Skip to content
BY-NC-ND 3.0 license Open Access Published by De Gruyter Open Access March 21, 2015

Proteomic proof that a probiotic elevates glutathione level in human serum

  • Aune Rehema , Tiiu Kullisaar , Kärt Seer , Kristen Reinmann , Mihkel Zilmer and Kalle Kilk
From the journal Open Life Sciences


Lactobacillus fermentum ME-3 (LfME-3) has been proven to synthesize and secrete glutathione. A regular use of the foods fermented by it has shown a favourable influence on human lipid profiles and several antioxidant parameters. We administered the LfME-3-fermented kefir for 14 days to 43 human subjects and evaluated their serum with MALDI-TOF mass spectrometer at the beginning and end of the test period. We found an increase of the peak at m/z 308 (corresponding to glutathione) and a new peak at m/z 1467.


[1] Lin M.Y., Chang F.Y., Antioxidative effect of intestinal bacteria Bifidobacterium longum ATCC15708 and Lactobacillus acidophilus ATCC 4356, Dig. Dis. Sci., 2000, 45, 1617–1622 Search in Google Scholar

[2] McFarland L.V., Beneficial microbes: health or hazard?, Eur. J. Gastroenterol. Hepatol., 2000, 12, 1069–1071 10.1097/00042737-200012100-00001Search in Google Scholar

[3] Isolauri E . Probiotics in human disease, Am. J. Clin. Nutr., 2001, 73, 1142S–1146S 10.1093/ajcn/73.6.1142SSearch in Google Scholar

[4] Kullisaar T., Songisepp E., Mikelsaar M., Zilmer K., Vihalemm, T., Zilmer, M., Antioxidative probiotic goats’ milk decreases oxidative stress mediated atherogenicity in human subjects, Br. J. Nutr., 2003, 90, 449-456 10.1079/BJN2003896Search in Google Scholar

[5] Kullisaar T., Shepetova J., Zilmer K.; Songisepp E., Rehema, A., Mikelsaar M., et al., An antioxidant probiotic reduces postprandial lipemia and oxidative stress, Centr. Eur. J. Biol., 2011, 6, 32–40 10.2478/s11535-010-0103-4Search in Google Scholar

[6] Sepp E., Julge K., Vasar M., Naaber P., Björgsten B., Mikelsaar M., Intestinal microflora of Estonian and Swedish children, Acta Pediatr., 1997, 86, 956–961 10.1111/j.1651-2227.1997.tb15178.xSearch in Google Scholar

[7] Mikelsaar M., Annuk H., Shchepetova J., Mändar R., Sepp E., Björksten B., Intestinal lactobacilli of Estonian and Swedish children, Microb. Ecol. Health Dis., 2002, 14, 75–80 10.1080/08910600260081711Search in Google Scholar

[8] Kullisaar T., Zilmer M., Mikelsaar M., Vihalemm T., Annuk H., Kairane C, et al., Two antioxidative lactobacilli strains as promising probiotics, Int. J. Food Microbiol., 2002, 72, 215–224 10.1016/S0168-1605(01)00674-2Search in Google Scholar

[9] Mikelsaar M., Zilmer M., Lactobacillus fermentum ME-3 - an antimicrobial and antioxidative probiotic. Microb. Ecol. Health Dis., 2009, 21, 1-27 10.1080/08910600902815561Search in Google Scholar PubMed PubMed Central

[10] Kullisaar T., Songisepp E., Aunapuu M., Kilk K., Arend A., Mikelsaar, M., et al., Complete Glutathione System in Probiotic Lactobacillus fermentum ME-3, Appl. Biochem. Microbiol. (Pleiades Publishing), 2010, 46, 481–486 10.1134/S0003683810050030Search in Google Scholar

[11] Yoon Y.H., Byun J.R., Occurrence of Glutathione Sulphydryl (GSH) and Antioxidant Activities in Probiotic Lactobacillus spp., Asian-Aust. J. Anim .Sci., 2004, 17, 1582–1585 10.5713/ajas.2004.1582Search in Google Scholar

[12] Peran L., Camuesco D., Comalada M., Bailon E., Henriksson A., Xaus J., et al., A comparative study of the preventative effects exerted by three probiotics, Bifidobacterium lactis, Lactobacillus casei and Lactobacillus acidophilus, in the TNBS model of rat colitis, J. Appl. Microbiol., 2007, 103, 836–844 10.1111/j.1365-2672.2007.03302.xSearch in Google Scholar PubMed

[13] Lee K., Pi K., Kim E.B., Rho B.S., Kang S.K., Lee H.G., et al., Glutathione-mediated response to acid stress in the probiotic bacterium, Lactobacillus salivarius, Biotechnol. Lett., 2010 , 32, 969-72 10.1007/s10529-010-0244-6Search in Google Scholar PubMed

[14] Monostori P., Wittmann G., Karg E,; Tửri S., Determination of glutathione and glutathione disulfide in biological samples: an in-depth review, J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 2009, 877, 3331-3346 10.1016/j.jchromb.2009.06.016Search in Google Scholar PubMed

[15] Zilmer M., Soomets U., Rehema A., Langel Ü., The glutathione system as an attractive therapeutic target, Drug Design Reviews., 2005, 2, 121–127 10.2174/1567269053202697Search in Google Scholar

[16] Annuk H., Schepetova J., Kullisaar T., Songisepp E., Zilmer M., Mikelsaar M., Characterization of intestinal lactobacilli as putative probiotic candidates, J. Appl. Microbiol., 2003, 94, 403-412 10.1046/j.1365-2672.2003.01847.xSearch in Google Scholar PubMed

[17] Hamon E., Horvatovich P., Izquierdo E., Bringel F., Marchioni E., Aoudé-Werner D., et al., Comparative proteomic analysis of Lactobacillus plantarum for the identification of key proteins in bile toleranc, BMC Microbiol., 2011, 11, 63 10.1186/1471-2180-11-63Search in Google Scholar PubMed PubMed Central

[18] Kaur S., Kullisaar T., Mikelsaar M., Eisen M., Rehema A., Vihalemm T., et al., Successful management of mild atopic dermatitis in adults with probiotics and emollient, Cent. Eur. J. Med., 2008, 3, 215-220 10.2478/s11536-008-0022-1Search in Google Scholar

[19] Songisepp E., Kals J., Kullisaar T., Mändar R., Hütt P., Zilmer M., et al., Evaluation of the functional efficacy of an antioxidative probiotic in healthy volunteers, Nutr.J., 2005, 4, 22 10.1186/1475-2891-4-22Search in Google Scholar PubMed PubMed Central

[20] Hopps E., Noto D., Caimi G., Averna M.R., A novel component of the metabolic syndrome: The oxidative stress, Nutr. Metab. Cardiovasc. Dis., 2010, 20, 72-77 10.1016/j.numecd.2009.06.002Search in Google Scholar PubMed

[21] Fisher-Wellman K., Bloomer R.J., Macronutrientspecific postprandial oxidative stress: relevance to the development of insulin resistance, Curr. Diabetes Rev., 2009, 5, 228-238 10.2174/157339909789804369Search in Google Scholar PubMed

[22] Witschi A., Reddy S., Stofer B., Lauterburg B.H., The systemic availability of oral glutathione, Eur. J. Clin. Pharmacol., 1992, 43, 667-669 10.1007/BF02284971Search in Google Scholar PubMed

[23] Abeijón Mukdsi M.C, Gauffin Cano M.P., González S.N., Medina R.B., Administration of Lactobacillus fermentum CRL1446 increases intestinal feruloyl esterase activity in mice, Lett. Appl. Microbiol., 2012, 54, 18-25 10.1111/j.1472-765X.2011.03166.xSearch in Google Scholar PubMed

[24] Li Y., Wei G., Chen J., Glutathione: a review on biotechnological production, Appl. Microbiol. Biotechnol., 2004, 66, 233–242 10.1007/s00253-004-1751-ySearch in Google Scholar PubMed

[25] Teunissen C., Koel-Simmelink M., Pham T., Knol J., Khalil M., Trentini A., et al., Identification of biomarkers for diagnosis and progression of MS by MALDI-TOF mass spectrometry, Mult. Scler., 2011, 7, 838-850 10.1177/1352458511399614Search in Google Scholar PubMed

[26] Mateos J., Lourido L., Fernándes-Puente P., Calamia, V., Fernándes-López C., Oreiro N., et al., Differential protein profiling of synovial fluid from rheumatoid arthritis and osteoarthritis patients using LC-MALDI TOF/TOF, J. Proteomics, 2012, 75, 2869-2878 10.1016/j.jprot.2011.12.042Search in Google Scholar PubMed

[27] Feng C.-H., Huang H-Y., Lu C-Y., Quantitation of the glutathione in human peripheral blood by matrix-assisted laser desorption time-of-flight mass spectrometry coupled with micro-scale derivatization, Anal. Chim. A., 2011, 690, 209-214 10.1016/j.aca.2011.02.015Search in Google Scholar PubMed

[28] Kleerebezem M., Hols P., Bernard E., Rolain T., Zhou M., Siezen R.J., et al., The extracellular biology of the lactobacilli, FEMS Microbiol. Rev., 2010, 34, 199-230 10.1111/j.1574-6976.2009.00208.xSearch in Google Scholar

[29] Chertov O., Biragyn A., Kwak L.W., Simpson J.T., Boronina T., Hoang V.M., et al., Organic solvent extraction of proteins and peptides from serum as an effective sample preparation for detection and identification of biomarkers by mass spectrometry. Proteomics, 2004 ,4, 1195-1203 10.1002/pmic.200300677Search in Google Scholar PubMed

[30] Xiang Y., Matsui T., Matsuo K,; Shimada K., Tohma S., Nakamura H., et al., Comprehensive Investigation of Disease- Specific Short Peptides in Sera From Patients With Systemic Sclerosis, Arthritis Rheum., 2007, 56, 2018–2030 10.1002/art.22645Search in Google Scholar PubMed

[31] Che F.Y., Lim J., Pan H., Biswas R., Fricker L.D., Quantitative neuropeptidomics of microwave-irradiated mouse brain and pituitary, Mol. Cell. Proteomics, 2005, 4, 1391-1405 10.1074/mcp.T500010-MCP200Search in Google Scholar PubMed

[32] Chadeau-Hyam M., Hoggart C.J., O´Reilly P.F., Whittaker J.C., De Iorio M., Balding D.J. Fregene: simulation of realistic sequencelevel data in populations and ascertained samples, BMC Bioinformatics 2008, 9, 364 10.1186/1471-2105-9-364Search in Google Scholar

[33] Villanueva J., Martorella A.J., Lawlor K., Philip, J., Fleisher M., Robbins R.J., et al., Serum peptidome patterns that distinguish metastatic thyroid carcinoma from cancer-free controls are unbiased by gender and age, Mol. Cell. Proteomics, 2006, 5, 1840-1852 10.1074/mcp.M600229-MCP200Search in Google Scholar

[34] Vogeser M., Kirchhof, F., Progress in automation of LC-MS in laboratory medicine, Clin. Biochem., 2011, 44, 4–13 10.1016/j.clinbiochem.2010.06.005Search in Google Scholar

[35] Di Girolamo F., Alessandroni J., Somm, P., Guadaqni F., Pre-analytical operating procedures for serum Low Molecular Weight protein profiling, Proteomics, 2010, 73, 667-677 10.1016/j.jprot.2009.09.006Search in Google Scholar

[36] Luque-Garcia J.L., Neubert T.A., Sample preparation for serum/plasma profiling and biomarker identification by mass spectrometry, J. Chromatogr. A., 2007, 1153, 259-276 10.1016/j.chroma.2006.11.054Search in Google Scholar

[37] Kat H., Okamoto M., Yamashita K., Nakamura Y., Fukumori Y., Nakai K., et al., J. Toxicol. Sci., 2003, 28, 19-24 10.2131/jts.28.19Search in Google Scholar

[38] Iwasaki Y., Saito Y., Nakano Y., Mochizuki, K., Sakata, O., Ito, R., et al., Chromatographic and mass spectrometric analysis of glutathione in biological samples, J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci. 2009, 877, 3309-3317 10.1016/j.jchromb.2009.07.001Search in Google Scholar

[39] Gianazza E., Mainini V., Castoldi G., Chinello C., Zerbini G., Bianchi C., et al., Different expression of fibrinopeptide A and related fragments in serum of type 1 diabetic patients with nephropathy, J. Proteomics, 2010, 73, 593-601 10.1016/j.jprot.2009.07.006Search in Google Scholar

[40] Alexandrov T., Decker J., Mertens B., Deelder A.M., Tollenaar R.A., Maass P., et al., Biomarker discovery in MALDI-TOF serum protein profiles using discrete wavelet transformation, Bioinformatics, 2009, 25, 643–649 10.1093/bioinformatics/btn662Search in Google Scholar

[41] Perkins D.N., Pappin D.J.C., Creasy D.M., Cottrell J.S., Probability-based protein identification by searching sequence databases using mass spectrometry data, Electrophoresis, 1999, 20, 3551-3567 10.1002/(SICI)1522-2683(19991201)20:18<3551::AID-ELPS3551>3.0.CO;2-2Search in Google Scholar

[42] Melis G.B., Fruzzetti F., Paoletti, A.M., Carmassi F., Fioretti P., Fibrinopeptide A plasma levels during low-estrogen oral contraceptive treatment, Contraception, 1984, 30, 575-583 10.1016/0010-7824(84)90007-6Search in Google Scholar

[43] Banfi G., Del Fabbro M., Biological variation in tests of hemostasis, Semin. Thromb. Hemost., 2009, 35, 119-126 10.1055/s-0029-1214155Search in Google Scholar PubMed

[44] Allen J., Bradley R.D., Effects of Oral Glutathione Supplementation on Systemic Oxidative Stress Biomarkers in Human Volunteers, J. Altern. Complement. Med., 2011, 17, 827-833 10.1089/acm.2010.0716Search in Google Scholar PubMed PubMed Central

[45] Circu M.L., Aw T.Y., Redox biology of the intestin, Free. Radic. Res., 2011, 45, 1245-1266 10.3109/10715762.2011.611509Search in Google Scholar PubMed PubMed Central

[46] Kern J.K., Geier D.A., Adams J.B., Garver C.R., Audhya T., Geier M.R., A clinical trial of glutathione supplementation in autism spectrum disorders, Med. Sci. Monit., 2011, 17, CR677-682 10.12659/MSM.882125Search in Google Scholar PubMed PubMed Central

[47] Koo S.H., Lee J.S., Kim G.H., Lee H.G., Preparation, characteristics, and stability of glutathione-loaded nanoparticles, J. Agric. Food Chem., 2011, 59, 11264-11269 10.1021/jf2024648Search in Google Scholar PubMed

[48] Peran L., Camuesco D., Comalada M., Nieto A., Concha A., Adrio J.L., et al., Lactobacillus fermentum, a probiotic capable to release glutathione, prevents colonic inflammation in the TNBS model of rat colitis. Int. J. Colorectal Dis., 2006, 21, 737-746 10.1007/s00384-005-0773-ySearch in Google Scholar PubMed

[49] Li Y., Hugenholtz J., Sybesma W., Abee T., Molenaar D. Using Lactococcus lactis for glutathione overproduction, Appl. Microbiol. Biotechnol., 2005, 67, 83-90 10.1007/s00253-004-1762-8Search in Google Scholar PubMed

Received: 2013-10-15
Accepted: 2014-5-26
Published Online: 2015-3-21

©2015 Aune Rehema et al.

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Downloaded on 10.12.2023 from
Scroll to top button