Application of Probiotics in Folate Bio-Fortification of Yoghurt

  • Mohammad Khalili
  • Aziz Homayouni Rad
  • Ahmad Yari Khosroushahi
  • Hadi Khosravi
  • Somayeh JafarzadehEmail author


Folate deficiency is a public health concern affecting all age groups worldwide. The available evidence reveals that adding probiotic bacteria to the yoghurt starter cultures during yoghurt production process under fermentation conditions increases the folate content of yoghurt. The present study was conducted to measure two folate derivatives, i.e., 5-methyltetrahydrofolate and 5-formyltetrahydrofolate, in bio-fortified yoghurt samples including (1) yoghurt containing Streptococcus thermophilus and Lactobacillus bulgaricus, (2) probiotic yoghurt containing Lactobacillus acidophilus LA-5 and Bifidobacterium lactis BB-12, (3) probiotic yoghurt containing native strains of Lactobacillus plantarum 15HN, (4) probiotic yoghurt containing native strains of Lactococcus lactis 44Lac, and (5) probiotic yoghurt containing commercial strains of Lactobacillus plantarum LAT BY PL. During storage at 4 °C for 21 days, the highest levels of 5-methyltetrahydrofolate and 5-formyltetrahydrofolate, which were statistically significant, were detected in the yoghurt made using Lact. plantarum 15HN. Moreover, the highest total folate concentration (1487 ± 96.42 μg/L) was specified in the yoghurt containing Lact. plantarum 15HN on the 7th day. It can be conjectured that this product can be suggested as a proper alternative to synthetic folic acid and may not have the side effects of using synthetic folic acid overdoses.


Probiotic Folate Bio-fortification Yoghurt 



The study was financially supported by Research Vice Chancellor of Tabriz University of Medical Sciences.

Compliance with Ethical Standards

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.

Informed Consent

Formal consent was not required considering the type of the present study.


  1. 1.
    Rad AH, Khosroushahi AY, Khalili M, Jafarzadeh S (2016) Folate bio-fortification of yoghurt and fermented milk: a review. Dairy Sci Technol 96(4):427–441CrossRefGoogle Scholar
  2. 2.
    Laiño JE, Juarez del Valle M, Savoy de Giori G, LeBlanc JGJ (2013) Development of a high folate concentration yogurt naturally bio-enriched using selected lactic acid bacteria. LWT Food Sci Technol 54(1):1–5. CrossRefGoogle Scholar
  3. 3.
    Holasova M, Fiedlerova V, Roubal P, Pechacova M (2005) Possibility of increasing natural folate content in fermented milk products by fermentation and fruit component addition. Czech J Food Sci 23(5):196–201CrossRefGoogle Scholar
  4. 4.
    LeBlanc JG, de Giori GS, Smid EJ, Hugenholtz J, Sesma F (2007) Folate production by lactic acid bacteria and other food-grade microorganisms. In: Méndez-Vilas A (ed) Communicating current research and educational topics and trends in applied microbiology, 1st edn. FORMATEX, Oklahoma, pp 329–339Google Scholar
  5. 5.
    Papastoyiannidis G, Polychroniadou A, Michaelidou A-M, Alichanidis E (2006) Fermented milks fortified with B-group vitamins: vitamin stability and effect on resulting products. Food Sci Technol Int 12(6):521–529CrossRefGoogle Scholar
  6. 6.
    Storozhenko S, Ravanel S, Zhang G-F, Rébeillé F, Lambert W, Van Der Straeten D (2005) Folate enhancement in staple crops by metabolic engineering. Trends Food Sci Technol 16(6):271–281CrossRefGoogle Scholar
  7. 7.
    Geisel J (2003) Folic acid and neural tube defects in pregnancy: a review. J Perinat Neonatal Nur 17(4):268–279CrossRefGoogle Scholar
  8. 8.
    Rossi M, Amaretti A, Raimondi S (2011) Folate production by probiotic bacteria. Nutrients 3(1):118–134CrossRefGoogle Scholar
  9. 9.
    Taheraghdam AA, Dalirakbari N, Khalili M, Soltani M, Ahari SS (2016) Hyperhomocysteinemia, low vitamin B12, and low folic acid: are risk factors of cerebral vascular thrombosis in northwest Iran? J Res Med Sci 21:16–20CrossRefGoogle Scholar
  10. 10.
    Laiño JE, LeBlanc JG, Savoy de Giori G (2012) Production of natural folates by lactic acid bacteria starter cultures isolated from artisanal Argentinean yogurts. Can J Microbiol 58(5):581–588CrossRefGoogle Scholar
  11. 11.
    Bailey LB, Rampersaud GC, Kauwell GP (2003) Folic acid supplements and fortification affect the risk for neural tube defects, vascular disease and cancer: evolving science. J Nutr 133(6):1961S–1968SCrossRefGoogle Scholar
  12. 12.
    Bailey SW, Ayling JE (2009) The extremely slow and variable activity of dihydrofolate reductase in human liver and its implications for high folic acid intake. Proc Natl Acad Sci 106(36):15424–15429CrossRefGoogle Scholar
  13. 13.
    Baggott JE, Oster RA, Tamura T (2012) Meta-analysis of cancer risk in folic acid supplementation trials. Cancer Epidemiol 36(1):78–81CrossRefGoogle Scholar
  14. 14.
    Ulrich CM, Potter JD (2006) Folate supplementation: too much of a good thing. Cancer Epidemiol Biomark Prev 15(2):189–193CrossRefGoogle Scholar
  15. 15.
    Laiño JE, del Valle MJ, de Giori GS, LeBlanc JGJ (2014) Applicability of a Lactobacillus amylovorus strain as co-culture for natural folate bio-enrichment of fermented milk. Int J Food Microbiol 191:10–16CrossRefGoogle Scholar
  16. 16.
    Scott JM (1999) Folate and vitamin B 12. Proc Nutr Soc 58(2):441–448CrossRefGoogle Scholar
  17. 17.
    Haghshenas B, Abdullah N, Nami Y, Radiah D, Rosli R, Khosroushahi AY (2014) Different effects of two newly-isolated probiotic Lactobacillus plantarum 15HN and Lactococcus lactis subsp. Lactis 44Lac strains from traditional dairy products on cancer cell lines. Anaerobe 30:51–59CrossRefGoogle Scholar
  18. 18.
    Crittenden R, Martinez N, Playne M (2003) Synthesis and utilisation of folate by yoghurt starter cultures and probiotic bacteria. Int J Food Microbiol 80(3):217–222CrossRefGoogle Scholar
  19. 19.
    Haghshenas B, Nami Y, Almasi A, Abdullah N, Radiah D, Rosli R, Barzegari A, Khosroushahi AY (2017) Isolation and characterization of probiotics from dairies. Iran J Microbiol 9(4):234Google Scholar
  20. 20.
    Krasaekoopt W, Bhandari B, Deeth H (2004) The influence of coating materials on some properties of alginate beads and survivability of microencapsulated probiotic bacteria. Int Dairy J 14(8):737–743CrossRefGoogle Scholar
  21. 21.
    Lin MY, Young CM (2000) Folate levels in cultures of lactic acid bacteria. Int Dairy J 10(5–6):409–413. CrossRefGoogle Scholar
  22. 22.
    Holt DL, Wehling RL, Zeece MG (1988) Determination of native folates in milk and other dairy products by high-performance liquid chromatography. J Chromatogr A 449:271–279CrossRefGoogle Scholar
  23. 23.
    Verwei M, Arkbåge K, Havenaar R, van den Berg H, Witthöft C, Schaafsma G (2003) Folic acid and 5-methyltetrahydrofolate in fortified milk are bioaccessible as determined in a dynamic in vitro gastrointestinal model. J Nutr 133(7):2377–2383CrossRefGoogle Scholar
  24. 24.
    Gangadharan D, Nampoothiri KM (2011) Folate production using Lactococcus lactis ssp cremoris with implications for fortification of skim milk and fruit juices. LWT Food Sci Technol 44(9):1859–1864CrossRefGoogle Scholar
  25. 25.
    Meucci A, Rossetti L, Zago M, Monti L, Giraffa G, Carminati D, Tidona F (2018) Folates biosynthesis by Streptococcus thermophilus during growth in milk. Food Microbiol 69:116–122CrossRefGoogle Scholar
  26. 26.
    Hugenschmidt S, Schwenninger SM, Lacroix C (2011) Concurrent high production of natural folate and vitamin B12 using a co-culture process with Lactobacillus plantarum SM39 and Propionibacterium freudenreichii DF13. Process Biochem 46(5):1063–1070. CrossRefGoogle Scholar
  27. 27.
    Da Silva FFP, Biscola V, LeBlanc JG, de Melo Franco BDG (2016) Effect of indigenous lactic acid bacteria isolated from goat milk and cheeses on folate and riboflavin content of fermented goat milk. LWT Food Sci Technol 71:155–161CrossRefGoogle Scholar
  28. 28.
    Jägerstad M, Jastrebova J, Svensson U (2004) Folates in fermented vegetables, a pilot study. LWT Food Sci Technol 37(6):603–611CrossRefGoogle Scholar
  29. 29.
    Sybesma W, Starrenburg M, Tijsseling L, Hoefnagel MH, Hugenholtz J (2003) Effects of cultivation conditions on folate production by lactic acid bacteria. Appl Environ Microbiol 69(8):4542–4548CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Mohammad Khalili
    • 1
  • Aziz Homayouni Rad
    • 2
  • Ahmad Yari Khosroushahi
    • 3
  • Hadi Khosravi
    • 4
  • Somayeh Jafarzadeh
    • 2
    Email author
  1. 1.Multiple Sclerosis Research Center, Neuroscience InstituteTehran University of Medical SciencesTehranIran
  2. 2.Department of Food Science and Technology, Faculty of Health and NutritionTabriz University of Medical SciencesTabrizIran
  3. 3.Department of Pharmacognosy, Faculty of PharmacyTabriz University of Medical SciencesTabrizIran
  4. 4.Department of Applied Human NutritionMount Saint Vincent UniversityHalifaxCanada

Personalised recommendations