Isolation and Identification of Lactic Acid Bacteria from Okara and Evaluation of Their Potential as Candidate Probiotics
DOI:
https://doi.org/10.3923/pjn.2017.618.628Keywords:
Antibacterial activity, bile salt, lactic acid bacteria, Okara, pathogenic bacteriaAbstract
Background: Okara is an industrial waste product resulting from the manufacture of soy milk or tofu. This waste is a pulp consisting of the insoluble parts of the soybean that remain after pureed soybeans are filtered during the production of soy milk and tofu. Okara is not widely used and is dumped into the environment, causing pollution. However, this waste contains nutrients and may be a good medium for the growth and development of microorganisms such as Lactic Acid Bacteria (LAB). Objective: The purpose of this study was to isolate and identify probiotic LAB from okara and determine their antimicrobial activity against pathogenic bacteria (Escherichia coli 0157, Staphylococcus aureus ATCC 25923 and Listeria monocytogenes EP01). Methodology: Isolation and purification of LAB were carried out on MRS agar supplemented with 0.2% CaCO3. Isolates were identified by morphological and physiological tests. Antibacterial activity was tested using the well diffusion method and the obtained data were analyzed descriptively. Twenty-four isolates were grown on MRS agar. Results: Sixteen isolates that exhibited growth at the clear zone were identified as gram positive, catalase-negative homofermentative rod and coccus cells. The results showed that the LAB isolated from okara had the ability to inhibit pathogenic bacteria, causing various clear zone diameters. The survival rates of these isolates under acidic conditions and their tolerance to 0.5% bile salt were varied. Based on the antimicrobial activity and probiotic properties of the LAB isolated from okara, these isolates belong to the species Lactobacillus spentosus and Lactobacillus plantarum. Conclusion: The results showed that the LAB isolated from okara belonged to the species Lactobacillus pentosus and Lactobacillus plantarum. Both species exhibited properties attributed to probiotics.
References
Suruga, K., A. Kato, K. Kadokura, W. Hiruma, Y. Sekino, C.A.T. Buffington and K. Komatsu, 2011. "Okara" A New Preparation of Food Material with Antioxidant Activity and Dietary Fiber from Soybean. In: Soybean and Nutrition, El-Shemy, H. (Ed.). InTech Publisher, Croatia, ISBN: 978-953-307-536-5, pp: 311-326.
Kitawaki, R., Y. Nishimura, N. Takagi, M. Iwasaki, K. Tsuzuki and M. Fukuda, 2009. Effects of lactobacillus fermented soymilk and soy yogurt on hepatic lipid accumulation in rats fed a cholesterol-free diet. Biosci. Biotechnol. Biochem., 73: 1484-1488.
Hsieh, C. and F.C. Yang, 2003. Reusing soy residue for the solid-state fermentation of Ganoderma lucidum. Bioresour. Technol., 91: 105-109.
Marazza, J.A., J.G. LeBlanc, G.S. de Giori and M.S. Garro, 2013. Soymilk fermented with Lactobacillus rhamnosus CRL981 ameliorates hyperglycemia, lipid profiles and increases antioxidant enzyme activities in diabetic mice. J. Funct. Foods, 5: 1848-1853.
Oz, E., G. Kaban, O. Baris and M. Kaya, 2017. Isolation and identification of lactic acid bacteria from pastırma. Food Control, 77: 158-162.
Visintin, S., V. Alessandria, A. Valente, P. Dolci and L. Cocolin, 2016. Molecular identification and physiological characterization of yeasts, lactic acid bacteria and acetic acid bacteria isolated from heap and box cocoa bean fermentations in West Africa. Int. J. Food Microbiol., 216: 69-78.
Reis, N.A., M.A.F. Saraiva, E.A.A. Duarte, E.A. Carvalho, B.B. Vieira and N.S. Evangelistaâ€Barreto, 2016. Probiotic properties of lactic acid bacteria isolated from human milk. J. Applied Microbiol., 121: 811-820.
Leite, A.M.O., M.A.L. Miguel, R.S. Peixoto, P. Ruas-Madiedo, V.M.F. Paschoalin, B. Mayo and S. Delgado, 2015. Probiotic potential of selected lactic acid bacteria strains isolated from Brazilian kefir grains. J. Dairy Sci., 98: 3622-3632.
Ni, K., Y. Wang, D. Li, Y. Cai and H. Pang, 2015. Characterization, identification and application of lactic acid bacteria isolated from forage paddy rice silage. PLoS ONE, Vol. 10.
Ogunremi, O.R., A.I. Sanni and R. Agrawal, 2015. Probiotic potentials of yeasts isolated from some cerealâ€based Nigerian traditional fermented food products. J. Applied Microbiol., 119: 797-808.
Ouwehand, A.C., S. Salminen and E. Isolauri, 2002. Probiotics: An Overview of Beneficial Effects. In: Lactic Acid Bacteria: Genetics, Metabolism and Applications, Siezen, R.J., J. Kok, T. Abee and G. Schaafsma (Eds.). Springer, Netherlands, ISBN: 978-1-4020-0922-8, pp: 279-289.
Fontana, L., M. Bermudez-Brito, J. Plaza-Diaz, S. Munoz-Quezada and A. Gil, 2013. Sources, isolation, characterisation and evaluation of probiotics. Br. J. Nutr., 109: S35-S50.
Gourbeyre, P., S. Denery and M. Bodinier, 2011. Probiotics, prebiotics and synbiotics: Impact on the gut immune system and allergic reactions. J. Leukocyte Biol., 89: 685-695.
Frick, J.S., K. Schenk, M. Quitadamo, F. Kahl and M. Koberle et al., 2007. Lactobacillus fermentum attenuates the proinflammatory effect of Yersinia enterocolitica on human epithelial cells. Inflamm. Bowel Dis., 13: 83-90.
Akbar, A., I. Ali and A.K. Anal, 2016. Industrial perspectives of lactic acid bacteria for biopreservation and food safety. J. Anim. Plant Sci., 26: 938-948.
Cleveland, J., T.J. Montville, I.F. Nes and M.L. Chikindas, 2001. Bacteriocins: Safe, natural antimicrobials for food preservation. Int. J. Food Microbiol., 71: 1-20.
Mazzoli, R., F. Bosco, I. Mizrahi, E.A. Bayer and E. Pessione, 2014. Towards lactic acid bacteria-based biorefineries. Biotechnol. Adv., 32: 1216-1236.
Awojobi, K.O., S.M. Adeyemo and O.O. Sanusi, 2016. Biosynthesis of antimicrobial compounds by lactic acid bacteria and its use as biopreservative in pineapple juice. Front. Sci., 6: 17-24.
Marshall, S.H. and G. Arenas, 2003. Antimicrobial peptides: A natural alternative to chemical antibiotics and a potential for applied biotechnology. Electron. J. Biotechnol., 6: 271-284.
Ammor, S., G. Tauveron, E. Dufour and I. Chevallier, 2006. Antibacterial activity of lactic acid bacteria against spoilage and pathogenic bacteria isolated from the same meat small-scale facility: 1-Screening and characterization of the antibacterial compounds. Food Control, 17: 454-461.
Arques, J.L., E. Rodriguez and S. Langa, 2015. Antimicrobial activity of lactic acid bacteria in dairy products and gut: Effect on pathogens. BioMed Res., Vol. 2015.
Rosa, P.R.F., B.C. Gomes, M.B.A. Varesche and E.L. Silva, 2016. Characterization and antimicrobial activity of lactic acid bacteria from fermentative bioreactors during hydrogen production using cassava processing wastewater. Chem. Eng. J., 284: 1-9.
Siroli, L., F. Patrignani, D.I. Serrazanetti, G. Tabanelli, C. Montanari, F. Gardini and R. Lanciotti, 2015. Lactic acid bacteria and natural antimicrobials to improve the safety and shelf-life of minimally processed sliced apples and lamb's lettuce. Food Microbiol., 47: 74-84.
Li, Q., 2015. Antimicrobial activity of lactic acid bacteria isolated from fermented Taro Skins against Salmonella typhimurium and Listeria monocytogenes. Proceedings of the International Association for Food Protection Annual Meeting, July 25-28, 2015, Portland, Oregon, USA.
Joo, N.E., K. Ritchie, P. Kamarajan, D. Miao and Y.L. Kapila, 2012. Nisin, an apoptogenic bacteriocin and food preservative, attenuates HNSCC tumorigenesis via CHAC1. Cancer Med., 1: 295-305.
Chen, Y.S., F. Yangida and T. Shinahara, 2005. Isolation and identification of lactic acid bacteria from soil using an enrichment procedure. Lett. Applied Microbiol., 40: 195-200.
Felten, A., C. Barreau, C. Bizet, P.H. Lagrange and A. Philippon, 1999. Lactobacillus species identification, H2O2 production and antibiotic resistance and correlation with human clinical status. J. Clin., 37: 729-733.
Holzapfel, W.H. and B.J. Wood, 2014. Lactic Acid Bacteria: Biodiversity and Taxonomy. John Wiley and Sons, New York, ISBN: 9781444333831, Pages: 632.
Roberts, D., W. Hooper, M. Greenwood and C. Struijk, 1996. Practical Food Microbiology. 3rd Edn., Blackwell Publishing, Oxford.
Salminen, S., A. Ouwehand, Y. Benno and Y.K. Lee, 1999. Probiotics: How should they be defined? Trends Food Sci. Technol., 10: 107-110.
Bromberg, R., I. Moreno, C.L. Zaganini, R.R. Delboni and J. de Oliveira, 2004. Isolation of bacteriocin-producing lactic acid bacteria from meat and meat products and its spectrum of inhibitory activity. Braz. J. Microbiol., 35: 137-144.
Hawaz, E., 2014. Isolation and identification of probiotic lactic acid bacteria from curd and in vitro evaluation of its growth inhibition activities against pathogenic bacteria. Afr. J. Microbiol. Res., 8: 1419-1425.
Oluwajoba, S.O., F.A. Akinyosoye and V.O. Oyetayo, 2013. In vitro screening and selection of probiotic lactic acid bacteria isolated from spontaneously fermenting Kunu-zaki. Adv. Microbiol., 3: 309-316.
Seeley, H.W., P.J. VanDemark and J.J. Lee, 1990. Selected Exercises for Microbes in Action. 4th Edn., W.H. Freeman & Co. Ltd., New York, ISBN: 9780716721116, Pages: 150.
Lu, F., Y. Liu and B. Li, 2013. Okara dietary fiber and hypoglycemic effect of Okara foods. Bioactive Carbohydr. Dietary Fibre, 2: 126-132.
Holzapfel, W.H.N. and B.J.B. Wood, 2012. The Genera of Lactic Acid Bacteria. Springer, Netherlands, ISBN: 9781461558170, Pages: 398.
Roberts, D. and M. Greenwood, 2003. Practical Food Microbiology. 3rd Edn., Blackwell Publishing Ltd., USA., ISBN-13: 9781405100755, Pages: 294.
Salminen, S. and A.V. Wright, 2004. Lactic Acid Bacteria: Microbiological and Functional Aspects. 2nd Edn., Marcell Dekker, New York.
Garbutt, J., 1997. Essentials of Food Microbiology. Arnold Publishers, London.
Allen, S.J., E.G. Martinez, G.V. Gregorio and L.F. Dans, 2010. Probiotics for treating acute infectious diarrhoea. Cochrane Database Syst. Rev.
Savadogo, A., C.A.T. Ouattara, I.H.N. Bassole and A.S. Traore, 2004. Antimicrobial activities of lactic acid bacteria strains isolated from Burkina Faso fermented milk. Pak. J. Nutr., 3: 174-179.
Arief, I.I., B.S.L. Jenie, M. Astawan, K. Fujiyama and A.B. Witarto, 2015. Identification and probiotic characteristics of lactic acid bacteria isolated from Indonesian local beef. Asian J. Anim. Sci., 9: 25-36.
Jawetz, E., J. Melnick, E. Adelberg and G. Brooks, 1996. Mikrobiologi Kedokteran. EGC, Penerbit Buku Kedokteran, Jakarta, Indonesia.
Alakomi, H.L., E. Skytta, M. Saarela, T. Mattila-Sandholm, K. Latva-Kala and I.M. Helander, 2000. Lactic acid permeabilizes gram-negative bacteria by disrupting the outer membrane. Applied Environ. Microbiol., 66: 2001-2005.
Oyetayo, V.O., F.C. Adetuyi and F.A. Akinyosoye, 2003. Safety and protective effect of Lactobacillus acidophilus and Lactobacillus casei used as probiotic agent in vivo. Afr. J. Biotechnol., 2: 448-452.
Schillinger, U. and F. Lucke, 1989. Antibacterial activity of Lactobacillus sake isolated from meat. Applied Environ. Microbiol., 55: 1901-1906.
Messaoudi, S., M. Manai, G. Kergourlay, H. Prevost, N. Connil, J.M. Chobert and X. Dousset, 2013. Lactobacillus salivarius: Bacteriocin and probiotic activity. Food Microbiol., 36: 296-304.
Mansour, N.M., H. Heine, S.M. Abdou, M.E. Shenana, M.K. Zakaria and A. Elâ€Diwany, 2014. Isolation of Enterococcus faecium NM113, Enterococcus faecium NM213 and Lactobacillus casei NM512 as novel probiotics with immunomodulatory properties. Microbiol. Immunol., 58: 559-569.
Novak, J. and J.A. Katz, 2006. Probiotics and prebiotics for gastrointestinal infections. Curr. Infect. Dis. Rep., 8: 103-109.
Delgado, A., D. Brito, C. Peres, F. Noe-Arroyo and A. Garrido-Fernandez, 2005. Bacteriocin production by Lactobacillus pentosus B96 can be expressed as a function of temperature and NaCl concentration. Food Microbiol., 22: 521-528.
Siragusa, S., M. de Angelis, M. Calasso, D. Campanella, F. Minervini, R. di Cagno and M. Gobbetti, 2014. Fermentation and proteome profiles of Lactobacillus plantarum strains during growth under food-like conditions. J. Proteomics, 96: 366-380.
Blana, V.A., A. Grounta, C.C. Tassou, G.J.E. Nychas and E.Z. Panagou, 2014. Inoculated fermentation of green olives with potential probiotic Lactobacillus pentosus and Lactobacillus plantarum starter cultures isolated from industrially fermented olives. Food Microbiol., 38: 208-218.
Ogunbanwo, S.T., A.I. Sanni and A.A. Onilude, 2003. Characterization of bacteriocin produced by Lactobacillus plantarum F1 and Lactobacillus brevis OG1. Afr. J. Biotechnol., 2: 219-227.
Montoro, B.P., N. Benomar, L.L. Lerma, S.C. Gutierrez, A. Galvez and H. Abriouel, 2016. Fermented alorena table olives as a source of potential probiotic Lactobacillus pentosus strains. Front. Microbiol., Vol. 7.
Khalid, K., 2011. An overview of lactic acid bacteria. Int. J. Biosci., 1: 1-13.
Argyri, A.A., G. Zoumpopoulou, K.A.G. Karatzas, E. Tsakalidou, G.J.E. Nychas, E.Z. Panagou and C.C. Tassou, 2013. Selection of potential probiotic lactic acid bacteria from fermented olives by in vitro tests. Food Microbiol. 33: 282-291.
Published
Issue
Section
License
Copyright (c) 2017 The Author(s)
This work is licensed under a Creative Commons Attribution 4.0 International License.
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited.
