Skip to main content
Log in

Antioxidative potential of folate producing probiotic Lactobacillus helveticus CD6

  • Original Article
  • Published:
Journal of Food Science and Technology Aims and scope Submit manuscript

Abstract

Folate producing Lactobacillus sp. CD6 isolated from fermented milk showed 98% similarity with Lactobacillus helveticus based on 16S rRNA gene sequence analysis. It was found to produce a folic acid derivative 5-methyl tetrahydrofolate (5-MeTHF). The intracellular cell-free extract of strain demonstrated antioxidative activity with the inhibition rate of ascorbate autoxidation in the range of 27.5% ± 3.7%. It showed highest metal ion chelation ability for Fe2+ (0.26 ± 0.06 ppm) as compared to Cu2+. The DPPH (α,α-Diphenyl-β-Picrylhydrazyl) radical scavenging activity for intact cells were found to be 24.7% ± 10.9% proved its antioxidative potential. Furthermore, it demonstrated 14.89% inhibition of epinephrine autoxidation, 20.9 ± 1.8 μg cysteine equivalent reducing activity and 20.8% ± 0.9% hydroxyl radical scavenging effect. The strain was evaluated for probiotic properties as per WHO and FAO guidelines. It showed 90.61% survival at highly acidic condition (pH 2.0), 90.66% viability in presence of synthetic gastric juice and 68% survivability at 0.5% bile concentration for 24 h. It was susceptible to many antibiotics which reduces the prospect to offer resistance determinants to other organisms if administered in the form of probiotic preparations. It showed in vitro mucus binding and antimicrobial activity against enteric pathogens like Salmonella typhimurium (NCIM 2501), Streptococcus pyogenes (NCIM 2608), and Staphylococcus aureus (NCIM 5021) and moreover it showed non- hemolytic activity on sheep blood agar.

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.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Agerholm-Larsen L, Raben A, Haulik N, Hansen AS, Manders M, Astrup A (2000) Effects of 8 weeks intake of probiotic milk products on risk factors for cardiovascular diseases. Eur J Clin Nutr 54:288–297

    Article  CAS  Google Scholar 

  • Ahire JJ, Patil KP, Chaudhari BL, Chincholkar SB (2010) Bacillus spp. of human origin: a potential siderophoregenic probiotic bacteria. Appl Biochem Biotechnol. doi:10.1007/s12010-010-9142-6

    Google Scholar 

  • Ahire JJ, Patil KP, Chaudhari BL, Chincholkar SB (2011) A potential probiotic culture ST2 produces siderophore 2, 3-dihydroxybenzoylserine under intestinal conditions. Food Chem. doi:10.1016/j.foodchem.2010.12.126

    Google Scholar 

  • Alander M, Korpela R, Saxelin M, Vilpponen-Salmela T, Matilla-Sandholm T, Wright A (1997) Recovery of Lactobacillus rhamnosus GG from human colonic biopsies. Lett Appl Microbiol 24:361–364

    Article  CAS  Google Scholar 

  • Anonymous (1990) Probiotics: a review of some of the products currently available to compound feed manufacturers. Feed Compd 5:58–64

    Google Scholar 

  • Axelsson L (2004) Lactic acid bacteria microbiological and functional aspect. In: Salminen S, Wright A, Ouwehand A (eds) (3rd ed.). Marcel & Dekker, New York USA, pp 1–30

  • Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and assay applicable to acrylamide gels. Ana Biochem 44:276–287

    Article  CAS  Google Scholar 

  • Brashears MM, Jaron D, Trimble J (2003) Isolation, selection and characterization of lactic acid bacteria for a competitive exclusion product to reduce shedding of Escherichia coli O157:H7in cattle. J Food Prot 66:355–363

    CAS  Google Scholar 

  • Brosius JM, Palmer L, Kennedy P, Noller HF (1978) Complete nucleotide sequence of the 16S ribosomal DNA gene from Escherichia coli. Proc Natl Acad Sci USA 75:4801–4805

    Article  CAS  Google Scholar 

  • Buettner GR (1990) Use of ascorbate as test for catalytic metals in sample buffers. Methods Enzymol 186:125–127

    Article  CAS  Google Scholar 

  • Collado MC, Meriluoto J, Salminen S (2008) Adhesion and aggregation properties of probiotic and pathogen strains. Eur Food Res Technol 226:1065–1073

    Article  CAS  Google Scholar 

  • Coppola R, Succi M, Tremonte P, Reale A, Salzano G, Sorrentino E (2005) Antibiotic susceptibility of L. rhamnosus strains isolated from Parmigiano Reggiano cheese. Lait 85:193–204

    Article  CAS  Google Scholar 

  • Del Re B, Sgorbati B, Miglioli M, Palenzona D (2000) Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum. Lett Appl Microbiol 31:438–442

    Article  Google Scholar 

  • Diplock AT, Aggett P, Ashwell M, Bornet F, Fern E, Roberfroid M (1999) Scientific concepts of functional foods in Europe: consensus document. Br J Nutr 81:S1–S27

    CAS  Google Scholar 

  • Duhutrel P, Bordat C, Wu TD, Zagorec M, Guerquin-Kern JL, Champomier-Verges MC (2010) Iron sources used by the nonpathogenic lactic acid bacterium Lactobacillus sakei as revealed by electron energy loss spectroscopy and secondary-ion mass spectrometry. Appl Environ Microbiol 76:560–565

    Article  CAS  Google Scholar 

  • Duthie SJ, Narayanan S, Brand GM, Pirie L, Grant G (2002) Impact of folate deficiency on DNA stability. J Nutr 132:2444S–2449S

    CAS  Google Scholar 

  • Evans P, Halliwell B (2001) Micronutrients: oxidant/antioxidant status. Br J Nutr 85(S2):S67–S74

    Article  CAS  Google Scholar 

  • FAO/WHO (2002) Guidelines for the evaluation of probiotics in food, report of a joint FAO/WHO working group on drafting guidelines for the evaluation of probiotics in food, London, Ontario, Canada, pp 1–11

  • Gangadharan D, Sivaramakrishnan S, Pandey A, Nampoothiri MK (2010) Folate-producing lactic acid bacteria from cow’s milk with probiotic characteristics. Int J Dairy Technol 63:339–348

    Article  CAS  Google Scholar 

  • Gardner HW (1975) Decomposition of linoleic acid hydroperoxides. J Agric Food Chem 23:129–136

    Article  CAS  Google Scholar 

  • Ghosh D, Chattopadhyay P (2010) Preparation of idli batter, its properties and nutritional improvement during fermentation. J Food Sci Technol. doi:10.1007/s13197-010-0148-4

    Google Scholar 

  • Gliszczyńska-Świgło A, Muzolf M (2007) pH-dependent radical scavenging activity of folates. J Agric Food Chem 55:8237–8242

    Article  Google Scholar 

  • Grice HC (1988) Safety evaluation of butylated hydroxyanisole from the perspective of effects on forestomach and oesophageal squamous epithelium. J Food Chem Toxicol 26:717–723

    Article  CAS  Google Scholar 

  • Halliwell B, Chirico S (1993) Lipid peroxidation: its mechanism, measurement, and significance. Am J Clin Nutr 57:715S–725S

    CAS  Google Scholar 

  • Halliwell B, Gutteridge JMC (1999) Free radicals in biology and medicine, vol 3. Oxford University Press, New York

    Google Scholar 

  • Holt JH, Kreig NR, Sneath PHA, Staley JT, Williams ST (1994) Bergey’s manual of determinative bacteriology. In: WR Hensyl (ed) (9th ed). Williams & Wilkins Baltimore, MD, USA, pp 1208–1234

  • Jin M, Cai Y, Li J, Zhao H (1996) 1, 10-Phenanthroline Fe2+ oxidative assay of hydroxyl radical produced by H2O2/ Fe2+. Prog Biochem Biophy 23:553–555

    CAS  Google Scholar 

  • Kaizu H, Sasaki M, Nakajima H, Suzuki Y (1993) Effect of antioxidative lactic acid bacteria on rats fed a diet deficient in vitamin E. J Dairy Sci 76:2493–2499

    Article  CAS  Google Scholar 

  • Kuhle A, Skovgaard K, Jespersen L (2005) In vitro screening of probiotic properties of Saccharomyces cerevisae var. boulardii and food borne Saccharomyces cerevisae strains. Int J Food Microbiol 101:29–39

    Article  Google Scholar 

  • Lin MY, Yen CL (1999) Antioxidative ability of Lactic acid bacteria. J Agric Food Chem 47:1460–1466

    Article  CAS  Google Scholar 

  • Looyenga R, Boltz WDF (1971) Spectrophotometric study of the determination of copper with ammonium 1-pyrrolidinecarbodithioate. Talanta 19:82–87

    Google Scholar 

  • Metchnikoff E (1908) Prolongation of life, optimistic studies. Putnam, New York, pp 161–183

    Google Scholar 

  • Mishra OP, Kovachich GB (1984) Inhibition of the autoxidation of ascorbate and norepinephrine by extracts of Clostridium butyrich, Megasphaera elsdenii, and Escherichia coli. Life Sci 35:849–854

    Article  CAS  Google Scholar 

  • Naidu AS, Bidlack WR, Clemens RA (1999) Probiotic spectra of lactic acid bacteria (LAB). Crit Rev Food Sci Nutr 38:13–126

    Article  Google Scholar 

  • Na-kyoung L, Yun CW, Kim SW, Chang HI, Kang CW, Paik HD (2008) Screening of Lactobacilli derived from chicken feces and partial characterization of Lactobacillus acidophilus A12 as animal probiotics. J Microbiol Biotechnol 18:338–342

    Google Scholar 

  • Oyaizu M (1986) Antioxidative activity of browning products of glucosamine fractionated by organic solvent and thin-layer chromatography. Nippon Shokuhin Kogyo Gakksishi 35:771–775

    Article  Google Scholar 

  • Patel AK, Ahire JJ, Pawar S, Chaudhari BL, Shouche YS, Chincholkar SB (2010) Evaluation of the probiotic characteristics of siderophoregenic Bacillus spp. isolated from dairy waste. Appl Biochem Biotechnol 160:140–155

    Article  CAS  Google Scholar 

  • Pedersen C, Lindberg E, Roos S (2004) Microbiological characterization of wet wheat distillers’ grain with focus on isolation of Lactobacilli with potential as probiotics. Appl Environ Microbiol 70:1522–1527

    Article  CAS  Google Scholar 

  • Rezk BM, Haenen GRMM, van der Vijgh WJF, Bast A (2003) Tetrahydrofolate and 5-methyltetrahydrofolate are folates with high antioxidant activity. Identification of the antioxidant pharmacophore. FEBS Lett 555:601–605

    Article  CAS  Google Scholar 

  • Rosenberg M, Gutnick D, Rosenberg E (1980) Adherence of bacteria to hydrocarbons: a simple method for measuring cell-surface hydrophobicity. FEMS Microbiol Lett 9:29–33

    Article  CAS  Google Scholar 

  • Salminen S, Bouley MC, Boutron-Rualt MC, Cummings J, Franck A, Gibson G, Isolauri E, Moreau MC, Roberfroid M, Rowland I (1998) Functional food science and gastrointestinal physiology and function. Bri J Nutr (suppl 1):147–171

  • Sanders JW, Leehouts KJ, Haandrikmam AJ, Venema G, Kok J (1995) Stress response in Lactococcus lactis: cloning, expression analysis and mutation of the lactococcal super oxide dismutase gene. J Bacteriol 177:5254–5260

    CAS  Google Scholar 

  • Sarri JT (1989) Chronic treatment with dimethyl sulphoxide protects against cardiovascular defects of copper deficiency. Proc Soc Exp Biol Med 190:121–124

    Google Scholar 

  • Seifried HE, Anderson DE, Fisher EI, Milner JA (2007) A review of the interaction among dietary antioxidants and reactive oxygen species. J Nutr Biochem 18:567–579

    Article  CAS  Google Scholar 

  • Shane B, Bognar AL, Goldfarb RD, LeBowitz JH (1983) Regulation of folylpolygamma-glutamate synthesis in bacteria: in vivo and in vitro synthesis of pteroylpoly- gammaglutamates by Lactobacillus casei and Streptococcus faecalis. J Bacteriol 153:316–325

    CAS  Google Scholar 

  • Shimada K, Fujikawa K, Yahara K, Nakamura T (1992) Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem 40:945–948

    Article  CAS  Google Scholar 

  • Simic MG (1988) Mechanisms of inhibition of free-radical processed in mutagenesis and carcinogensis. Mutat Res 202:377–386

    Article  CAS  Google Scholar 

  • Smith MA, Harris PLR, Sayre LM, Perry G (1997) Iron accumulation in Alzheimer disease is a source of redox-generated free radicals. Proc Natl Acad Sci USA 94:9866–9868

    Article  CAS  Google Scholar 

  • Stanger O (2002) Physiology of folic acid in health and disease. Curr Drug Metabol 3:211–223

    Article  CAS  Google Scholar 

  • Sun M, Zigman S (1978) An improved spectrophotometric assay for superoxide dismutase based on epinephrine autoxidation. Anal Biochem 90:81–89

    Article  CAS  Google Scholar 

  • Vakil JR, Shahani KM (1968) Disc assay method for determination of folic acid content of milk, cheese, and other foods. J Dairy Sci 52:325–328

    Article  Google Scholar 

  • Valko M, Leibfritz D, Moncol J, Cronin MTD, Mazur M, Telser J (2007) Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 39:44–84

    Article  CAS  Google Scholar 

  • Vanaja G, Gotcheva V, Angelov A, Agrawal R (2011) Formation of volatiles and fattyacids of therapeutic importance in the probiotic Lactobacillus plantarum LPcfr adapted to resist GIT conditions. J Food Sci Technol 48:110–113

    Article  CAS  Google Scholar 

  • Verma AK, Banerjee R (2010) Dietary fibre as functional ingredient in meat products: a novel approach for healthy living—a review. J Food Sci Technol 47:247–257

    Article  Google Scholar 

  • Wang AN, Yi XW, Yu HF, Dong B, Qiao SY (2009) Free radical scavenging activity of Lactobacillus fermentum in vitro and its antioxidative effect on growing–finishing pigs. Appl Microbiol 107:1140–1148

    Article  CAS  Google Scholar 

  • Wolff T, Witkop TC, Miller T, Syed BS (2009) Folic acid supplementation for the prevention of neural tube defects: an update of the evidence for the U.S. preventive services task force. Ann Intern Med 150:632–639

    Google Scholar 

  • Yamauchi R, Goto Y, Kato K, Ueno Y (1984) Prooxidant effect on dihydroxyacetone and reducing sugars on the autoxidation of methyl emulsions. J Agric Biol Chem 48:843–848

    Article  CAS  Google Scholar 

Download references

Acknowledgement

The authors are thankful to University Grants Commission, New Delhi for financial support.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Bhushan Liladhar Chaudhari.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ahire, J.J., Mokashe, N.U., Patil, H.J. et al. Antioxidative potential of folate producing probiotic Lactobacillus helveticus CD6. J Food Sci Technol 50, 26–34 (2013). https://doi.org/10.1007/s13197-011-0244-0

Download citation

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13197-011-0244-0

Keywords

Navigation