Skip to main content
SpringerLink
Log in

Viability and Stress Response of Putative Probiotic Lactobacillus plantarum Strains in Honey Environment

  • Published:
Probiotics and Antimicrobial Proteins Aims and scope Submit manuscript

Abstract

Due to problem of preservation of dairy products which serve as a matrix for probiotics, it is challenging to use these probiotics as food supplements in many developing countries. To determine the suitability of the Lactobacillus strains for exploitation as probiotics in honey, we investigated the effect of their storage on the viability, functionality, and the mechanism associated with their protective effect. Three isolates obtained from our laboratory collection were identified through amplification of the 16S rRNA gene. The viability of the strains in honey at different storage conditions was studied. Three genes (hdc, gtf, and clpL) responsible for the resistance of bacteria in acidic environments were screened. SDS-PAGE analysis of total protein was performed to observe protein profile changes of the strains after exposure to honey. All the three isolates, namely, GGU, GLA51, and GLP56, were identified as Lactobacillus plantarum strains. After 28 days of storage in honey at 4 °C, viable cell concentrations of the three strains were higher than 2.04 × 106 CFU/ml. During the same period at room temperature, only the Lactobacillus plantarum GLP56 strain remained viable with a cell concentration of 1.86 × 104 CFU/ml. The clpL gene coding for ATPase was detected in all the three strains. The protein of molecular weight ~ 50 kDa was absent in the protein profile of Lactobacillus plantarum GGU after 60 days of storage in honey at 4 °C. The Lactobacillus plantarum GLP56, Lactobacillus plantarum GLA51, and Lactobacillus plantarum GGU strains exposed to honey can withstand acidic environmental stress but their viability declines over time.

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
Fig. 3

Similar content being viewed by others

References

  1. Tripathi MK, Giri SK (2014) Probiotic functional foods: survival of probiotics during processing and storage. J Funct Foods 9:225–241. https://doi.org/10.1016/j.jff.2014.04.030

    Article  CAS  Google Scholar 

  2. Shukla G, Sharma G, Goyal N (2010) Probiotic characterization of lactobacilli and yeast strains isolated from whey beverage and therapeutic potential of lactobacillus yoghurt in murine giardiasis. Am J Biomed Sci 2:248–261

    Article  Google Scholar 

  3. Bahr F, Lejeune A, Dubois-Dauphin R, Elmejdoub T, Boulahrouf A, Thonart P (2014) Characterization of Lactobacillus strains isolated from Algerian children faeces for their probiotic properties. Afr J Microbiol Res 8:297–303

    Article  Google Scholar 

  4. Folarin A, Oguntoyinbo NA (2015) Multifunctional properties of Lactobacillus plantarum strains isolated from fermented cereal foods. J Funct Foods 17:621–631

    Article  Google Scholar 

  5. Yazdi MKS, Davoodabadi A, Zarin HRK, Ebrahimi MT, Dallal MMS (2017) Characterisation and probiotic potential of lactic acid bacteria isolated from Iranian traditional yogurts. Ital J Anim Sci 16(2):185–188. https://doi.org/10.1080/1828051X.2016.1222888

    Article  CAS  Google Scholar 

  6. Perricome M, Bevilacqua A, Altieri C, Sinigaglia M, Corbo MR (2015) Challenges for the production of probiotic fruit juices. Beverages 1(2):95–103. https://doi.org/10.3390/beverages1020095

    Article  CAS  Google Scholar 

  7. Perricome M, Corbo MR, Sinigaglia M, Speranza B, Bevilacqua A (2014) Viability of Lactobacillus reuteriin fruit juices. J Funct Foods 10:421–426. https://doi.org/10.1016/j.jff.2014.07.020

    Article  CAS  Google Scholar 

  8. Hassan AA, Aly MM, El-Hadidie ST (2012) Production of cereal-based probiotic beverages. World Appl Sci J 19:1367–1380

    CAS  Google Scholar 

  9. Vasudha S, Mishra HN (2013) Non dairy probiotic beverages. Int Food Res J 20:7–15

    CAS  Google Scholar 

  10. Nath AH, Ukkuru MP, Kumari MK (2015) Development of a probiotic honey beverage. Int J Appl Pure Sci Agric 9:1–9

    CAS  Google Scholar 

  11. Heenan CN, Adams C, Hoskena RW, Fleet H (2004) Survival and sensory acceptability of probiotic microorganisms in a non fermented frozen vegetarian dessert. LWT-Food Sci Technol 37(4):461–466. https://doi.org/10.1016/j.lwt.2003.11.001

    Article  CAS  Google Scholar 

  12. Granato D, Branco GF, Nazzaro F, Cruz AG, Faria JA (2010) Functional foods and non dairy probiotic food development: trends, concepts, and products. Compr Rev Food Sci Food Saf 9(3):292–302. https://doi.org/10.1111/j.1541-4337.2010.00110.x

    Article  CAS  Google Scholar 

  13. Bogdanov S, Gallmann P, Stangaciu SC, Herbuliez T (2006) Produits apicoles et santé. Station de recherche, Agroscope Liebefeld-Posieux, ALP, N° 41f. ISSN 1661–0660, p 4–18

  14. Gibson GR, Roberfroid MB (1995) Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr 12:1401–1412

    Google Scholar 

  15. Pandiyan C, Annal VR, Kumaresan G, Murugan B, Gopalakrishnamurthy TR (2012) In vivo and in vitro effect of Lactobacillus acidophilus in synbiotic ice cream enriched with whey protein concentrate. Int Food Res J 19:441–446

    CAS  Google Scholar 

  16. Shin HS, Ustunol Z (2005) Carbohydrate composition of honey from different floral sources and their influence on growth of selected intestinal bacteria: an in vitro comparison. Food Res Int 38(6):721–728. https://doi.org/10.1016/j.foodres.2005.01.007

    Article  CAS  Google Scholar 

  17. Shamala TR, Shri JY, Saibaba P (2000) Stimulatory effect of honey on multiplication of lactic acid bacteria under in vitro and in vivo conditions. Lett Appl Microbiol 30(6):453–455. https://doi.org/10.1046/j.1472-765x.2000.00746.x

    Article  CAS  PubMed  Google Scholar 

  18. Bemmo KUL, Sahoo M, Jayabalan R, Zambou NF (2016) Honey, probiotics and prebiotics: review. Res J Pharm Biol Chem Sci 7:2428–2438

    Google Scholar 

  19. Kwakman PH, Te Velde AA, De Boer L (2010) How honey kills bacteria. Fed Am Soc Exp Biol J 7:2576–2581

    Google Scholar 

  20. Bogdanov S, Haldimann M, Luginbühl W, Gallmann P (2007) Minerals in honey: environmental, geographical and botanical aspects. J Apic Res Bee World 4:269–275

    Article  Google Scholar 

  21. Wu R, Xu X, Meng L, Zou T, Tang X, Wu J, Yue X (2014) Identification of salt stress responsive protein in Lactobacillus Paracasei LN-1 using SDS-PAGE. IERI Procedia 8:60–65. https://doi.org/10.1016/j.ieri.2014.09.011

    Article  Google Scholar 

  22. Sugimoto S, Abdullah-Al-Mahin SK (2008) Molecular chaperones in lactic acid bacteria: physiological consequences and biochemical properties. J Biosci Bioeng 106(4):324–336. https://doi.org/10.1263/jbb.106.324

    Article  CAS  PubMed  Google Scholar 

  23. Mills S, Stanton C, Fitzgerald GF, Ross RP (2011) Enhancing the stress responses of probiotics for a lifestyle from gut to product and back again. Microb Cell Factories 10:1–15

    Article  Google Scholar 

  24. Corcoran BM, Stanton C, Fitzgerald G, Ross RP (2008) Life under stress: the probiotic stress response and how it may be manipulated. Curr Pharm Des 14(14):1382–1399. https://doi.org/10.2174/138161208784480225

    Article  CAS  PubMed  Google Scholar 

  25. Sieladie DV (2012) Potentiel probiotique des lactobacilles isolés du lait de vache dans les hautes terres de l’ouest du Cameroun. Thèse de Doctorat/PhD. Université de Dschang, Faculté des sciences, Département de Biochimie, p 62–100

  26. Costantini A, Cersosimo M, Del Prete V, Garcia-Moruno E (2006) Production of biogenic amines by lactic acid bacteria: screening by PCR, thin-layer chromatography, and high-performance liquid chromatography of strains isolated from wine and must. J Food Prot 69(2):391–396. https://doi.org/10.4315/0362-028X-69.2.391

    Article  CAS  PubMed  Google Scholar 

  27. Stack HM, Kearney N, Stanton C, Fitzgerald GF, Ross RP (2010) Association of beta-glucan endogenous production with increased stress tolerance of intestinal lactobacilli. Appl Environ Microbiol 76(2):500–507. https://doi.org/10.1128/AEM.01524-09

    Article  CAS  PubMed  Google Scholar 

  28. Vrancken G, Rimaux T, Weckx S, De Vuyst L, Leroy F (2009) Environmental pH determines citrulline and ornithine release through the arginine deiminase pathway in lactobacillus fermentum. Int J FoodMicrobiol 135(3):216–222. https://doi.org/10.1016/j.ijfoodmicro.2009.07.035

    Article  CAS  Google Scholar 

  29. IUPAC (1979) Standard methods for the Analysis of oils, fats and derivatives 151p. ISBN-0-632-03337-1

  30. Association of Official Analytical Chemists (1990) In: Helrich K (ed) Official methods of analysis, 15th ed. Arlington, VA USA, p 10251026–10331034

  31. Bogdanov S, Martin P, Lüllmann C, Borneck R et al (1997) Harmonised methods of the European honey commission. Apidologie 29:513–524

    Article  Google Scholar 

  32. Zambou NF, Moumbe FG, Kaktcham PM (2013) Antimicrobial activity of probiotic strain Lactobacillus Plantarum isolated from “Sha’a” and assessment of its viability in local honey. J Microbiol Biotechnol Food Sci 3:226–231

    Google Scholar 

  33. Riazi A, Ziar H (2012) Effect of honey and starter culture on growth, acidification, sensory properties and bifidobacteria cell counts in fermented skimmed milk. Afr J Microbiol Res 6:486–498

    CAS  Google Scholar 

  34. Jan MS, Mohd NM, Norrakiah AS (2010) Fructooligosaccharides in honey and effects of honey on growth of Bifidobacterium longum BB 536. Int Food Res J 17:557–561

    Google Scholar 

  35. Chick H, Shin HS, Ustunol Z (2001) Growth and acid production by lactic acid bacteria and bifidobacteria grown in skim milk containing honey. J Food Sci 66(3):478–481. https://doi.org/10.1111/j.1365-2621.2001.tb16134.x

    Article  CAS  Google Scholar 

  36. De Man JC, Rogosa M, Sharpe ME (1960) A medium for the cultivation of Lactobacilli. J Appl Bact 23(1):130–135. https://doi.org/10.1111/j.1365-2672.1960.tb00188.x

    Article  Google Scholar 

  37. Prasad J, Gill H, Smart J, Gopal PK (1998) Selection and characterization of Lactobacillus and Bifidobacterium strains for use as probiotic. Int Dairy J 8(12):993–1002. https://doi.org/10.1016/S0958-6946(99)00024-2

    Article  Google Scholar 

  38. Oluwajoba SO, Akinyosoye FA, Oyetayo VO (2013) In vitro screening and selection of probiotic lactic acid bacteria isolated from spontaneously fermenting Kunu-Zaki. Adv Microbiol 3(04):309–316. https://doi.org/10.4236/aim.2013.34044

    Article  CAS  Google Scholar 

  39. Cotter PD, Hill C (2003) Surviving the acid test: responses of gram positive bacteria to low pH. Microbiol Mol Biol Rev 67(3):429–453. https://doi.org/10.1128/MMBR.67.3.429-453.2003

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Coton E, Rollan GC, Lonvaud-Funel A (1998) HistidinecarboxylaseofLeuconostocoenos9204: purification, kinetic properties, cloning and nucleotide sequence of the hdc gene. J Appl Microbiol 84(2):143–151. https://doi.org/10.1046/j.1365-2672.1998.00271.x

    Article  CAS  PubMed  Google Scholar 

  41. Pessione E, Cirrincione S (2016) Bioactive molecules released in food by lactic acid bacteria: encrypted peptides and biogenic amines. Front Microbiol 7:1–19

    Article  Google Scholar 

  42. Turpin W, Humblot C, Guyot JP (2011) Genetic screening of functional properties of lactic acid bacteria in a fermented pearl millet slurry and in the metagenome of fermented starchy foods. Appl Environ Microbiol 77(24):8722–8734. https://doi.org/10.1128/AEM.05988-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Wall T, Båth K, Britton RA, Jonsson H, Versalovic J, Roos S (2007) The early response to acid shock in Lactobacillus reuteri involves the ClpL chaperone and a putative cell wall-altering esterase. Appl Environ Microbiol 73(12):3924–3935. https://doi.org/10.1128/AEM.01502-06

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Rossano R, Larocca M, Polito T, Perna AM, Padula MC, Martelli G, Riccio P (2012) What are the proteolytic enzymes of honey and what they do tell us? A fingerprint analysis by 2-D Zymography of unifloral honeys. PLoS One 7(11):e49164. https://doi.org/10.1371/journal.pone.0049164

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Brudzynski K, Sjaarda C (2015) Honey glycoproteins containing antimicrobial peptides, jelleins of the major royal jelly protein 1, are responsible for the cell wall lytic and bactericidal activities of honey. PLoS One 10(4):e0120238. https://doi.org/10.1371/journal.pone.0120238

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Hamon E, Horvatovich P, Izquierdo E, Bringel F, Aoudé-Werner D, Ennahar S (2011) Comparative proteomic analysis of Lactobacillus plantarum for the identification of key proteins in bile tolerance. BMC Microbiol 11:191–201

    Article  Google Scholar 

Download references

Acknowledgements

The authors are grateful for the assistance and technical support of the CAS-TWAS Centre of Excellence for Biotechnology (CoEBI0) researchers.

Funding

This research was supported by Chinese Academy of Sciences Visiting President’s International Fellowship Initiative. Grant N° 2015VBB051.

Author information

Authors and Affiliations

  1. Laboratory of Biochemistry, Food Science and Nutrition (LABPMAN)—Department of Biochemistry—Faculty of Science, University of Dschang, Cameroon, P.O. Box: 67, Dschang, Cameroon

    Bemmo Kamdem Ulrich Landry & Zambou Ngoufack François

  2. CAS Key Laboratory of Microbial, Physiological and Metabolic Engineering, Institute of Microbiology, Beijing, China

    Zambou Ngoufack François, Zhu Taicheng & Yin Li

  3. Centre of Excellence for Biotechnology, CoEBIO, Institute of Microbiology, Beijing, China

    Rui-Yan Wang

Authors
  1. Bemmo Kamdem Ulrich Landry
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zambou Ngoufack François
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rui-Yan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhu Taicheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zambou Ngoufack François.

Ethics declarations

This article does not contain any studies with human or animal subjects.

Conflict of Interest

Author ZAMBOU NGOUFACK Francois has received research Grant N° 2015VBB051 from Chinese Academy of Sciences Visiting President’s International Fellowship Initiative.

Author BEMMO KAMDEM Ulrich Landry is a PhD student supervised by ZAMBOU NGOUFACK Francois. He performed some experiments during this research in the university of Dschang.

Authors Rui Yan Wang, Zhu Taicheng and Yin Li coordinated the grant and supervised research activities that were carried out at the Institute of Microbiology under the Grant N° 2015VBB051.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Landry, B.K.U., François, Z.N., Wang, RY. et al. Viability and Stress Response of Putative Probiotic Lactobacillus plantarum Strains in Honey Environment. Probiotics & Antimicro. Prot. 10, 629–637 (2018). https://doi.org/10.1007/s12602-017-9358-7

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12602-017-9358-7

Keywords

Search

Navigation