated proteins of Streptococcus oralis. Appl Environ Microbiol 69:5290–5296CrossRefPubMedGoogle Scholar
  38. Willemoës M, Kilstrup M, Roepstorff P, Hammer K (2002) Proteome analysis of a Lactococcus lactis strain overexpressing gapA suggests that the gene product is an auxiliary glyceraldehyde 3-phosphate dehydrogenase. Proteomics 2:1041–1046CrossRefPubMedGoogle Scholar
  39. Yan D (2007) Protection of the glutamate pool concentration in enteric bacteria. Proc Natl Acad Sci USA 104:9475–9480CrossRefPubMedGoogle Scholar
  40. Zuobi-Hasona K, Crowley PJ, Hasona A, Bleiweis AS, Brady LJ (2005) Solubilization of cellular membrane proteins from Streptococcus mutans for two-dimensional gel electrophoresis. Electrophoresis 26:1200–1205CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Roberto Mazzoli
    • 1
    • 2
    • 4
    • 5
  • Enrica Pessione
    • 1
    Email author
  • Magali Dufour
    • 1
  • Valérie Laroute
    • 2
    • 4
    • 5
  • Maria Gabriella Giuffrida
    • 3
  • Carlo Giunta
    • 1
  • Muriel Cocaign-Bousquet
    • 2
    • 4
    • 5
  • Pascal Loubière
    • 2
    • 4
    • 5
  1. 1.Dipartimento di Biologia Animale e dell’UomoUniversità di TorinoTorinoItaly
  2. 2.Université de Toulouse; INSA,UPS,INP; LISBPToulouseFrance
  3. 3.ISPA-CNR. c/o Bioindustry Park CanaveseColleretto GiacosaItaly
  4. 4.INRAUMR792 Ingénierie des Systèmes Biologiques et des ProcédésToulouseFrance
  5. 5.CNRSUMR5504ToulouseFrance

Personalised recommendations