Skip to main content
SpringerLink
Log in

Ion-selective electrode integrated in small-scale bioreactor for continuous intracellular pH determination in Lactobacillus plantarum

  • Original Article
  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

The aim of the present study was to develop an ion-selective electrode method for the continuous determination of the intracellular pH in Lactobacillus plantarum using a small-scale bioreactor. This method employed a salicylate-selective electrode basing on the distribution of salicylic acid across the cytoplasmic membrane. This developed electrode responded to salicylate concentrations above 20 μmol/L with a Nernstian sensitivity. The energized and concentrated cells were added into a thermostated small-scale bioreactor that contained the salicylate anions dissolved in a 100 mmol/L potassium phosphate buffer at different pH values. The changes in salicylate concentration that occurred in the medium containing bacterial suspension were measured as a voltage change. The cells of Lactobacillus plantarum showed maintenance of pH homeostasis at the studied pH ranging from 4.0 to 7.0, and they kept a neutral intracellular pH up to 5.8. The simplicity of the measuring preparation and the relatively low cellular concentration, as well as the advantages of the small-scale bioreactor, lead us to believe that the described method can facilitate the study of the physicochemical factors on the intracellular pH of lactic acid bacteria using a single pH probe in one method.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Alwazeer D, Riondet C, Cachon R (2018) Comparison between fluorescent probe and ion-selective electrode methods for intracellular pH determination in Leuconostoc mesenteroides. Curr Microbiol 75:1493–1497. https://doi.org/10.1007/s00284-018-1550-9

    Article  CAS  PubMed  Google Scholar 

  • Belguendouz T, Cachon R, Diviès C (1997) pH homeostasis and citric acid utilization: differences between Leuconostoc mesenteroides and Lactococcus lactis. Curr Microbiol 35:233–236

    Article  CAS  Google Scholar 

  • Booth IR (1985) Regulation of cytoplasmic pH in bacteria. Microbiol Rev 49:359–378

    Article  CAS  Google Scholar 

  • Bouix M, Ghorbal S (2015) Rapid assessment of Oenococcus oeni activity by measuring intracellular pH and membrane potential by flow cytometry, and its application to the more effective control of malolactic fermentation. Int J Food Microbiol 193:139–146

    Article  CAS  Google Scholar 

  • Breeuwer P, Drocourt JL, Rombouts FM, Abee T (1996) A novel method for continuous determination of the intracellular pH in bacteria with the internally conjugated fluorescent probe 5 (and 6-) -carboxyfluorescein succinimidyl ester. Appl Environ Microbiol 62:178–183

    Article  CAS  Google Scholar 

  • Cachon R, Antérieux P, Diviès C (1998) The comparative behavior of Lactococcus lactis in free and immobilized culture processes. J Biotechnol 63:211–218

    Article  CAS  Google Scholar 

  • Cook GM, Russell JB (1994) The effect of extracellular pH and lactic acid on pH homeostasis in Lactococcus lactis and Streptococcus bovis. Curr Microbiol 28:165–168

    Article  CAS  Google Scholar 

  • Glaasker E, Konings WN, Poolman B (1996) The application of pH-sensitive fluorescent dyes in lactic acid bacteria reveals distinct extrusion systems for unmodified and conjugated dyes. Mol Membr Biol 13:173–181

    Article  CAS  Google Scholar 

  • Hache C, Cachon R, Wache Y et al (1999) Influence of lactose-citrate co-metabolism on the differences of growth and energetics in Leuconostoc lactis, Leuconostoc mesenteroides ssp. mesenteroides and Leuconostoc mesenteroides ssp. cremoris. Syst Appl Microbiol 22:507–513

    Article  CAS  Google Scholar 

  • Hansen G, Johansen CL, Marten G, Wilmes J, Jespersen L, Arneborg N (2016) Influence of extracellular pH on growth, viability, cell size, acidification activity, and intracellular pH of Lactococcus lactis in batch fermentations. Appl Microbiol Biotechnol 100:5965–5976

    Article  CAS  Google Scholar 

  • Hellingwerf KJ, van Hoorn P (1985) A polyvinylchloride-membrane based anion selective electrode for continuous registration of delta pH (interior alkaline) with salicylate as the indicator probe. J Biochem Biophys Methods 11:83–93

    Article  CAS  Google Scholar 

  • Huang R, Pan M, Wan C, Shah NP, Tao X, Wei H (2016) Physiological and transcriptional responses and cross protection of Lactobacillus plantarum ZDY2013 under acid stress. J Dairy Sci 99:1002–1010. https://doi.org/10.3168/jds.2015-9993

    Article  CAS  PubMed  Google Scholar 

  • Kashket ER (1987) Bioenergetics of lactic acid bacteria: cytoplasmic pH and osmotolerance. FEMS Microbiol Rev 46:233–244

    Article  CAS  Google Scholar 

  • Kashket ER (1985) The proton motive force in bacteria: a critical assessment of methods. Annu Rev Microbiol 39:219–242

    Article  CAS  Google Scholar 

  • Kudo H, Sasaki Y (2019) Intracellular pH determination for the study of acid tolerance of lactic acid bacteria. In: Lactic Acid Bacteria. Humana Press, New York, pp 33–41

    Google Scholar 

  • Loubiere P, Salou P, Leroy MJ, Lindley ND, Pareilleux A (1992) Electrogenic malate uptake and improved growth energetics of the malolactic bacterium Leuconostoc oenos grown on glucose-malate mixtures. J Bacteriol 174:5302–5308

    Article  CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randell RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    CAS  Google Scholar 

  • Marty-Teysset C, Posthuma C, Lolkema JS, Schmitt P, Divies C, Konings WN (1996) Proton motive force generation by citrolactic fermentation in Leuconostoc mesenteroides. J Bacteriol 178:2178–2185

    Article  CAS  Google Scholar 

  • McDonald LC, Fleming HP, Hassan HM (1990) Acid tolerance of Leuconostoc mesenteroides and Lactobacillus plantarum. Appl Environ Microbiol 56:2120–2124

    Article  CAS  Google Scholar 

  • Mercade M, Lindley ND, Loubiere P (2000) Metabolism of Lactococcus lactis subsp. cremoris MG 1363 in acid stress conditions. Int J Food Microbiol 55:161–165

    Article  CAS  Google Scholar 

  • Molina-Gutierrez A, Stippl V, Delgado A, Gänzle MG, Vogel RF (2002) In situ determination of the intracellular pH of Lactococcus lactis and Lactobacillus plantarum during pressure treatment. Appl Environ Microbiol 68:4399–4406

    Article  CAS  Google Scholar 

  • Nannen NL, Hutkins RW (1991) Proton-translocating adenosine triphosphatase activity in lactic acid bacteria. J Dairy Sci 74:747–751

    Article  CAS  Google Scholar 

  • Padan E, Schuldiner S (1986) Intracellular pH regulation in bacterial cells. Methods Enzymol 125:337–352

    Article  CAS  Google Scholar 

  • Parish ME, Higgins D (1988) Isolation and identification of lactic bacteria from samples of citrus molasses and unpasteurized orange juice. J Food Sci 53:645–646

    Article  Google Scholar 

  • Paulsen IT, Brown MH, Skurray RA (1996) Proton-dependent multidrug efflux systems. Microbiol Rev 60:575–608

    Article  CAS  Google Scholar 

  • Rault A, Bouix M, Béal C (2009) Fermentation pH influences the physiological-state dynamics of Lactobacillus bulgaricus CFL1 during pH-controlled culture. Appl Environ Microbiol 75:4374–4381

    Article  CAS  Google Scholar 

  • Rottenberg H (1979) The measurement of membrane potential and deltapH in cells, organelles, and vesicles. Methods Enzymol 55:547–569

    Article  CAS  Google Scholar 

  • Rupprecht C, Wingen M, Potzkei J, Gensch T, Jaeger KE, Drepper T (2017) A novel FbFP-based biosensor toolbox for sensitive in vivo determination of intracellular pH. J Biotechnol 258:25–32

    Article  CAS  Google Scholar 

  • Schäferling M (2016) Nanoparticle-based luminescent probes for intracellular sensing and imaging of pH. Wiley Interdiscip Rev Nanomed Nanobiotechnol 8:378–413

    Article  Google Scholar 

  • Setty OH, Hendler RW, Shrager RI (1983) Simultaneous measurements of proton motive force, delta pH, membrane potential, and H+/O ratios in intact Escherichia coli. Biophys J 43:371–381

    Article  CAS  Google Scholar 

  • Tseng CP, Tsau JL, Montville TJ (1991) Bioenergetic consequences of catabolic shifts by Lactobacillus plantarum in response to shifts in environmental oxygen and pH in chemostat cultures. J Bacteriol 173:4411–4416

    Article  CAS  Google Scholar 

  • Wouters PC, Glaasker E, Smelt JPPM (1998) Effects of high pressure on inactivation kinetics and events related to proton efflux in Lactobacillus plantarum. Appl Environ Microbiol 64:509–514

    Article  CAS  Google Scholar 

  • Wu C, He G, Zhang J (2014) Physiological and proteomic analysis of Lactobacillus casei in response to acid adaptation. J Ind Microbiol Biotechnol 41:1533–1540

    Article  CAS  Google Scholar 

  • Wu C, Zhang J, Du G, Chen J (2013) Aspartate protects Lactobacillus casei against acid stress. Appl Microbiol Biotechnol 97:4083–4093

    Article  CAS  Google Scholar 

  • Xu J, Koyanagi Y, Isogai E, Nakamura S (2019) Effects of fermentation products of the commensal bacterium Clostridium ramosum on motility, intracellular pH, and flagellar synthesis of enterohemorrhagic Escherichia coli. Arch Microbiol 201:841–846

    Article  CAS  Google Scholar 

  • Zhang J, Wu C, Du G, Chen J (2012) Enhanced acid tolerance in Lactobacillus casei by adaptive evolution and compared stress response during acid stress. Biotechnol Bioprocess Eng 17:283–289. https://doi.org/10.1007/s12257-011-0346-6

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Research Center for Redox Applications in Foods (RCRAF), Department of Food Engineering, Faculty of Engineering, Iğdır University, 76000, Iğdır, Turkey

    Duried Alwazeer

  2. UMR Procédés Alimentaires et Microbiologiques, AgroSup Dijon, 1 Esplanade Erasme, 21000, Dijon, France

    Rémy Cachon

Authors
  1. Duried Alwazeer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rémy Cachon
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Duried Alwazeer.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Alwazeer, D., Cachon, R. Ion-selective electrode integrated in small-scale bioreactor for continuous intracellular pH determination in Lactobacillus plantarum. Folia Microbiol 65, 467–473 (2020). https://doi.org/10.1007/s12223-019-00749-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12223-019-00749-5

Search

Navigation