Skip to main content
Log in

Investigation of the instability and low water kefir grain growth during an industrial water kefir fermentation process

  • Biotechnological products and process engineering
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

A poorly performing industrial water kefir production process consisting of a first fermentation process, a rest period at low temperature, and a second fermentation process was characterized to elucidate the causes of its low water kefir grain growth and instability. The frozen-stored water kefir grain inoculum was thawed and reactivated during three consecutive prefermentations before the water kefir production process was started. Freezing and thawing damaged the water kefir grains irreversibly, as their structure did not restore during the prefermentations nor the production process. The viable counts of the lactic acid bacteria and yeasts on the water kefir grains and in the liquors were as expected, whereas those of the acetic acid bacteria were high, due to the aerobic fermentation conditions. Nevertheless, the fermentations progressed slowly, which was caused by excessive substrate concentrations resulting in a high osmotic stress. Lactobacillus nagelii, Lactobacillus paracasei, Lactobacillus hilgardii, Leuconostoc mesenteroides, Bifidobacterium aquikefiri, Gluconobacter roseus/oxydans, Gluconobacter cerinus, Saccharomyces cerevisiae, and Zygotorulaspora florentina were the most prevalent microorganisms. Lb. hilgardii, the microorganism thought to be responsible for water kefir grain growth, was not found culture-dependently, which could explain the low water kefir grain growth of this industrial process.

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

  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    Article  CAS  PubMed  Google Scholar 

  • Cocolin L, Bisson LF, Mills DA (2000) Direct profiling of the yeast dynamics in wine fermentations. FEMS Microbiol Lett 189:81–87

    Article  CAS  PubMed  Google Scholar 

  • D’Amore T, Panchal CJ, Russeil I, Stewart GG (1988) Osmotic pressure effects and intracellular accumulation of ethanol in yeast during fermentation. J Ind Microbiol Biotechnol 2:365–372

    Google Scholar 

  • Edwards U, Rogall T, Blocker H, Emde M, Bottger EC (1989) Isolation and direct complete nucleotide determination of entire genes—characterization of a gene coding for 16S-ribosomal RNA. Nucleic Acids Res 17:7843–7853

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ercolini D, Moschetti G, Blaiotta G, Coppola S (2001) Behavior of variable V3 region from 16S rDNA of lactic acid bacteria in denaturing gradient gel electrophoresis. Curr Microbiol 42:199–202

    Article  CAS  PubMed  Google Scholar 

  • Franzetti L, Galli A, Pagani MA, De Noni I (1998) Microbiological and chemical investigations on “sugar kefir” drink. Ann Microbiol Enzimol 48:67–80

    CAS  Google Scholar 

  • Garcia-Armisen T, Papalexandratou Z, Hendryckx H, Camu N, Vrancken G, De Vuyst L, Cornelis P (2010) Diversity of the total bacterial community associated with Ghanaian and Brazilian cocoa bean fermentation samples as revealed by a 16S rRNA gene clone library. Appl Microbiol Biotechnol 87:2281–2292

    Article  CAS  PubMed  Google Scholar 

  • Gulitz A, Stadie J, Ehrmann MA, Ludwig W, Vogel RF (2013) Comparative phylobiomic analysis of the bacterial community of water kefir by 16S rRNA gene amplicon sequencing and ARDRA analysis. J Appl Microbiol 114:1082–1091

    Article  CAS  PubMed  Google Scholar 

  • Gulitz A, Stadie J, Wenning M, Ehrmann MA, Vogel RF (2011) The microbial diversity of water kefir. Int J Food Microbiol 151:284–288

    Article  CAS  PubMed  Google Scholar 

  • Gulitz AJ (2013) Analysis of the diversity of water kefir microbiota by culture-dependent and -independent approaches. Dissertation, Technischen Universität Munchen

  • Hehre EJ (1946) Studies on the enzymatic synthesis of dextran from sucrose. J Biol Chem 163:221–233

    CAS  PubMed  Google Scholar 

  • Janssens M, Myter N, De Vuyst L, Leroy F (2012) Species diversity and metabolic impact of the microbiota are low in spontaneously acidified Belgian sausages with an added starter culture of Staphylococcus carnosus. Food Microbiol 29:167–177

    Article  CAS  PubMed  Google Scholar 

  • Laureys D, Cnockaert M, De Vuyst L, Vandamme P (2016) Bifidobacterium aquikefiri sp. nov. isolated from water kefir. Int J Syst Evol Microbiol 66:1281–1286

    Article  CAS  Google Scholar 

  • Laureys D, De Vuyst L (2014) Microbial species diversity, community dynamics, and metabolite kinetics of water kefir fermentation. Appl Environ Microbiol 80:2564–2572

    Article  PubMed  PubMed Central  Google Scholar 

  • Marsh AJ, Hill C, Ross RP, Cotter PD (2014) Fermented beverages with health-promoting potential: past and future perspectives. Trends Food Sci Technol 38:113–124

    Article  CAS  Google Scholar 

  • Marsh AJ, O’Sullivan O, Hill C, Ross RP, Cotter PD (2013) Sequence-based analysis of the microbial composition of water kefir from multiple sources. FEMS Microbiol Lett 348:79–85

    Article  CAS  PubMed  Google Scholar 

  • Moinas M, Horisberger M, Bauer H (1980) The structural organization of the Tibi grain as revealed by light, scanning and transmission microscopy. Arch Microbiol 128:157–161

    Article  Google Scholar 

  • Papalexandratou Z, Falony G, Romanens E, Jimenez JC, Amores F, Daniel HM, De Vuyst L (2011) Species diversity, community dynamics, and metabolite kinetics of the microbiota associated with traditional Ecuadorian spontaneous cocoa bean fermentations. Appl Environ Microbiol 77:7698–7714

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Papalexandratou Z, Lefeber T, Bahrim B, Lee OS, Daniel HM, De Vuyst L (2013) Hanseniaspora opuntiae, Saccharomyces cerevisiae, Lactobacillus fermentum, and Acetobacter pasteurianus predominate during well-performed Malaysian cocoa bean box fermentations, underlining the importance of these microbial species for a successful cocoa bean fermentation process. Food Microbiol 35:73–85

    Article  CAS  PubMed  Google Scholar 

  • Pidoux M (1989) The microbial flora of sugary kefir grain (the gingerbeer plant): biosynthesis of the grain from Lactobacillus hilgardii producing a polysaccharide gel. J Appl Microbiol 5:223–238

    Google Scholar 

  • Pothakos V, Illeghems K, Laureys D, Spitaels F, Vandamme P, De Vuyst L (2016) Acetic acid bacteria in fermented food and beverage ecosystems. In: Matsushita K, Toyama H, Tonouchi N, Okamoto-Kainuma A (eds) Acetic acid bacteria: ecology and physiology. Springer, Tokyo, pp 73–100

    Google Scholar 

  • Satokari RM, Vaughan EE, Akkermans ADL, Saarela M, de Vos WM (2001) Bifidobacterial diversity in human feces detected by genus-specific PCR and denaturing gradient gel electrophoresis. Appl Environ Microbiol 67:504–513

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. J Bacteriol 172:4238–4246

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • White T, Buns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis N, Gelfand D, Snisky J, White T (eds) PCR protocols: a guide to methods and applications. Academic Press, NY, pp 315–322

    Google Scholar 

Download references

Acknowledgements

The authors acknowledge their financial support of the Research Council of the Vrije Universiteit Brussel (SRP, IRP, and IOF projects), the Hercules Foundation, the Research Foundation-Flanders (FWO-Vlaanderen), and the KMO-portefeuille (Agentschap Ondernemen). DL was the recipient of a PhD fellowship of the Vrije Universiteit Brussel.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Luc De Vuyst.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

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

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Laureys, D., Van Jean, A., Dumont, J. et al. Investigation of the instability and low water kefir grain growth during an industrial water kefir fermentation process. Appl Microbiol Biotechnol 101, 2811–2819 (2017). https://doi.org/10.1007/s00253-016-8084-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-016-8084-5

Keywords

Navigation