Skip to main content

Advertisement

SpringerLink
Log in

Evaluation of two Bacillus subtilis strains isolated from Korean fermented food as probiotics against loperamide-induced constipation in mice

  • Article
  • Published:
Journal of the Korean Society for Applied Biological Chemistry Submit manuscript

Abstract

Probiotics are live microbes that confer health benefits on the host when administered in adequate amounts. To evaluate the probiotic potential of Bacillus subtilis isolated from Korean fermented foods, we investigated the resistance to biological barriers and improvement of loperamide-induced constipation. The values of resistance to gastric acidity of B. subtilis CBD2 and KMKW4 strains were 55.34±2.12 and 64.58±2.95%, respectively, whereas the survival rate of B. subtilis KMKW4 (31.17±5.78%) in bile acids was superior to that of CBD2 (8.62±2.09%). These strains also demonstrated adhesiveness to intestinal epithelial HT-29 cells and an inhibitory activity against pathogenic microflora. Furthermore, B. subtilis CBD2 and KMKW4 strains improved gastrointestinal activity when tested in a loperamide-induced mouse model of constipation. Pre-treatment with CBD2 and KMKW4 strains before the onset of constipation improved fecal output and gastrointestinal transit in loperamide-treated mice. These strains also showed inhibitory effects on the activity of β-glucosidase and tryptophanase, harmful enzymes of intestinal microflora. Taken together, these finding show that B. subtilis CBD2 and KMKW4 have high adaptability to gastrointestinal environment, and the ability to inhibit pathogenic microflora and prevent constipation, suggesting their activity as potential probiotics.

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

Similar content being viewed by others

References

  • An HM, Baek EH, Jang S, Lee DK, Kim MJ, Kim JR, Lee KO, Park JG, and Ha NJ (2010) Efficacy of Lactic Acid Bacteria (LAB) supplement in management of constipation among nursing home residents. Nutr J 9, 5.

    Article  Google Scholar 

  • An HM, Park SY, Lee DK, Kim JR, Cha MK, Lee SW, Lim HT, Kim KJ, and Ha NJ (2011) Antiobesity and lipid-lowering effects of Bifidobacterium spp. in high fat diet-induced obese rats. Lipids Health Dis 10, 116.

    Article  Google Scholar 

  • Awouters F, Niemegeers CJ, and Janssen PA (1983) Pharmacology of antidiarrheal drugs. Ann Rev Pharmacol Toxicol 23, 279–301.

    Article  CAS  Google Scholar 

  • Begley M, Gahan CG, and Hill C (2005) The interaction between bacteria and bile. FEMS Microbiol Rev 29, 625–51.

    Article  CAS  Google Scholar 

  • Bezkorovainy A (2001) Probiotics: determinants of survival and growth in the gut. Am J Clin Nutr 73, 399S–405S.

    CAS  Google Scholar 

  • Bharucha AE (2007) Constipation. Best Pract Res Clin Gastroenterol 21, 709–31.

    Article  Google Scholar 

  • Blum S, Reniero R, Schiffrin EJ, Crittenden R, Mattila-Sandholm T, Ouwehand AC, Salminen S, Wright A von, Saarela S, Saxelin M, Collins K, and Morelli L (1999) Adhesion studies for probiotics: need for validation and refinement. Trends Food Sci Technol 10, 405–10.

    Article  CAS  Google Scholar 

  • Casula G and Cutting SM (2002) Bacillus probiotics: spore germination in the gastrointestinal tract. App Environ Microbiol 68, 2344–52.

    Article  CAS  Google Scholar 

  • Chadwick RW, George SE, and Claxton LD (1992) Role of the gastrointestinal mucosa and microflora in the bioactivation of dietary and environmental mutagens or carcinogens. Drug Metab Rev 24, 425–92.

    Article  CAS  Google Scholar 

  • Chung KT, Fulk GE, and Slein MW (1975) Tryptophanase of fecal flora as a possible factor in the etiology of colon cancer. J Natl Cancer Inst 54, 1073–8.

    CAS  Google Scholar 

  • Crittenden R, Karppinen S, Ojanen S, Tenkanen M, Fagerström R, Mättö J, Saarela M, Mattila-Sandholm T, and Poutanen K (2002) In vitro fermentation of cereal dietary fibre carbohydrates by probiotic and intestinal bacteria. J Sci Food Agric 82, 781–9.

    Article  CAS  Google Scholar 

  • Cutting SM (2011) Bacillus probiotics. Food Microbiol 28, 214–20.

    Article  Google Scholar 

  • Devries MP, Vessalo M, and Galligan JJ (2010) Deletion of P2X2 and P2X3 receptor subunits does not alter motility of the mouse colon. Front Neurosci 4, 22.

    CAS  Google Scholar 

  • Di Bosco AM, Grieco P, Diurno MV, Campiglia P, Novellino E, and Mazzoni O (2008) Binding site of loperamide: automated docking of loperamide in human mu- and delta-opioid receptors. Chem Biol Drug Des 71, 328–35.

    Article  Google Scholar 

  • Emmanuel AV, Tack J, Quigley EM, and Talley NJ (2009) Pharmacological management of constipation. Neurogastroenterol Motil 21, 41–54.

    Article  CAS  Google Scholar 

  • Erasto P, Adebola PO, Grierison DS, and Afolayan AJ (2005) An ethanobotanical study of plants used for the treatment of diabetes in Eastern Cape Province, South Africa. Afr J Biotechnol 4, 1458–60.

    Google Scholar 

  • Erkkilä S and Petäjä E (2000) Screening of commercial meat starter cultures at low pH and in the presence of bile salts for potential probiotic use. Meat Sci 55, 297–300.

    Article  Google Scholar 

  • Griffiths EA, Duffy LC, Schanbacher FL, Qiao H, Dryja D, and Leavens A (2004) In vivo effects of bifidobacteria and lactoferrin on gut endotoxin concentration and mucosal immunity in Balb/c mice. Dig Dis Sci 49, 579–89.

    Article  CAS  Google Scholar 

  • Guan L, Cho KH, and Lee JH (2011) Analysis of the cultivable bacterial community in jeotgal, a Korean salted and fermented seafood, and identification of its dominant bacteria. Food Microbiol 28, 101–13.

    Article  CAS  Google Scholar 

  • Gudiel-Urbano M and Goñi I (2002) Effect of short-chain fructooligosaccharides and cellulose on cecal enzyme activities in rats. Ann Nutr Metab 46, 254–8.

    Article  CAS  Google Scholar 

  • Guerin-Danan C, Chabanet C, Pedone C, Popot F, Vaissade P, Bouley C, Szylit O, and Andrieux C (1998) Milk fermented with yogurt cultures and Lactobacillus casei compared with yogurt and gelled milk: influence on intestinal microflora in healthy infants. Am J Clin Nutr 67, 111–7.

    CAS  Google Scholar 

  • Heller KJ (2001) Probiotic bacteria in fermented foods: product characteristics and starter organisms. Am J Clin Nutr 73, 374S–9S.

    CAS  Google Scholar 

  • Holzer P (2009) Opioid receptors in the gastrointestinal tract. Regul Pept 155, 11–7.

    Article  CAS  Google Scholar 

  • Hong HA, Huang JM, Khaneja R, Hiep LV, Urdaci MC, and Cutting SM (2008) The safety of Bacillus subtilis and Bacillus indicus as food probiotics. J Appl Microbiol 105, 510–20.

    Article  CAS  Google Scholar 

  • Hyronimus B, Le Marrec C, Hadj Sassi A, and Deschamps A (2000) Acid and bile tolerance of spore-forming lactic acid bacteria. Int J Food Microbiol 61, 193–7.

    Article  CAS  Google Scholar 

  • Jang SJ, Kim YJ, Park JM, and Park YS (2011) Analysis of microflora in gochujang, Korean traditional fermented food. Food Science and Biotechnology 20, 1435–40.

    Article  Google Scholar 

  • Kakino M, Izuta H, Tsuruma K, Araki Y, Shimazawa M, Ichihara K, and Hara H (2012) Laxative effects and mechanism of action of Brazilian green propolis. BMC Complem Altern Med 12, 192.

    Article  Google Scholar 

  • Katzung BG (2004) In Basic and Clinical Pharmacology, (9th ed.), Lange Medical Books, USA.

    Google Scholar 

  • Kim TW, Lee JH, Kim SE, Park MH, Chang HC, and Kim HY (2009) Analysis of microbial communities in doenjang, a Korean fermented soybean paste, using nested PCR-denaturing gradient gel electrophoresis. Int J Food Microbiol 131, 265–71.

    Article  CAS  Google Scholar 

  • Klaenhammer TR and Kullen MJ (1999) Selection and design of probiotics. Int J Food Microbiol 50, 45–57.

    Article  CAS  Google Scholar 

  • Lee DK, Jang S, Baek EH, Kim MJ, Lee KS, Shin HS, Chung MJ, Kim JE, Lee KO, and Ha NJ (2009) Lactic acid bacteria affect serum cholesterol levels, harmful fecal enzyme activity, and fecal water content. Lipids Health Dis 8, 21.

    Article  Google Scholar 

  • Lee HY, Kim JH, Jeung HW, Lee CU, Kim DS, Li B, Lee GH, Sung MS, Ha KC, Back HI, Kim SH, Park SH, Oh MR, Kim MG, Jeon JY, Im YJ, Hwang MH, So BO, Shin SJ, Yoo WH, Kim HR, and Chae HJ (2012) Effects of Ficuscarica paste on loperamide-induced constipation in rats. Food Chem Toxicol 50, 895–902.

    Article  CAS  Google Scholar 

  • Lee JS, Heo GY, Lee JW, Oh YJ, Park JA, Park YH, Pyun YR, and Ahn JS (2005) Analysis of kimchi microflora using denaturing gradient gel electrophoresis. Int J Food Microbiol 102, 143–50.

    Article  CAS  Google Scholar 

  • Leung FW (2007) Etiologic factors chronic constipation: review of the scientific evidence. Dig Dis Sci 52, 313–6.

    Article  Google Scholar 

  • Lian WC, Hsiao HC, and Chou CC (2003) Viability of microencapsulated bifidobacteria in simulated gastric juice and bile solution. Int J Food Microbiol 86, 293–301.

    Article  Google Scholar 

  • Lilly DM and Stillwell RH (1965) Probiotics: growth-promoting factors produced by microorganisms. Science, 147, 747–8.

    Article  CAS  Google Scholar 

  • Lim SM and Im DS (2009) Screening and characterization of probiotic lactic acid bacteria isolated from Korean fermented foods. J Microbiol Biotechnol 19, 178–86.

    Article  CAS  Google Scholar 

  • Lin H, Prather C, Fisher R, Meyer J, Summers R, Pimentel M, Mccallum R, Akkermans L, and Loening-Baucke M (2005) Measurement of gastrointestinal transit. Dig Dis Sci 50, 989–1004.

    Article  Google Scholar 

  • Miki T, Minami K, Shinozaki H, Matsumura K, Saraya A, Ikeda H, Yamada Y, Holst JJ, and Seino S (2005) Distinct effects of glucose-dependent insulinotropic polypeptide and glucagon-like peptide-1 on insulin secretion and gut motility. Diabetes 54, 1056–63.

    Article  CAS  Google Scholar 

  • Mitsui N, Tsukahara M, Murasawa H, Tamura M, Kajimoto O, and Nishimura A (2006) Effect of natto including Bacillus subtilis K-2 (spore) on defecation and fecal microbiota, and safety of excessive ingestion in healthy volunteers. Yakuri to chiryo 34, 135–48.

    Google Scholar 

  • Nam YD, Lee SY, and Lim SI (2012a) Microbial community analysis of Korean soybean pastes by next-generation sequencing. Int J Food Microbiol 155, 36–42.

    Article  Google Scholar 

  • Nam YD, Yi SH, and Lim SI (2012b) Bacterial diversity of cheonggukjang, a traditional Korean fermented food, analyzed by barcoded pyrosequencing. Food Control 28, 135–42.

    Article  Google Scholar 

  • Ouwehand AC, Salminen S, and Isolauri E (2002) Probiotics: an overview of beneficial effects. Antonie Van Leeuwenhoek 82, 279–89.

    Article  CAS  Google Scholar 

  • Ouwehand AC, Tuomola EM, Tölkkö S, and Salminen S (2001) Assessment of adhesion properties of novel probiotic strains to human intestinal mucus. Int J Food Microbiol 64, 119–26.

    Article  CAS  Google Scholar 

  • Padmanabhan P, Grosse J, Asad AB, Radda GK, and Golay X (2013) Gastrointestinal transit measurements in mice with 99mTc-DTPA-labeled activated charcoal using NanoSPECT-CT. EJNMMI Research 3, 60

    Article  Google Scholar 

  • Peypoux F, Guinand M, Michel G, Delcambe L, Das BC, and Lederer E (1978) Structure of iturine A, a peptidolipid antibiotic from Bacillus subtilis. Biochemistry 17, 3992–6.

    Article  CAS  Google Scholar 

  • Rowan NJ, Deans K, Anderson JG, Gemmell CG, Hunter IS, and Chaithong T (2001) Putative virulence factor expression by clinical and food isolates of Bacillus spp. after growth in reconstituted infant milk formulae. Appl Environ Microbiol 67, 3873–81.

    Article  CAS  Google Scholar 

  • Rubio R, Aymerich T, Bover-Cid S, Guàrdia MD, Arnau J, and Garriga M (2013) Probiotic strains Lactobacillus plantarum 299V and Lactobacillus rhamnosus GG as starter cultures for fermented sausages. LWT-Food Sci Technol 54, 51–6

    Article  CAS  Google Scholar 

  • Sanders ME, Morelli L, and Tompkins TA (2003) Sporeformers as human probiotics: Bacillus, Sporolactobacillus, and Brevibacillus. Compr Rev Food Sci F 2, 101–110.

    Article  Google Scholar 

  • Schiller L (2001) The therapy of constipation. Aliment Pharm Therap 15, 749–63.

    Article  CAS  Google Scholar 

  • Schiller LR, Santa Ana CA, Morawski SG, and Fordtran JS (1984) Mechanism of the antidiarrheal effect of loperamide. Gastroenterology 86, 1475–83.

    CAS  Google Scholar 

  • Schrezenmeir J and de Vrese M (2001) Probiotics, prebiotics, and synbioticsapproaching a definition. AM J Clin Nutr 73, 361S–4S.

    CAS  Google Scholar 

  • Servin AL and Coconnier MH (2003) Adhesion of probiotic strains to the intestinal mucosa and interaction with pathogens. Best Pract Res Clin Gastroenterol 17, 741–54.

    Article  CAS  Google Scholar 

  • Shimotoyodome A, Meguro S, Hase T, Tokimitsu I, and Sakata T (2000) Decreased colonic mucus in rat with loperamide-induced constipation. Comp Biochem Physiol A Mol Integr Physiol 126, 203–11.

    Article  CAS  Google Scholar 

  • Sorokulova IB, Pinchuk IV, Denayrolles M, Osipova IG, Huang JM, Cutting SM, and Urdaci MC (2008) The safety of two Bacillus probiotic strains for human use. Dig Dis Sci 53, 954–63.

    Article  Google Scholar 

  • Stanton C, Gardiner G, Meehan H, Collins K, Fitzgerald G, Lynch PB, and Ross RP (2001) Market potential for probiotics. AM J Clin Nutr 73, 476S–83S.

    CAS  Google Scholar 

  • Stein T (2005) Bacillus subtilis antibiotics: structures, syntheses and specific functions. Mol Microbiol 56, 845–57.

    Article  CAS  Google Scholar 

  • Stenfors Arnesen LP, Fagerlund A, and Granum PE (2008) From soil to gut: Bacillus cereus and its food poisoning toxins. FEMS Microbiol Rev 32, 579–606.

    Article  CAS  Google Scholar 

  • Tan-No K, Niijima F, Nakagawasai O, Sato T, Satoh S, and Tadano T (2003) Development of tolerance to the inhibitory effect of loperamide on gastrointestinal transit in mice. Eur J Pharm Sci 20, 357–63.

    Article  CAS  Google Scholar 

  • Vinderola CG and Reinheimer JA (2003) Lactic acid starter and probiotic bacteria: a comparative “in vitro” study of probiotic characteristics and biological barrier resistance. Food Rev Int 36, 895–904.

    Article  CAS  Google Scholar 

  • Walia R, Mahajan L, and Steffen R (2009) Recent advances in chronic constipation. Curr Opin Pediatr 21, 661–6.

    Article  Google Scholar 

  • Wang B, Wei H, Yuan J, Li Q, Li Y, Li N, and Li J (2008) Identification of a surface protein from Lactobacillus reuteri JCM1081 that adheres to porcine gastric mucin and human enterocyte-like HT-29 cells. Curr microbiol 57, 33–8.

    Article  CAS  Google Scholar 

  • Wang ZT, Xiao GX, Yao YM, Guo SZ, Lu KH, and Sheng ZY (2006) The role of bifidobacteria in gut barrier function after thermal injury in rats. J Trauma 61, 650–7.

    Article  Google Scholar 

  • Wang ZT, Yao YM, Xiao GX, and Sheng ZY (2004) Risk factors of development of gut-derived bacterial translocation in thermally injured rats. World J Gastroenterol 10, 1619–24.

    Google Scholar 

  • Wintola OA, Sunmonu TO, and Afolayan AJ (2010) The effect of Aloe ferox Mill. in the treatment of loperamide-induced constipation in Wistar rats. BMC Gastroenterol 10, 95.

    Article  Google Scholar 

  • Ziaei-Nejad S, Rezaei M H, Takami GA, Lovett DL, Mirvaghefi AR, and Shakouri M (2006) The effect of Bacillus spp. bacteria used as probiotics on digestive enzyme activity, survival and growth in the Indian white shrimp Fenneropenaeus indicus. Aquaculture 252, 516–24.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biohealth Convergence Center, Daegu Technopark, Seong-seo Industrial complex-ro, Dalseo-gu, Daegu, 704-801, Republic of Korea

    Bae Jin Kim, Yoo Seok Jeong & Hee Kyoung Jung

  2. Department of Food Science and Technology, Catholic University of Daegu, Geumnakro 5, Hayang-Eup, Gyeongsan-Si, Gyeongbuk, Republic of Korea

    Joo-Heon Hong

Authors
  1. Bae Jin Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Joo-Heon Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yoo Seok Jeong
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hee Kyoung Jung
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hee Kyoung Jung.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kim, B.J., Hong, JH., Jeong, Y.S. et al. Evaluation of two Bacillus subtilis strains isolated from Korean fermented food as probiotics against loperamide-induced constipation in mice. J Korean Soc Appl Biol Chem 57, 797–806 (2014). https://doi.org/10.1007/s13765-014-4106-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13765-014-4106-0

Keywords

Search

Navigation