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Lactobacillus intestinalis efficiently produces equol from daidzein and chungkookjang, short-term fermented soybeans

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Abstract

Equol improves menopausal symptoms and it is synthesized from daidzein, one of the isoflavonoids in soybeans, by the bacteria in the large intestines of some people. The purpose of this study was to isolate equol-producing bacteria using daidzein from the intestinal microflora and to produce equol-containing chungkookjang (short-term fermented soybean). Equol-producing bacteria from the feces of Sprague–Dawley female rats were isolated using media containing daidzein. The isolated bacteria were cultured in thioglycollate media and equol production was identified through thin-layer chromatography and ultraperformance liquid chromatography–mass spectrometry. The bacteria were identified by 16S rRNA sequencing. The rate of equol production in different concentrations of daidzein was assessed. The expression of genes that code for enzymes associated with the production of equol from daidzein was detected through reverse transcription quantitative PCR. The bacterium we isolated was Lactobacillus intestinalis (LC096206.1, 99%). L. intestinalis was found to express daidzein reductase, dihydrodaidzein reductase, and tetrahydrodaidzein reductase, the enzymes involved in producing equol from daidzein. The conversion rate of equol from daidzein was highest (29.5%) using 200 μM daidzein for 48 h of incubation. When chungkookjang fermented with Bacillus amyloquencies SRCM100001 was incubated with L. intestinalis, 0.32 ± 0.04 mg equol/g chungkookjang was produced. In conclusion, L. intestinalis efficiently produces equol from not only daidzein but also in chungkookjang.

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References

  • Abiru Y, Kumemura M, Ueno T, Uchiyama S, Masaki K (2012) Discovery of an S-equol rich food stinky tofu, a traditional fermented soy product in Taiwan. Int J Food Sci Nutr 63:964–970

    Article  CAS  PubMed  Google Scholar 

  • Ahmadi SM, Khomiri M, Khosroushahi A, Kashaninezhad M (2009) Isolation and identification of lactic acid bacteria (LAB) from Iranian traditional Lighvan cheese. J Agric Sci Nat Res 16:136–146

    Google Scholar 

  • Akaza H et al (2004) Comparisons of percent equol producers between prostate cancer patients and controls: case-controlled studies of isoflavones in Japanese, Korean and American residents. Jpn J Clin Oncol 34:86–89

    Article  PubMed  Google Scholar 

  • An-yi W, Yong-qiang C, Xiao-qun O, Xiao-nan L, Ting L (2014) Thin-layer chromatography (TLC) high performance liquid chromatography(HPLC) equol genistein. Sci Technol Food Ind 35:79–82

    Google Scholar 

  • Atkinson C, Frankenfeld CL, Lampe JW (2005) Gut bacterial metabolism of the soy isoflavone daidzein: exploring the relevance to human health. Exp Biol Med 230:155–170

    Article  CAS  Google Scholar 

  • Axelson M, Kirk DN, Farrant RD, Cooley G, Lawson AM, Setchell KD (1982) The identification of the weak oestrogen equol [7-hydroxy-3-(4′-hydroxyphenyl)chroman] in human urine. Biochem J 201:353–357

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen MN, Lin CC, Liu CF (2015) Efficacy of phytoestrogens for menopausal symptoms: a meta-analysis and systematic review. Climacteric 18:260–269

    Article  CAS  PubMed  Google Scholar 

  • Clarkson TB et al (2011) The role of soy isoflavones in menopausal health: report of The North American Menopause Society/wulf H. Utian Translational Science Symposium in Chicago, Il (October 2010). Menopause 18:732–753

    Article  Google Scholar 

  • Daily JW, Ko BS, Ryuk J, Liu M, Zhang W, Park S (2019) Equol decreases hot flashes in postmenopausal women: a systematic review and meta-analysis of randomized clinical trials. J Med Food 22:127–139

    Article  PubMed  Google Scholar 

  • Decroos K, Vanhemmens S, Cattoir S, Boon N, Verstraete W (2005) Isolation and characterisation of an equol-producing mixed microbial culture from a human faecal sample and its activity under gastrointestinal conditions. Arch Microbiol 183:45–55

    Article  CAS  PubMed  Google Scholar 

  • Frankenfeld CL et al (2005) High concordance of daidzein-metabolizing phenotypes in individuals measured 1 to 3 years apart. Br J Nutr 94:873–876

    Article  CAS  PubMed  Google Scholar 

  • Fujisawa T, Itoh K, Benno Y, Mitsuoka T (1990) Lactobacillus intestinalis (ex Hemme 1974) sp. nov., nom. rev., isolated from the intestines of mice and rats. Int J Syst Bacteriol 40:302–304

    Article  CAS  PubMed  Google Scholar 

  • Guadamuro L, Dohrmann AB, Tebbe CC, Mayo B, Delgado S (2017) Bacterial communities and metabolic activity of faecal cultures from equol producer and non-producer menopausal women under treatment with soy isoflavones. BMC Microbiol 17:93

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hur HG, Lay JO Jr, Beger RD, Freeman JP, Rafii F (2000) Isolation of human intestinal bacteria metabolizing the natural isoflavone glycosides daidzin and genistin. Arch Microbiol 174:422–428

    Article  CAS  PubMed  Google Scholar 

  • Jackson RL, Greiwe JS, Schwen RJ (2011) Emerging evidence of the health benefits of S-equol, an estrogen receptor beta agonist. Nutr Rev 69:432–448

    Article  PubMed  Google Scholar 

  • Kwon DY et al (2007) Long-term consumption of fermented soybean-derived Chungkookjang enhances insulinotropic action unlike soybeans in 90% pancreatectomized diabetic rats. Eur J Nutr 46:44–52

    Article  CAS  PubMed  Google Scholar 

  • Landete JM, Arques J, Medina M, Gaya P, de Las Rivas B, Munoz R (2016) Bioactivation of phytoestrogens: intestinal bacteria and health. Crit Rev Food Sci Nutr 56:1826–1843

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Matthies A, Loh G, Blaut M, Braune A (2011) Daidzein and genistein are converted to equol and 5-hydroxy-equol by human intestinal Slackia isoflavoniconvertens in gnotobiotic rats–3. J Nutr 142:40–46

    Article  CAS  PubMed  Google Scholar 

  • Nagata C, Shimizu H, Takami R, Hayashi M, Takeda N, Yasuda K (1999) Hot flushes and other menopausal symptoms in relation to soy product intake in Japanese women. Climacteric 2:6–12

    Article  CAS  PubMed  Google Scholar 

  • Newton KM, Reed SD, LaCroix AZ, Grothaus LC, Ehrlich K, Guiltinan J (2006) Treatment of vasomotor symptoms of menopause with black cohosh, multibotanicals, soy, hormone therapy, or placebo: a randomized trial. Ann Intern Med 145:869–879

    Article  PubMed  Google Scholar 

  • Nishimura Y et al (2017) S-equol exerts estradiol-like anorectic action with minimal stimulation of estrogen receptor-alpha in ovariectomized rats. Front Endocrinol (Lausanne) 8:281

    Article  Google Scholar 

  • Saitoh S, Sato T, Harada H, Matsuda T (2004) Biotransformation of soy isoflavone-glycosides in laying hens: intestinal absorption and preferential accumulation into egg yolk of equol, a more estrogenic metabolite of daidzein. Biochim Biophys Acta 1674:122–130

    CAS  PubMed  Google Scholar 

  • Schroder C, Matthies A, Engst W, Blaut M, Braune A (2013) Identification and expression of genes involved in the conversion of daidzein and genistein by the equol-forming bacterium Slackia isoflavoniconvertens. Appl Environ Microbiol 79:3494–3502

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Setchell KDR, Clerici C (2010) Equol: history, chemistry, and formation, 2. J Nutr 140:1355S–1362S

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Setchell KD, Cole SJ (2006) Method of defining equol-producer status and its frequency among vegetarians. J Nutr 136:2188–2193

    Article  CAS  PubMed  Google Scholar 

  • Setchell KD, Brown NM, Lydeking-Olsen E (2002) The clinical importance of the metabolite equol-a clue to the effectiveness of soy and its isoflavones. J Nutr 132:3577–3584

    Article  CAS  PubMed  Google Scholar 

  • Tamura M, Tsushida T, Shinohara K (2007) Isolation of an isoflavone-metabolizing, Clostridium-like bacterium, strain TM-40, from human faeces. Anaerobe 13:32–35

    Article  CAS  PubMed  Google Scholar 

  • Tsen SY, Tan XY, Tan YM, Yan BY, Loke WM (2016) Relative inhibitions of 5-lipoxygenase and myeloperoxidase and free-radical scavenging activities of daidzein, dihydrodaidzein, and equol. J Med Food 19:543–548

    Article  CAS  PubMed  Google Scholar 

  • Vazquez L, Guadamuro L, Giganto F, Mayo B, Florez AB (2017) Development and use of a Real-Time Quantitative PCR Method for detecting and quantifying Equol-Producing bacteria in human faecal samples and slurry cultures. Front Microbiol 8:1155

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang XL, Hur HG, Lee JH, Kim KT, Kim SI (2005) Enantioselective synthesis of S-equol from dihydrodaidzein by a newly isolated anaerobic human intestinal bacterium. Appl Environ Microbiol 71:214–219

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang XL, Kim HJ, Kang SI, Kim SI, Hur HG (2007) Production of phytoestrogen S-equol from daidzein in mixed culture of two anaerobic bacteria. Arch Microbiol 187:155–160

    Article  CAS  PubMed  Google Scholar 

  • Woo PC, Leung PK, Leung KW, Yuen KY (2000) Identification by 16S ribosomal RNA gene sequencing of an Enterobacteriaceae species from a bone marrow transplant recipient. Mol Pathol 53:211–215

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xiao-mei L, Meng-han J, Yao J (2017) An assessment on the individual & collective equol cultivating productivity of intestinal bacteria. Sci Technol Food Ind 8:281–291

    Google Scholar 

  • Xue J, Hong-yu L (2014) Optimization of fermentation medium components for equol production using response surface methodology. Food Ind 35:67–71

    Google Scholar 

  • Yang HJ et al (2013) Soybean fermentation with Bacillus licheniformis increases insulin sensitizing and insulinotropic activity. Food Funct 4:1675–1684

    Article  CAS  PubMed  Google Scholar 

  • Yokoyama S, Suzuki T (2008) Isolation and characterization of a novel equol-producing bacterium from human feces. Biosci Biotechnol Biochem 72:2660–2666

    Article  CAS  PubMed  Google Scholar 

  • Yu ZT, Yao W, Zhu WY (2008) Isolation and identification of equol-producing bacterial strains from cultures of pig faeces. FEMS Microbiol Lett 282:73–80

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This work was supported by the Main Research Program (E0150302) of the Korea Food Research Institute (KFRI) and the Ministry of Science, ICT & Future Planning and Traditional Culture Convergence Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (NRF- 2016M3C1B5907152).

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Author notes

  1. Yuan Heng and Min Jung Kim contributed equally to this work.

Authors and Affiliations

  1. Department of Food and Nutrition, Obesity/Diabetes Center, Hoseo University, 165 Sechul-Ri, BaeBang-Yup, Asan-Si, Chungnam-Do, 336-795, South Korea

    Yuan Heng, Suna Kang & Sunmin Park

  2. Food Functional Research Division, Korean Food Research Institutes, Sungnam, South Korea

    Min Jung Kim & Hye Jeong Yang

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  1. Yuan Heng
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  2. Min Jung Kim
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  4. Suna Kang
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Contributions

Conceived and designed the experiments: SP. Performed the experiments: YH, MJK. Analyzed the data: SK, YH, MJK, HJY. Wrote the draft paper: SP, YH, SK, JYZ. All authors read and approved the final manuscript.

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Correspondence to Sunmin Park.

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Communicated by Erko Stackebrandt.

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Heng, Y., Kim, M.J., Yang, H.J. et al. Lactobacillus intestinalis efficiently produces equol from daidzein and chungkookjang, short-term fermented soybeans. Arch Microbiol 201, 1009–1017 (2019). https://doi.org/10.1007/s00203-019-01665-5

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  • DOI: https://doi.org/10.1007/s00203-019-01665-5

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