Skip to main content
Log in

Effect of water activity and protective solutes on growth and subsequent survival to air-drying of Lactobacillus and Bifidobacterium cultures

  • Applied microbial and cell physiology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Probiotic cultures of Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus casei and Lactobacillus acidophilus were grown in media having water activities (a w) adjusted between 0.99 and 0.94 with NaCl or with a mixture of glycerol and sucrose in order to find conditions of osmotic stress which would still allow for good growth. Cultures grown at a w = 0.96 or 0.99 were then recovered by centrifugation, added to a sucrose–phosphate medium and air-dried. In some assays, a 2-h osmotic stress was applied to the cell concentrate prior to air-drying. Assays were also carried out where betaine, glutamate and proline (BGP) supplements were added as protective compounds to the growth or drying media. For most strains, evidence of osmotic stress and benefits of BGP supplementation on growth occurred at a w = 0.96. Growing the cells in complex media adjusted at a w = 0.96 did not enhance their subsequent survival to air-drying, but applying the 2-h osmotic stress did. Addition of the BGP supplements to the growth medium or in the 2-h stress medium did not enhance survival to air-drying. Furthermore, addition of BGP to a sucrose–phosphate drying medium reduced survival of the cultures to air-drying. This study provides preliminary data for producers of probiotics who wish to use air-drying in replacement of freeze-drying for the stabilization of cultures.

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

Access this article

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Baliarda A, Robert H, Jebbar M, Blanco C, Deschamps A, Le Marrec C (2003) Potential osmoprotectants for the lactic acid bacteria Pediococcus pentosaceus and Tetragenococcus halophila. Int J Food Microbiol 84:13–20

    Article  CAS  Google Scholar 

  • Bayrock D, Ingledew WM (1997) Mechanism of viability loss during fluidized bed drying of baker’s yeast. Food Res Int 30:417–425

    Article  Google Scholar 

  • Carvalho AS, Silva J, Ho P, Teixeira P, Malcata FX, Gibbs P (2003) Protective effect of sorbitol and monosodium glutamate during storage of freeze-dried lactic acid bacteria. Lait 83:203–210

    Article  CAS  Google Scholar 

  • Champagne CP, Gardner N (2001) The effect of protective ingredients on the survival of immobilized cells of Streptococcus thermophilus to air- and freeze-drying. Electron J Biotechnol Vol 4, No 3, http://ejb.ucv.cl/content/ vol4/ issue3/ full/ 4/ index.html. Issue of Dec 15, 2001

  • Champagne CP, Ross P, Saarela M, Flemming Hansen K, Charalampopoulos D (2011) Recommendations for the viability assessment of probiotics in cultured concentrates and food matrices. Int J Food Microbiol 149:185–193

    Article  Google Scholar 

  • De Angelis M, Gobbetti M (2004) Environmental stress responses in Lactobacillus. A review. Proteomics 4:106–122

    Article  Google Scholar 

  • De Man JC, Rogosa M, Sharpe ME (1960) A medium for the cultivation of lactobacilli. J Appl Bacteriol 23:130–135

    Article  Google Scholar 

  • Glaasker E, Konings WN, Poolman B (1996) Glycine betaine fluxes in Lactobacillus plantarum during osmostasis and hyper-and hypo-osmotic shock. J Biol Chem 271:10060–11006

    Article  CAS  Google Scholar 

  • Glaasker E, Tjan FSB, Steeg PFT, Konings WN, Poolman B (1998) Physiological response of Lactobacillus plantarum to salt and nonelectrolyte stress. J Bacteriol 180:4718–4723

    CAS  Google Scholar 

  • Gouesbet G, Jan G, Boyaval P (2001) Hosts, environment, stress, phages and Lactobacillus delbrueckii ssp. bulgaricus thermotolerance. Lait 81:301–309

    Article  CAS  Google Scholar 

  • Guillot A, Obis D, Mistou MY (2000) Fatty acid membrane composition and activation of glycine-betaine transport in Lactococcus lactis subjected to osmotic stress. Int J Food Microbiol 55:47–51

    Article  CAS  Google Scholar 

  • Hutkins RW, Ellefson WL, Hashket ER (1987) Betaine transport imparts osmotolerance on a strain of Lactobacillus acidophilus. Appl Environ Microbiol 53:2275–2281

    CAS  Google Scholar 

  • Ishibashi N, Tatematsu T, Shimamura S, Tomota M, Okonogi S (1985) Effect of water activity on the viability of freeze-dried bifidobacteria and lactic acid bacteria. Fundamentals and application of freeze drying to biological materials, dyes and foodstuffs. Int Inst Refrig, Paris, pp 227-232.

  • Kets EPW, De Bont JAM (1994) Protective effect of betaine on survival of Lactobacillus plantarum subjected to drying. FEMS Microbiol Lett 116:251–256

    Article  CAS  Google Scholar 

  • Kets EPW, Teunissen PJM, De Bont JAM (1996) Effect of compatible solutes on survival of lactic acid bacteria subjected to drying. Appl Environ Microbiol 62:259–261

    CAS  Google Scholar 

  • King VAE, Su JT (1993) Dehydration of Lactobacillus acidophilus. Process Biochem 28:47–52

    Article  CAS  Google Scholar 

  • Larsen RF, Añón MC (1990) Effect of water activity of milk upon growth and acid production by mixed cultures of Streptococcus thermophilus and Lactobacillus bulgaricus. J Food Sci 55(708–710):800

    Google Scholar 

  • Le Marrec C, Bon E, Lonvaud-Funel A (2007) Tolerance to high osmolality of the lactic acid bacterium Oenococcus oeni and identification of potential osmoprotectants. Int J Food Microbiol 115:335–342

    Article  Google Scholar 

  • Lievense LC, Verbeek MAM, Meerdink G, van’t Riet K (1990) Inactivation of Lactobacillus plantarum during drying. II. Measurement and modelling of the thermal inactivation. Biosepar 1:161–170

    CAS  Google Scholar 

  • Linders LJM, de Jong GIW, Meerdink G, van’t Riet K (1997a) Carbohydrates and the dehydration inactivation of Lactobacillus plantarum: the role of moisture distribution and water activity. J Food Engin 31:237–250

    Article  Google Scholar 

  • Linders LJM, Meerdink G, van’t Riet K (1997b) Effect of growth parameters on the residual activity of Lactobacillus plantarum after drying. J Appl Microbiol 82:683–688

    Article  Google Scholar 

  • Marcotte M, Brodeur C (2001) Le séchage? Pas si aride. Le Monde Aliment 5:20–22

    Google Scholar 

  • Meiners JA, Boquel D (2009) Applications of multi wall probiotic encapsulation in dairy products. Agro Food Ind Hi-tech 20:14–16

    CAS  Google Scholar 

  • Mille Y, Obert JP, Beney L, Gervais P (2004) New drying process for lactic bacteria based on their dehydration behaviour in liquid medium. Biotechnol Bioengin 88:71–76

    Article  CAS  Google Scholar 

  • Molenaar D, Hagting A, Alkema H, Driessen AJM, Konings WN (1993) Characteristics and osmoregulatory roles of uptake systems for proline and glycine betaine in Lactococcus lactis. J Bacteriol 175:5438–5444

    CAS  Google Scholar 

  • O’Callaghan J, Condon S (2000) Growth of Lactococcus lactis strains at low water activity: correlation with the ability to accumulate glycine betaine. Int J Food Microbiol 55:127–131

    Article  Google Scholar 

  • Paul E, Fages J, Blanc P, Goma G, Pareilleux A (1993) Survival of alginate-entrapped cells of Azospirillum lipoferum during dehydration and storage in relation to water properties. Appl Microbiol Biotechnol 40:34–39

    Article  CAS  Google Scholar 

  • Piuri M, Sanchez-Rivas C, Ruzal SM (2003) Adaptation to high salt in Lactobacillus: role of peptides and proteolytic enzymes. J Appl Microbiol 95:372–379

    Article  CAS  Google Scholar 

  • Poirier I, Maréchal PA, Evrard C, Gervais P (1998) Escherichia coli and Lactobacillus plantarum responses to osmotic stress. Appl Microbiol Biotechnol 50:704–709

    Article  CAS  Google Scholar 

  • Prasad J, McJarrow P, Gopal P (2003) Heat and osmotic stress response of probiotic Lactobacillus rhamnosus HN001 (DR20) in relation to viability after drying. Appl Environ Microbiol 69:917–925

    Article  CAS  Google Scholar 

  • Romeo Y, Bouvier J, Gutierrez C (2001) La réponse au stress osmotique des bactéries lactiques Lactococcus lactis et Lactobacillus plantarum (mini-revue). Lait 81:49–55

    Article  CAS  Google Scholar 

  • Saarela M, Virkajarvi I, Alakomi HL, Mattila-Sandholm T, Vaari A, Suomalainen T, Matto J (2005) Influence of fermentation time, cryoprotectant and neutralization of cell concentrate on freeze-drying survival, storage stability, and acid and bile exposure of Bifidobacterium animalis ssp. lactis cells produced without milk-based ingredients. J Appl Microbiol 99:1330–1339

    Article  CAS  Google Scholar 

  • Sanders JW, Venema G, Kok J (1999) Environmental stress responses in Lactococcus lactis. FEMS Microbiol Rev 23:483–501

    Article  CAS  Google Scholar 

  • Santivarangkna C, Wenning M, Foerst P, Kulozik U (2007) Damage of cell envelope of Lactobacillus helveticus during vacuum drying. J Appl Microbiol 102:748–756

    Article  CAS  Google Scholar 

  • Selmer-Olsen E, Sorhaug T, Birkeland SE, Pehrson R (1999) Survival of Lactobacillus helveticus entrapped in Ca-alginate in relation to water content, storage and rehydration. J Ind Microbiol Biotechnol 23:79–85

    Article  CAS  Google Scholar 

  • Silva J, Carvalho AS, Ferreira R, Vitorino R, Amado F, Domingues P, Teixeira P, Gibbs PA (2005) Effect of the pH of growth on the survival of Lactobacillus delbrueckii subsp. bulgaricus to stress conditions during spray-drying. J Appl Microbiol 98:775–782

    Article  CAS  Google Scholar 

  • Troller JA, Stinson JV (1981) Moisture requirements for growth and metabolite production by lactic acid bacteria. Appl Environ Microbiol 42:682–687

    CAS  Google Scholar 

  • Turker EN, Hamamci H (1998) Storage behaviour of immobilized dried micro-organisms. Food Microbiol 15:3–11

    Article  Google Scholar 

Download references

Acknowledgments

Gratitude is expressed to Anais Besnier and Leila El Alaoui for their technical contributions.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Claude P. Champagne.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Champagne, C.P., Raymond, Y. & Simon, JP. Effect of water activity and protective solutes on growth and subsequent survival to air-drying of Lactobacillus and Bifidobacterium cultures. Appl Microbiol Biotechnol 95, 745–756 (2012). https://doi.org/10.1007/s00253-012-3912-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-012-3912-8

Keywords

Navigation