Screening of Antagonistic/Synergistic Effect between Lactic Acid Bacteria (LAB) and Yeast Strains Isolated from Kefir
Kefir is a traditional fermented refreshing beverage which is known for its valuable and beneficial properties for human health. Mainly yeast species, lactic acid bacteria (LAB) strains and fewer acetic acid bacteria strains live together in a natural matrix named “kefir grain”, which is formed from various proteins and polysaccharides. Different microbial species live together in slimy kefir grain and it has been thought that synergetic effect could take place between microorganisms, which belong to different genera and species. In this research, yeast and LAB were isolated from kefir samples obtained from Uludag University Food Engineering Department. The cell morphology of isolates was screened by microscopic examination. Gram reactions of bacteria isolates were determined by Gram staining method, and as well catalase activity was examined. After observing the microscopic/morphological and physical, enzymatic properties of all isolates, they were divided into the groups as LAB and/or yeast according to their physicochemical responses to the applied examinations. As part of this research, the antagonistic/synergistic efficacy of the identified five LAB and five yeast strains to each other were determined individually by disk diffusion method. The antagonistic or synergistic effect is one of the most important properties in a co-culture system that different microorganisms are living together. The synergistic effect should be promoted, whereas the antagonistic effect is prevented to provide effective culture for fermentation of kefir. The aim of this study was to determine microbial interactions between identified yeast and LAB strains, and whether their effect is antagonistic or synergistic. Thus, if there is a strain which inhibits or retards the growth of other strains found in Kefir microflora, this circumstance shows the presence of antagonistic effect in the medium. Such negative influence should be prevented, whereas the microorganisms which have synergistic effect on each other should be promoted by combining them in kefir grain. Standardisation is the most desired property for industrial production. Each microorganism found in the microbial flora of a kefir grain should be identified individually. The members of the microbial community found in the glue-like kefir grain may be redesigned as a starter culture regarding efficacy of each microorganism to another in kefir processing. The main aim of this research was to shed light on more effective production of kefir grain and to contribute a standardisation of kefir processing in the food industry.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1129790Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1147
 B. Nielsen, G. C. Gürakan, “Kefir: A Multifaceted Fermented Dairy Product”, Probiotics and Antimicrobial Proteins, vol. 6, pp. 123-135, 2014.
 N. B. Gradova, A. A. Khokhlacheva, E. D. Murzina, V. V. Myasoyedova, “Microbial Components of Kefir Grains as Exopolysaccharide Kefiran Producers”, Applied Biochemistry and Microbiology, vol. 51(9), pp. 873–880, 2015.
 M.R. Prado, L. M. Blandon, L. P. S. Vandenberghe, C. Rodrigues, G. R. Castro, V. Thomaz-Soccol, C. R. Soccol, “Milk kefir: composition, microbial cultures, biological activities, and related products”, Frontiers in Microbiology, vol. 6, pp. 1-10, 2015.
 K. J. Heller, “Probiotic bacteria in fermented foods: product characteristics and starter organisms”, The American Journal of Clinical Nutrition, vol. 73(suppl), pp. 374-379, 2001.
 N. Habibi, S. Soleimanian-Zad, M. S. Zeinoddin, “Exopolysaccharides Produced by Pure Culture of Lactobacillus, Lactococcus and Yeast Isolated from Kefir Grain by Microtiter Plate Assay: Optimization and Comparison”, World Applied Sciences Journal, vol. 12 (6), pp. 742-750, 2011.
 L. Wang, H. Zhong, K. Liu, A. Guo, Z. Qi, M. Cai, “The evaluation of kefir pure culture starter: Liquid-core capsule entrapping microorganisms isolated from kefir grains”, Food Science and Technology International, vol. 22(7), pp. 598-608, 2015.
 O. Gul, M. Mortas, I. Atalar, M. Dervisoglu, T. Kahyaoglu, “Manufacture and characterization of kefir made from cow and buffalo milk, using kefir grain and starter culture”, J. Dairy Sci., vol. 98, pp. 1517–1525, 2015.
 B. Cheirsilp, S. Radchabut, “Use of whey lactose from dairy industry for economical kefiran production by Lactobacillus kefiranofaciens in mixed cultures with yeasts”, New Biotechnology, vol. 28(6), pp. 574-580, 2011.
 T. Pogačić, S. Šinko, Š. Zamberlin, D. Samaržija, “Microbiota of kefir grains”, Mljekarstvo, vol. 63(1), pp. 3-14, 2013.
 F. Mendes, S. Sieuwerts, E. de Hulster, M. J. H. Almering, M. A. H. Luttik, J. T. Pronk, E. J. Smid, P. A. Bron, P. Daran-Lapujade, “Transcriptome-Based Characterization of Interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus in Lactose-Grown Chemostat Cocultures”, Applied and Environmental Microbiology, vol. 79(19), pp. 5949-5961, 2013.
 S. Furukawa, T. Watanabe, H. Toyama, Y. Morinaga, “Significance of microbial symbiotic coexistence in traditional fermentation”, Journal of Bioscience and Bioengineering, vol. 116(5), pp. 533-539, 2013.
 G. M. Walker, G. G. Stewart, “Saccharomyces cerevisiae in the Production of Fermented Beverages”, Beverages, vol. 2(30), pp. 1-12, 2016.
 M.A. Calvo, E.L. Arosemena, C. Shiva, C. Adelantado, “Antimicrobial activity of plant natural extracts and essential oils (Science against microbial pathogens: communicating current research and technological advances, Formatex,)” , A. Mendes-Vilas, Ed. 2011, pp. 1179-1185.
 B. Cheirsilp, H. Shoji, H. Shimizu, S. Shioya, “Interactions between Lactobacillus keJiranofaciens and Saccharomyces cerevisiae in Mixed Culture for Kefiran Production”, Journal of Bioscience and Bioengineering, vol. 96(3), pp. 279-284, 2003.
 D. H. Kim, J. W. Chon, H. Kim, K. H. Seo, “Modulation of Intestinal Microbiota in Mice by Kefir Administration”, Food Sci. Biotechnol., vol. 24(4), 1397-1403, 2015.
 E. Ivanova, N. Atanasova-Pancevska, D. Kungulovski, “Antimicrobial Activities of Laboratory Produced Essential Oil Solutions Against Five Selected Fungal Strains”, Jour. Nat. Sci, Matica Srpska Novi Sad., vol. 124, 171-183, 2013.
 D. B. Makanjuola, D. G. Springham, “Identification of Lactic Acid Bacteria Isolated from Different Stages of Malt Whisky Distillery Fermentations”, J. Inst. Brew., vol. 90, pp. 13-19, 1984.
 C. Garofalo, G. Silvestri, L. Aquilanti, F. Clementi, “PCR-DGGE analysis of lactic acid bacteria and yeast dynamics during the production processes of three varieties of Panettone”, Journal of Applied Microbiology, vol. 105, pp. 243-254, 2008.
 İ. Çakır, “Antibacterial and antifungal activities of some lactic acid bacteria isolated from naturally fermented herbs”, Journal of Food, Agriculture & Environment, vol. 8(2): 223-226, 2010.
 S. M. Al-khazraji, “Preliminary screening of the antibacterial activity of Cinnamonum zeylanicum (cinnamon) barks”, IOSR Journal of Pharmacy and Biological Sciences, vol. 9(5), pp. 30-34, 2014.
 M. Balouiri, M. Sadiki, S. K. Ibnsouda, “Methods for in vitro evaluating antimicrobial activity: A review”, Journal of Pharmaceutical Analysis, vol. 6, pp. 71–79, 2016.
 J. Zaragoza, Z. Bendiks, C. Tyler, M. E. Kable, T. R. Williams, Y. Luchkovska, E. Chow, K. Boundy-Mills, M. L. Marco, “Effects of Exogenous Yeast and Bacteria on the Microbial Population Dynamics and Outcomes of Olive Fermentations”, Applied and Environmental Science, vol. 2(1), pp. 1-14, 2017.
 I. C. Gunsalus, C. F. Niven, “The Effect of pH on the Lactic Acid Fermentation”, J. Biol. Chem., vol. 145, pp. 131-136, 1942.
 K.H. Steinkraus, “Lactic Acid Fermentations (Chapter 5)”, Applications of Biotechnology in Traditional Fermented Foods, National Academy Press, Washington, D.C. pp. 43-51, 1992.
 M. A. Abdel-Rahman, Y. Tashiro, K. Sonomoto, “Recent advances in lactic acid production by microbial fermentation processes”, Biotechnology Advances, vol. 31, pp. 877–902, 2013.