Skip to main content
SpringerLink
Log in

Chlorobenzoate inhibits growth and induces stress proteins in the PCB-degrading bacterium Burkholderia xenovorans LB400

Archives of Microbiology volume 188, pages 289–297 (2007)Cite this article

Abstract

Aerobic bacteria, such as Burkholderia xenovorans LB400, are able to degrade a wide range of polychlorobiphenyls (PCBs). Generally, these bacteria are not able to transform chlorobenzoates (CBAs), which accumulate during PCB degradation. In this study, the effects of CBAs on the growth, the morphology and the proteome of Burkholderia xenovorans LB400 were analysed. 4-CBA and 2-CBA were observed to inhibit the growth of strain LB400 on glucose. Strain LB400 exposed to 4-CBA exhibited increased number and size of electron-dense granules in the cytoplasm, which could be polyphosphates. Two-dimensional (2-D) polyacrylamide gel electrophoresis was used to characterise the molecular response of strain LB400 to 4-CBA. This compound induced the enzymes BenD and CatA of benzoate and catechol catabolic pathways. The induction of molecular chaperones DnaK and HtpG by 4-CBA indicated that the exposure to this compound constitutes a stressful condition for this bacterium. Additionally, the induction of some Krebs cycle enzymes was observed, probably as response to cellular energy requirements. This study contributes to the knowledge on the effects of CBA on the PCB-degrader Burkholderia xenovorans LB400.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Abbreviations

CBAs:

Chlorobenzoates

PCBs:

Polychlorobiphenyls

2-D:

Two-dimensional

DNP:

2,4-Dinitrophenol

2,4-D:

2,4-Dichlorophenoxyacetic acid

CFU:

Colony-forming units

References

  • Agulló L, Cámara B, Martínez P, Latorre V, Seeger M (2007) Response to (chloro)biphenyls of the polychlorobiphenyl-degrader Burkholderia xenovorans LB400 involves stress proteins also induced by heat shock and oxidative stress. FEMS Microbiol Lett 267:167–175

    Article  PubMed  CAS  Google Scholar 

  • Barriault D, Lépine F, Mohammadi M, Milot S, Leberre N, Sylvestre M (2004) Revising the regiospecificity of Burkholderia xenovorans biphenyl dioxygenase toward 2,2′-dichlorobiphenyl and 2,3,2′,3′-tetrachlorobiphenyl. J Biol Chem 279:47489–47496

    Article  PubMed  CAS  Google Scholar 

  • Bébien M, Lagniel G, Garin J, Touati D, Vermeglio A, Labarre J (2002) Involvement of superoxide dismutases in the response of Escherichia coli to selenium oxides. J Bacteriol 184:1556–1564

    Article  PubMed  CAS  Google Scholar 

  • Bedard DL (1990) Bacterial transformation of polychlorinated biphenyls. In: Kamely D, Chakrabarty A, Omenn GS (eds) Biotechnology and biodegradation. Portfolio Publishing Co., The Woodlands, Texas, pp 369–388

    Google Scholar 

  • Blasco R, Wittich RM, Mallavarapu M, Timmis KN, Pieper DH (1995) From xenobiotic to antibiotic, formation of protoanemonin from 4-chlorocatechol by enzymes of the 3-oxoadipate pathway. J Biol Chem 270:29229–29235

    Article  PubMed  CAS  Google Scholar 

  • Caldas T, Laalami S, Richarme G (2000) Chaperone properties of bacterial elongation factor EF-G and initiation factor IF2. J Biol Chem 275:855–860

    Article  PubMed  CAS  Google Scholar 

  • Cámara B, Herrera C, González M, Couve E, Hofer B, Seeger M (2004) From PCBs to highly toxic metabolites by the biphenyl pathway. Environ Microbiol 6:842–850

    Article  PubMed  CAS  Google Scholar 

  • Cases I, De Lorenzo V (2001) The black cat/white cat principle of signal integration in bacterial promoters. EMBO J 20:1–11

    Article  PubMed  CAS  Google Scholar 

  • Chain P, Denef V, Konstantinidis K, Vergez L, Agulló L, Latorre Reyes V, Hauser L, Córdova M, Gómez L, González M, Land M, Lao V, Larimer F, LiPuma J, Mahenthiralingam E, Malfatti S, Marx C, Parnell J, Ramette A, Richardson P, Seeger M, Smith D, Spilker T, Sul W-J, Tsoi T, Ulrico L, Zhulin I, Tiedje J (2006). Burkholderia xenovorans LB400 harbors a multi-replicon, 9.7 M bp genome shaped for versatility. Proc Natl Acad Sci USA 113:15280–15287

    Article  Google Scholar 

  • Cho YS, Park SH, Kim CK, Oh KH (2000) Induction stress shock proteins DnaK and GroEL by phenoxyherbicide 2,4-D in Burkholderia sp. YK-2 isolated from rice field. Curr Microbiol 41:33–38

    Article  PubMed  CAS  Google Scholar 

  • Denef VJ, Park J, Tsoi TV, Rouillard JM, Zhang H, Wibbenmeyyer JA, Verstraete W, Gulari E, Hashsham SA, Tiedje JM (2004) Biphenyl and benzoate metabolism in a genomic context: outlining genome-wide metabolic networks in Burkholderia xenovorans LB400. Appl Environ Microbiol 70:4961–4970

    Article  PubMed  CAS  Google Scholar 

  • Denef VJ, Patrauchan MA, Florizone C, Park J, Tsoi TV, Verstraete W, Tiedje JM, Eltis LD (2005) Growth substrate- and phase-specific expression of biphenyl, benzoate and C1 metabolic pathways in Burkholderia xenovorans LB400. J Bacteriol 187:7996–8005

    Article  PubMed  CAS  Google Scholar 

  • Duché O, Trémoulet F, Glaser P, Labadie J (2002) Salt stress proteins induced in Listeria monocytogenes. Appl Environ Microbiol 68:1491–1498

    Article  PubMed  CAS  Google Scholar 

  • Gage DJ, Neidhardt FC (1993) Adaptation of Escherichia coli to the uncoupler of oxidative phosphorylation 2,4-dinitrophenol. J Bacteriol 175:7105–7108

    PubMed  CAS  Google Scholar 

  • Haddock JD, Horton JR, Gibson DT (1995) Dihydroxylation and dechlorination of chlorinated biphenyls by purified biphenyl-2,3-dioxygenase from Pseudomonas sp. strain LB400. J Bacteriol 177:20–26

    PubMed  CAS  Google Scholar 

  • Heim S, Lleo M, Bonato B, Guzman CA, Canepari P (2002) The viable but nonculturable state and starvation are different stress responses of Enterococcus faecalis, as determined by proteome analysis. J Bacteriol 184:6739–6745

    Article  PubMed  CAS  Google Scholar 

  • Heipieper HJ, de Bont JAM (1994) Adaptation of Pseudomonas putida S12 to ethanol and toluene at the level of fatty acid composition of membranes. Appl Environ Microbiol 60:4440–4444

    PubMed  CAS  Google Scholar 

  • Kabelitz N, Santos PM, Heipieper HJ (2003) Effect of aliphatic alcohols on growth and degree of saturation of membrane lipids in Acinetobacter calcoaceticus. FEMS Microbiol Lett 220:223–227

    Article  PubMed  CAS  Google Scholar 

  • Kaschabek SR, Kasberg T, Müller D, Mars AE, Janssen DB, Reineke W (1998) Degradation of chloroaromatics: purification and characterization of a novel type of chlorocatechol 2,3-dioxygenase of Pseudomonas putida GJ31. J Bacteriol 180:296–302

    PubMed  CAS  Google Scholar 

  • Kim S-II, Seung-Youl S, Kyung-Wook K, Eun-Mi H, Kye-Heon O (2003) Proteomic analysis of the benzoate degradation pathway in Acinetobacter sp. KS-1. Res Microbiol 154:697–703

    Article  PubMed  CAS  Google Scholar 

  • Krayl M, Benndorf D, Loffhagen N, Babel W (2003) Use of proteomics and physiological characteristics to elucidate ecotoxic effects of methyl tert-butyl ether in Pseudomonas putida KT2440. Proteomics 3:1544–1552

    Article  PubMed  CAS  Google Scholar 

  • Kulaev I, Kulakovskaya T (2000) Polyphosphate and phosphate pump. Annu Rev Microbiol 54:709–734

    Article  PubMed  CAS  Google Scholar 

  • Lambert LA, Abshire K, Blankenhorn D, Slonczewski J (1997) Proteins induced in Escherichia coli by benzoic acid. J Bacteriol 179:7595–7599

    PubMed  CAS  Google Scholar 

  • Leichert LI, Scharf C, Hecker M (2003) Global characterization of disulfide stress in Bacillus subtilis. J Bacteriol 185:1967–1975

    Article  PubMed  CAS  Google Scholar 

  • Loffhagen N, Hartig C, Babel W (2003) Energization of Comamonas testosteroni ATCC 17454 for indicating toxic effects of chlorophenoxy herbicides. Arch Environ Contam Toxicol 45:317–323

    Article  PubMed  CAS  Google Scholar 

  • Lupi CG, Colangelo T, Mason CA (1995) Two-dimensional gel electrophoresis analysis of the response of Pseudomonas putida KT244 to 2-chlorophenol. Appl Environ Microbiol 61:2863–2872

    PubMed  CAS  Google Scholar 

  • Neidle EL, Ornston LN (1987) Benzoate and muconate, structurally dissimilar metabolites, induce expression of catA in Acinetobacter calcoaceticus. J Bacteriol 169:414–415

    PubMed  CAS  Google Scholar 

  • Neidle EL, Shapiro MK, Ornston LN (1987) Cloning and expression in Escherichia coli of Acinetobacter calcoaceticus genes for benzoate degradation. J Bacteriol 169:5496–5503

    PubMed  CAS  Google Scholar 

  • Pieper DH (2005) Aerobic degradation of polychlorinated biphenyls. Appl Microbiol Biotechnol 67:170–191

    Article  PubMed  CAS  Google Scholar 

  • Santos PM, Benndorf D, Sa-Correia I (2004) Insights into Pseudomonas putida KT2440 response to phenol-induce stress by quantitative proteomics. Proteomics 4:2640–2652

    Article  PubMed  CAS  Google Scholar 

  • Seeger M, Timmis K, Hofer B (1995) Conversion of chlorobiphenyls into phenylhexadienoates and benzoates by the enzymes of the upper pathway for polychlorobiphenyl degradation encoded by the bph locus of Pseudomonas sp. strain LB400. Appl Environ Microbiol 61:2654–2658

    PubMed  CAS  Google Scholar 

  • Seeger M, Osorio G, Jerez C (1996) Phosphorylation of GroEL, DnaK and other proteins from Thiobacillus ferrooxidans grown under different conditions. FEMS Microbiol Lett 138:129–134

    Article  PubMed  CAS  Google Scholar 

  • Seeger M, Timmis KN, Hofer B (1997) Bacterial pathways for the degradation of polychlorinated biphenyl. Mar Chem 58:327–333

    Article  CAS  Google Scholar 

  • Seeger M, Zielinski M, Timmis K, Hofer B (1999) Regioespecificity of dioxygenation of di- to pentachlorobiphenyls and their degradation to chlorobenzoates by the bph-encoded catabolic pathway of Burkholderia sp. strain LB400. Appl Environ Microbiol 65:3614–3621

    PubMed  CAS  Google Scholar 

  • Seeger M, Cámara B, Hofer B (2001) Dehalogenation, denitration, dehydroxylation and angular attack of substituted biphenyls and related compounds by a biphenyl dioxygenase. J Bacteriol 183:3548–3555

    Article  PubMed  CAS  Google Scholar 

  • Seeger M, González M, Cámara B, Muñoz L, Ponce E, Mejías L, Mascayano C, Vásquez Y, Sepúlveda-Boza S (2003) Biotransformation of natural and synthetic isoflavonoids by two recombinant microbial enzymes. Appl Environ Microbiol 69:5045–5050

    Article  PubMed  CAS  Google Scholar 

  • Segura A, Godoy P, van Dillewijn P, Hurtado A, Arroyo N, Santacruz S, Ramos JL (2005) Proteomic analysis reveals the participation of energy- and stress-related proteins in the response of Pseudomonas putida DOT-T1E to toluene. J Bacteriol 187:5937–5945

    Article  PubMed  CAS  Google Scholar 

  • Sondossi M, Sylvestre M, Ahmad D (1992) Effects of chlorobenzoate transformation on the Pseudomonas testosteroni biphenyl and chlorobiphenyl degradation pathway. Appl Environ Microbiol 58:485–495

    PubMed  CAS  Google Scholar 

  • Swanson CR (1969) The benzoic acid herbicides. In: Kearby PC, Kaufman DD (eds) Degradation of herbicides. Marcel Dekker, New York, pp 299–320

    Google Scholar 

  • Vrana B, Decorová K, Baláz S, Sevciková A (1996) Effect of chlorobenzoates on the degradation of polychlorinated biphenyls (PCB) by Pseudomonas stutzeri. J Microbiol Biotechnol 12:323–326

    Article  Google Scholar 

Download references

Acknowledgments

M.S. gratefully acknowledges support from the grants: FONDECYT 1070507, 1020221 and 7020221, USM 130322, 130522 and MILENIO P04/007 (MIDEPLAN). We thank Daniela Regenhardt, Rita Getzlaff and Michael Kieβ for protein analyses, Manfred Nimtz for mass spectrometry analysis, Hermann Heipieper, Edward Moore and Beatriz Cámara for helpful suggestions and critical reading of the manuscript.

Author information

Author notes

  1. Paula Martínez

    Present address: Department of Bioremediation, Helmholtz Centre for Environmental Research-UFZ, Permoserstrasse 15, 04318, Leipzig, Germany

Authors and Affiliations

  1. Laboratorio de Microbiología Molecular y Biotecnología Ambiental, Departamento de Química, Millennium Nucleus of Microbial Ecology and Environmental Microbiology and Biotechnology, Universidad Técnica Federico Santa María, Av. España 1680, Valparaíso, Chile

    Paula Martínez, Loreine Agulló, Marcela Hernández & Michael Seeger

Authors
  1. Paula Martínez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Loreine Agulló
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marcela Hernández
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Michael Seeger
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michael Seeger.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Martínez, P., Agulló, L., Hernández, M. et al. Chlorobenzoate inhibits growth and induces stress proteins in the PCB-degrading bacterium Burkholderia xenovorans LB400. Arch Microbiol 188, 289–297 (2007). https://doi.org/10.1007/s00203-007-0247-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00203-007-0247-4

Keywords

search

Navigation