Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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In Vitro Characterization of Lactic Acid Bacteria from Indonesian Kefir Grains as Probiotics with Cholesterol-Lowering Effect

  • Yusuf, Dandy (Department of Food Science and Technology, Faculty of Agricultural Technology, IPB University (Bogor Agricultural University)) ;
  • Nuraida, Lilis (Department of Food Science and Technology, Faculty of Agricultural Technology, IPB University (Bogor Agricultural University)) ;
  • Dewanti-Hariyadi, Ratih (Department of Food Science and Technology, Faculty of Agricultural Technology, IPB University (Bogor Agricultural University)) ;
  • Hunaefi, Dase (Department of Food Science and Technology, Faculty of Agricultural Technology, IPB University (Bogor Agricultural University))
  • Received : 2019.10.14
  • Accepted : 2020.01.21
  • Published : 2020.05.28
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Abstract

Indonesian kefir grains are potential sources of lactic acid bacteria (LAB) that may act as probiotics with specific functional properties. In this study we explored the potential of the probiotic and cholesterol-lowering effect of LAB isolated from Indonesian kefir grains obtained from Bogor, Bandung, Jakarta, and Yogyakarta. The results revealed that 10 isolates showed considerable survivability at low pH and bile salt with total cell reduction of ~3 log colony-forming units per milliliter after exposure to pH 2.5 and 0.5% (w/v) bile salt for 1 and 3 h, respectively. All strains exhibited strong antimicrobial activities against pathogenic bacteria and were sensitive to a wide spectrum of antibiotics but exhibited weak bile salt hydrolase activity. Identification based on 16S RNA suggested that nine isolates were Lactobacillus kefiri and one was Lactobacillus rhamnosus. The ability of the isolates to reduce cholesterol from the media varied, ranging from 22.08% to 68.75% with the highest reduction shown by L. kefiri JK17. The ability to remove cholesterol from the media decreased greatly in resting and dead cells, ranging from 14.58% to 22.08% in resting cells and from 7.89% to 18.17% in dead cells. It can be concluded that Indonesian kefir grains contain LAB potentially acting as probiotics capable of reducing cholesterol. The cholesterol-lowering effect especially occurs when the cells are metabolically active.

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References

  1. Hertzler SR, Clancy SM. 2003. Kefir improves lactose digestion and tolerance in adults with lactose maldigestion. J. Am. Diet. Assoc. 103: 582-587. https://doi.org/10.1053/jada.2003.50111
  2. Kim DH, Jeong D, Kim H, Kang IB, Chon JW, Song KY, et al. 2016. Antimicrobial activity of kefir against various food pathogens and spoilage bacteria. Korean J. Food Sci. Anim. Resour. 36: 787-790. https://doi.org/10.5851/kosfa.2016.36.6.787
  3. Ozcan A, Kaya N, Atakisi O, Karapehlivan M, Atakisi E, Cenesiz S. 2009. Effect of kefir on the oxidative stress due to lead in rats. J. Appl. Anim. Res. 35: 91-93. https://doi.org/10.1080/09712119.2009.9706992
  4. Liu JR, Wang SY, Chen MJ, Chen HL, Yueh PY, Lin CW. 2006. Hypocholesterolaemic effects of milk kefir and soyamilk kefir in cholesterol-fed hamsters. Br. J. Nutr. 95: 939-946. https://doi.org/10.1079/BJN20061752
  5. Hadisaputro S, Djokomoeljanto RRJ, Judiono, Soesatyo MHNE. 2012. The effects of oral plain kefir supplementation on proinflammatory cytokine properties of the hyperglycemia Wistar rats induced by streptozotocin. Acta Med. Indones. 44: 100-104.
  6. Halloran K, Underwood MA. 2019. Probiotic mechanisms of action. Early Hum. Dev. 135: 58-65. https://doi.org/10.1016/j.earlhumdev.2019.05.010
  7. Chaves-Lopez C, Serio A, Paparella A, Martuscelli M, Corsetti A, Tofalo R, et al. 2014. Impact of microbial cultures on proteolysis and release of bioactive peptides in fermented milk. Food Microbiol. 42: 117-121. https://doi.org/10.1016/j.fm.2014.03.005
  8. FAO/WHO. 2002. Working group report on drafting guidelines for the evaluation of probiotics in food. World Health Organization and Food and Agriculture Organization of the United Nations. Ontario: London.
  9. Bhatnagar D, Soran H, Durrington PN. 2008. Hypercholesterolaemia and its management. BMC 337: a993. https://doi.org/10.1136/bmj.a993
  10. Choi EA, Chang HC. 2015. Cholesterol-lowering effects of a putative probiotic strain Lactobacillus plantarum EM isolated from kimchi. LWT - Food Sci. Technol. 62: 210-217. https://doi.org/10.1016/j.lwt.2015.01.019
  11. Tomaro-Duchesneau C, Jones ML, Shah D, Jain P, Saha S, Prakash S. 2014. Cholesterol assimilation by lactobacillus probiotic bacteria: an in vitro investigation. Biomed. Res. Int. 2014.
  12. Kim DH, Jeong D, Kang IB, Kim H, Song KY, Seo KH. 2017. Dual function of Lactobacillus kefiri DH5 in preventing high-fat-dietinduced obesity: direct reduction of cholesterol and upregulation of PPAR-$\alpha$ in adipose tissue. Mol. Nutr. Food Res. 61: 1-35.
  13. Miremadi F, Ayyash M, Sherkat F, Stojanovska L. 2014. Cholesterol reduction mechanisms and fatty acid composition of cellular membranes of probiotic Lactobacilli and Bifidobacteria. J. Funct. Foods 9: 295-305. https://doi.org/10.1016/j.jff.2014.05.002
  14. Anila K, Kunzez A, Bhalla TC. 2016. In vitro cholesterol assimilation and functional enzymatic activities of putative probiotic Lactobacillus sp. isolated from fermented foods/ beverages of north west India. J. Nutr. Food Sci. 6: 2.
  15. Lye HS, Rusul G, Liong MT. 2010. Removal of cholesterol by Lactobacilli via incorporation and conversion to coprostanol. J. Dairy Sci. 93: 1383-1392. https://doi.org/10.3168/jds.2009-2574
  16. Gilliland SE, Nelson CR, Maxwell C. 1985. Assimilation of cholesterol by Lactobacillus acidophilus. Appl. Environ. Microbiol. 49: 377-381. https://doi.org/10.1128/aem.49.2.377-381.1985
  17. Guo CF, Zhang LW, Han X, Li JY, Du M, Yi HX, et al. 2011. Short communication: A sensitive method for qualitative screening of bile salt hydrolase-active lactobacilli based on thin-layer chromatography. J. Dairy Sci. 94: 1732-1737. https://doi.org/10.3168/jds.2010-3801
  18. Wang SC, Chang CK, Chan SC, Shieh JS, Chiu CK, Duh PD. 2014. Effects of lactic acid bacteria isolated from fermented mustard on lowering cholesterol. Asian Pac. J. Trop. Biomed. 4: 523-528. https://doi.org/10.12980/APJTB.4.201414B54
  19. Shehata MG, El Sohaimy SA, El-Sahn MA, Youssef MM. 2016. Screening of isolated potential probiotic lactic acid bacteria for cholesterol lowering property and bile salt hydrolase activity. Ann. Agric. Sci. 61: 65-75. https://doi.org/10.1016/j.aoas.2016.03.001
  20. Nuraida L. 2015. A review?: Health promoting lactic acid bacteria in traditional Indonesian fermented foods. Food Sci. Hum. Wellness. 4: 47-55. https://doi.org/10.1016/j.fshw.2015.06.001
  21. Teneva-angelova T, Hristova I, Pavlov A. 2018. Lactic acid bacteria - from nature through food to health, pp. 91-133. In the Handbook of Food Bioengineering, Advances in Biotechnology for Food Industry, 14th Ed. Academic Press. Elsevier.
  22. Liasi SA, Azmi TI, Hassan MD, Shuhaimi M, Rosfarizan M, Ariff AB. 2009. Antimicrobial activity and antibiotic sensitivity of three isolates of lactic acid bacteria from fermented fish product, Budu. Malays. J. Microbiol. 5: 33-37.
  23. Ryu EH, Chang HC. 2013. In vitro study of potentially probiotic lactic acid bacteria strains isolated from kimchi. Ann. Microbiol. 63: 1387-1395. https://doi.org/10.1007/s13213-013-0599-8
  24. EFSA. 2010. Guidance on the risk assessment of genetically modified microorganisms and their food and feed products. EFSA J. 1-67.
  25. Zanirati DF, Abatemarco M, Sandes SH de C, Nicoli JR, Nunes AC, Neumann E. 2015. Selection of lactic acid bacteria from Brazilian kefir grains for potential use as starter or probiotic cultures. Anaerobe 32: 70-76. https://doi.org/10.1016/j.anaerobe.2014.12.007
  26. Rudel LL, Morris MD. 1973. Determination of cholesterol using o-phthalaldehyde. J. Lipid Res. 14: 364-6. https://doi.org/10.1016/S0022-2275(20)36896-6
  27. Cotter PD, Hill C. 2003. Surviving the acid test: Responses of Gram-Positive bacteria to low pH. Microbiol. Mol. Biol. Rev. 67: 429-453. https://doi.org/10.1128/MMBR.67.3.429-453.2003
  28. Tokatl M, Gulgor G, Elmac SB, Isleyen NA, Ozcelik F. 2015. In Vitro properties of potential probiotic indigenous lactic acid bacteria originating from traditional pickles. Biomed. Res. Int. 2015: 1-8.
  29. Jacobsen CN, Nielsen VR, Hayford AE, Moller PL, Michaelsen KF, Pærregaard A, Sandstrom B, Tvede M, Jakobsen M. 1999. Screening of probiotic activities of forty-seven strains of Lactobacillus spp. by in vitro techniques and evaluation of the colonization ability of five selected strains in humans. Appl. Environ. Microbiol. 65: 4949-4956. https://doi.org/10.1128/aem.65.11.4949-4956.1999
  30. Kimoto H, Kurisaki J, Tsuji NM, Ohmomo S, Okamoto T. 1999. Lactococci as probiotic strains: Adhesion to human enterocyte-like Caco-2 cells and tolerance to low pH and bile. Lett. Appl. Microbiol. 29: 313-316. https://doi.org/10.1046/j.1365-2672.1999.00627.x
  31. Begley M, Gahan CGM, Hill C. 2005. The interaction between bacteria and bile. FEMS Microbiol. Rev. 29: 625-651. https://doi.org/10.1016/j.femsre.2004.09.003
  32. De Smet I, Van Hoorde L, Vande Woestyne M, Christiaens H, Verstraete W. 1995. Significance of bile salt hydrolytic activities of lactobacilli. J. Appl. Bacteriol. 79: 292-301. https://doi.org/10.1111/j.1365-2672.1995.tb03140.x
  33. Sarkar S, Sur A, Sarkar K, Majhi R, Basu S, Chatterjee K, Sikder B. 2016. Probiotics: A way of value addition in functional food. Int. J. Food Sci. Nutr. Diet. 5: 290-293.
  34. Broaders E, Gahan CGM, Marchesi JR. 2013. Potential for spread of antibiotic resistance genes mobile genetic elements of the human gastrointestinal tract. Gut Microbes. 4: 271-280. https://doi.org/10.4161/gmic.24627
  35. Guo L, Wang L, Liu F, Li B, Tang Y, Yu S, Zhang D. 2019. Effect of bile salt hydrolase-active Lactobacillus plantarum KLDS 1.0344 on cholesterol metabolism in rats fed a high-cholesterol diet. J. Funct. Foods 61:103497. https://doi.org/10.1016/j.jff.2019.103497
  36. Clarridge JE. 2004. Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clin. Microbiol. Rev. 17: 840-862. https://doi.org/10.1128/CMR.17.4.840-862.2004
  37. Horiike T, Miyata D, Hamada K, Saruhashi S, Shinozawa T, Kumar S, Chakraborty R, Komiyama T, Tateno Y. 2009. Phylogenetic construction of 17 bacterial phyla by new method and carefully selected orthologs. Gene 429: 59-64. https://doi.org/10.1016/j.gene.2008.10.006
  38. Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol. Biol. Evol. 24: 1596-1599. https://doi.org/10.1093/molbev/msm092
  39. Maryati Y, Nuraida L, Dewanti-Hariyadi R. 2016. A study in vitro of latic acid bacteria (LAB) isolates on cholesterol lowering ability in the presence of oligosaccharides. Agritech. 36: 196-205. https://doi.org/10.22146/agritech.12865
  40. Kimoto H, Ohmomo S, Okamoto T. 2002. Cholesterol removal from media by Lactococci. J. Dairy Sci. 85: 3182-3188. https://doi.org/10.3168/jds.S0022-0302(02)74406-8

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