KEFİR ÖRNEKLERİNDEN LAKTİK ASİT BAKTERİLERİNİN İZOLASYONU, İDENTİFİKASYONU VE ANTİMİKROBİYAL ETKİLERİNİN DEĞERLENDİRİLMESİ
Year 2020,
, 72 - 80, 01.12.2019
Sevim Feyza Erdoğmuş
,
Barış Bostancı
Abstract
Bu çalışmada, Afyonkarahisar İli'nde satışa sunulan markasız, paketsiz kefir örneklerindeki laktik asit bakterilerinin izolasyonu, tanımlanması ve patojen test mikroorganizmaları üzerindeki antimikrobiyel etkinliği değerlendirilmiştir. 5 farklı kefir örneğinden 6 adet laktik asit bakterisi izole edilmiş ve izolatların moleküler tanımlanması için 16S rDNA PZR analizi yapılmıştır. İzolatların 16S rDNA sekans analiz sonuçlarına göre, izole edilen bakteriler; Lactobacillus fermentum, Enterococcus faecium veEnterococcus durans türleri olarak tanımlanmıştır. Laktik asit bakteri izolatlarının patojen test mikroorganizmaları üzerindeki antimikrobiyel aktivitesini belirlemek için agar spot ve agar kuyu difüzyon testi kullanılmıştır. Tüm izolatlar, test edilen patojen mikroorganizmalar üzerinde farklı seviyelerde antimikrobiyel etki göstermiştir. Laktik asit bakteri izolatları, Klebsiella pneumoniea NRRLB 4420, Pseudomonas aeuroginosa ATCC 11778, Listeria monocytogenes ATCC 1911 üzerinde diğer test mikroorganizmalarına (Staphylococcus aureus ATCC 25923, Streptococcus feacalis, Escherichia coli ATCC 35218, Bacillus subtilis NRS-744) göre daha yüksek antimikrobiyel etki göstermiştir..
Supporting Institution
Afyon Kocatepe Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi
Project Number
17.SAĞ.BİL.06
Thanks
Bu çalışma; Afyon Kocatepe Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından 17.SAĞ.BİL.06 numaralı proje ile desteklenmiştir.
References
- Abriouel, H., Gálvez, A., López, R.L., Omar, N.B. (2007). Bacteriocin based strategies for food biopreservation. Int J Food Microbiol, 120: 51-70.
- Amirbozorgi, G., Samadlouie, H., Shahidi, S.A. (2016). Identification and characterization of lactic acid bacteria ısolated from Iranian traditional dairy products, IBBJ Winter, 2 (1): 47-52.
- Arabestani, M.R., Mousavi, S.M., Alikhani, M.Y. (2014). Bacteriocins as the alternatives to antibiotics, Avicenna J Clin Microbiol Infect, doi: 10.17795/ajcmi-21086.
- Bayram, M., Yıldırım, Z. (2016). Beyaz peynirden bakteriyosin üreten bakterinin (Enterococcus faecium) izolasyonu ve bakteriyosinin karakterizayonu, Gaziosmanpaşa Bil Araş Derg, 13:103-115.
- Bennani, S, Mchiouer, K., Rokni, Y., Meziane, M. (2017). Characterisation and identification of lactic acid bacteria isolated from Moroccan raw cow’s milk. J Mater Environ Sci, 8(S), 4934-4944.
- Blaser, M. (2011). Antibiotic overuse: stop the killing of beneficial bacteria, Nature, 476 (7361): 393-394.
- Chen, C.C., Wang, S.Y., Chen, M.J. (2008)., Microbiological study of lactic acid bacteria in kefir grains by culture-dependent and culture-independent methods, Food Microbiol, 25 (3): 492-501.
- Cooper, M.A., Shlaes, D. (2011). Fix the antibiotics pipeline, Nature, 472 (7341): 32. doi: 10.1038/472032a
- Corr, S.C., Li, Y. Riedel, C.U., O’Toole, P.W., Hill, C., Gahan, C.G. (2007). Bacteriocin production as a mechanism for the antiinfective activity of Lactobacillus salivarius UCC118, Proceed Nat Acad Sci, 104: 7617-7621.
- De Martinis, E.C.P., Alves, V.F., Franco, B.D.G.M. (2002). Fundamentals and perspectives for the use of bacteriocins produced by lactic acid bacteria in meat. Food Rev Int, 18 (2): 191-208.
- Deegan, L.H., Cotter, P.D., Colin, H., Ross, P. (2006). Bacteriocins: biological tools for bio-preservation and shelf-life extension, Int Dairy J, 16: 1058-1071.
- Dinçer, E., Kıvanç, M., Karaca, H. (2010). Biyokoruyucu olarak laktik asit bakterileri, Gıda, 35(1): 1-8.
- Eijsink, V.G.H., Axelsson, L., Diep, D.B., Havarstein, L.S., Holo, H., Nes, I.F. (2002). Production of class II bacteriocins by LAB; an example of biological warfare and communication, Antonie van Leeuwenhoek, 81: 639-654.
- Erdoğrul, Ö.Z, Çetin, Ö., Ergün, Ö. (2002). Fermente sucuklardan izole edilen Pediococcus pentosaceus suşlarının bazı metabolik ve antimikrobiyal aktiviteleri üzerine çalışmalar. İstanbul Üni Vet Fak Derg, 28 (1): 249-254.
- Evren, M., Albayram, C., Apan, M. (2006), Laktik asit bakterilerinin oluşturduğu antimikrobiyal maddeler. Türkiye 9. Gıda Kongresi, Mayıs 24-26, Bolu, Türkiye, 977-980.
- Goh, H.F., Philip, K. (2015). Purification and characterization of bacteriocin produced by Weissella confusa A3 of dairy origin, PLOS ONE,| doi:10.1371/journal.pone.0140434.
- Gülgör, G., Özçelik, F. (2014), Bakteriyosin üreten laktik asit bakterilerinin probiyotik amaçlı kullanımı, Akademik Gıda, 12(1): 63-68.
- İşleroğlu, H., Yıldırım, Z., Yıldırım, M. (2008). Yöresel peynirden antimikrobiyal aktiviteye sahip laktik asit bakterisinin izolasyonu ve tanısı, Gazi Osmanpaşa Üni Ziraat Fak Derg, 25 (1): 1-6.
- Kaur, S. (2015). Bacteriocins as potential anticancer agents, Front Pharmacol. 6: 272, doi: 10.3389/fphar.2015.00272.
- López-Cuellar, R., Rodríguez-Hernández, A.I., Chavarría-Hernández, N. (2016). LAB bacteriocin applications in the last decade, Biotechnology & Biotechnological Equipment, 30(6): 1039-1050.
- Masalam, M. S, Bahieldin A., Alharbi M.G., Al-Masaudi, S., Al-Jaouni S.K., Harakeh, S.M., Al-Hindi R. (2018). Isolation, molecular characterization and probiotic potential of lactic acid bacteria in Saudi raw and fermented milk. Evid Based Complement Alternat Med, doi.org/10.1155/2018/7970463.
- Moshood, A.Y., Tengku, H.A. (2013). Lactic acid bacteria: Bacteriocin producer. IOSR J Pharmacy, 3(4): 44-50.
- Nespolo, C.R., Brandelli, A. (2010). Production of bacteriocin-like substances by lactic acid bacteria ısolated from regional ovine cheese, Brazillian J Microbiol, 41(4): 1009-1018.
- Pringsulaka, O., Thongngam, N., Suwannasai, N., Atthakor, W., Pothivejkul, K., Rangsırujı, A. (2012). Partial characterisation of bacteriocins produced by lactic acid bacteria isolated from thai fermented meat and fish. Food Cont, 23: 547- 551.
- Rodrıguez, E.G.B., Gaya, P., Nanez, M., Medina, M. (2000). Diversity of bacteriocins produced by lactic acid bacteria ısolated from raw milk, Int Dairy J , 10: 7-15.
- Schnurer, J., Magnusson, J. (2005), Antifungal activity against Aspergillus parasiticus of supernatants from whey permeates fermented with kefir grains, antifungal lactic acid bacteria as biopreservatives. Trend Food Sci Technol, 16: 70-78.
- Simova, E., Beshkova, D., Angelov, A., Hristozova, T., Frengova, G., Spasov, Z. (2002), Lactic acid bacteria and yeasts in kefir grains and kefir made from them. J Ind Microbiol Biotechnol., 28(1):1-6.
- Turner, P.C., Mclennan, A.G., Bates, A.D., White, M.R.H. (2004). Moleküler Biyoloji Önemli Notlar, Konuk, M. (Eds) Nobel Yayın, 613: 346 s.
- Whıte, A.R. (2011). Bsac working party on the urgent need: Regenerating antibacterial drug discovery, development. effective antibacterials: at what cost? the economics of antibacterial resistance and its control, J Antimicrob Chemother, 66(9):1948–1953.
- Yang, S.C., Lin, C.H., Sung, C.T., Fang, J.Y. (2014). Antibacterial activities of bacteriocins: Application in foods and pharmaceuticals, Front Microbiol, doi: 10.3389/fmicb.2014.00241, 2014.
- Zhang, Z., Vriesekoop, F., Yuan, Q., Liang, H. (2014). Effects of nisin on the antimicrobial activity of d-limonene and its nanoemulsion, Food Chem, 150:307-312.Wassie, M., Wassie T. (2016). Isolation and identification of lactic acid bacteria from raw cow milk. Int J Adv Res Biol Sci,. 3(8): 44-49.
ISOLATION AND IDENTIFICATION OF LACTIC ACID BACTERIA FROM KEFIR SAMPLES AND EVALUATION OF THEIR ANTIMICROBIAL EFFECTS
Year 2020,
, 72 - 80, 01.12.2019
Sevim Feyza Erdoğmuş
,
Barış Bostancı
Abstract
In this study, lactic acid bacteria were isolated from
unmarked, unpacked kefir samples that sold in Afyonkarahisar province, and
their antimicrobial activity on pathogen test microorganisms were evaluated. 6
lactic acid bacteria were isolated from 5 different kefir samples and 16S rDNA
PCR analysis was performed for molecular identifications of these isolates.
According to results of 16S rDNA PCR analysis; isolated bacteria from kefir
samples were identifed as Lactobacillus fermentum, Enterococcus
faecium and Enterococcus durans. Agar spot and agar-well diffusion
assay methods were used to determine the antimicrobial activity of the lactic
acid bacteria isolates on pathogen test microorganisms. All of the isolates
showed antimicrobial activity at different levels on the tested pathogenic
microorganisms. Lactic acid bacteria isolates showed higher antimicrobial
activity on Klebsiella pneumoniea NRRLB 4420, Pseudomonas aeuroginosa
ATCC 11778, Listeria monocytogenes ATCC 1911 than other test
microorganisms (Staphylococcus aureus ATCC 25923,
Streptococcus feacalis, Escherichia coli ATCC 35218, Bacillus
subtilis NRS-744).
Project Number
17.SAĞ.BİL.06
References
- Abriouel, H., Gálvez, A., López, R.L., Omar, N.B. (2007). Bacteriocin based strategies for food biopreservation. Int J Food Microbiol, 120: 51-70.
- Amirbozorgi, G., Samadlouie, H., Shahidi, S.A. (2016). Identification and characterization of lactic acid bacteria ısolated from Iranian traditional dairy products, IBBJ Winter, 2 (1): 47-52.
- Arabestani, M.R., Mousavi, S.M., Alikhani, M.Y. (2014). Bacteriocins as the alternatives to antibiotics, Avicenna J Clin Microbiol Infect, doi: 10.17795/ajcmi-21086.
- Bayram, M., Yıldırım, Z. (2016). Beyaz peynirden bakteriyosin üreten bakterinin (Enterococcus faecium) izolasyonu ve bakteriyosinin karakterizayonu, Gaziosmanpaşa Bil Araş Derg, 13:103-115.
- Bennani, S, Mchiouer, K., Rokni, Y., Meziane, M. (2017). Characterisation and identification of lactic acid bacteria isolated from Moroccan raw cow’s milk. J Mater Environ Sci, 8(S), 4934-4944.
- Blaser, M. (2011). Antibiotic overuse: stop the killing of beneficial bacteria, Nature, 476 (7361): 393-394.
- Chen, C.C., Wang, S.Y., Chen, M.J. (2008)., Microbiological study of lactic acid bacteria in kefir grains by culture-dependent and culture-independent methods, Food Microbiol, 25 (3): 492-501.
- Cooper, M.A., Shlaes, D. (2011). Fix the antibiotics pipeline, Nature, 472 (7341): 32. doi: 10.1038/472032a
- Corr, S.C., Li, Y. Riedel, C.U., O’Toole, P.W., Hill, C., Gahan, C.G. (2007). Bacteriocin production as a mechanism for the antiinfective activity of Lactobacillus salivarius UCC118, Proceed Nat Acad Sci, 104: 7617-7621.
- De Martinis, E.C.P., Alves, V.F., Franco, B.D.G.M. (2002). Fundamentals and perspectives for the use of bacteriocins produced by lactic acid bacteria in meat. Food Rev Int, 18 (2): 191-208.
- Deegan, L.H., Cotter, P.D., Colin, H., Ross, P. (2006). Bacteriocins: biological tools for bio-preservation and shelf-life extension, Int Dairy J, 16: 1058-1071.
- Dinçer, E., Kıvanç, M., Karaca, H. (2010). Biyokoruyucu olarak laktik asit bakterileri, Gıda, 35(1): 1-8.
- Eijsink, V.G.H., Axelsson, L., Diep, D.B., Havarstein, L.S., Holo, H., Nes, I.F. (2002). Production of class II bacteriocins by LAB; an example of biological warfare and communication, Antonie van Leeuwenhoek, 81: 639-654.
- Erdoğrul, Ö.Z, Çetin, Ö., Ergün, Ö. (2002). Fermente sucuklardan izole edilen Pediococcus pentosaceus suşlarının bazı metabolik ve antimikrobiyal aktiviteleri üzerine çalışmalar. İstanbul Üni Vet Fak Derg, 28 (1): 249-254.
- Evren, M., Albayram, C., Apan, M. (2006), Laktik asit bakterilerinin oluşturduğu antimikrobiyal maddeler. Türkiye 9. Gıda Kongresi, Mayıs 24-26, Bolu, Türkiye, 977-980.
- Goh, H.F., Philip, K. (2015). Purification and characterization of bacteriocin produced by Weissella confusa A3 of dairy origin, PLOS ONE,| doi:10.1371/journal.pone.0140434.
- Gülgör, G., Özçelik, F. (2014), Bakteriyosin üreten laktik asit bakterilerinin probiyotik amaçlı kullanımı, Akademik Gıda, 12(1): 63-68.
- İşleroğlu, H., Yıldırım, Z., Yıldırım, M. (2008). Yöresel peynirden antimikrobiyal aktiviteye sahip laktik asit bakterisinin izolasyonu ve tanısı, Gazi Osmanpaşa Üni Ziraat Fak Derg, 25 (1): 1-6.
- Kaur, S. (2015). Bacteriocins as potential anticancer agents, Front Pharmacol. 6: 272, doi: 10.3389/fphar.2015.00272.
- López-Cuellar, R., Rodríguez-Hernández, A.I., Chavarría-Hernández, N. (2016). LAB bacteriocin applications in the last decade, Biotechnology & Biotechnological Equipment, 30(6): 1039-1050.
- Masalam, M. S, Bahieldin A., Alharbi M.G., Al-Masaudi, S., Al-Jaouni S.K., Harakeh, S.M., Al-Hindi R. (2018). Isolation, molecular characterization and probiotic potential of lactic acid bacteria in Saudi raw and fermented milk. Evid Based Complement Alternat Med, doi.org/10.1155/2018/7970463.
- Moshood, A.Y., Tengku, H.A. (2013). Lactic acid bacteria: Bacteriocin producer. IOSR J Pharmacy, 3(4): 44-50.
- Nespolo, C.R., Brandelli, A. (2010). Production of bacteriocin-like substances by lactic acid bacteria ısolated from regional ovine cheese, Brazillian J Microbiol, 41(4): 1009-1018.
- Pringsulaka, O., Thongngam, N., Suwannasai, N., Atthakor, W., Pothivejkul, K., Rangsırujı, A. (2012). Partial characterisation of bacteriocins produced by lactic acid bacteria isolated from thai fermented meat and fish. Food Cont, 23: 547- 551.
- Rodrıguez, E.G.B., Gaya, P., Nanez, M., Medina, M. (2000). Diversity of bacteriocins produced by lactic acid bacteria ısolated from raw milk, Int Dairy J , 10: 7-15.
- Schnurer, J., Magnusson, J. (2005), Antifungal activity against Aspergillus parasiticus of supernatants from whey permeates fermented with kefir grains, antifungal lactic acid bacteria as biopreservatives. Trend Food Sci Technol, 16: 70-78.
- Simova, E., Beshkova, D., Angelov, A., Hristozova, T., Frengova, G., Spasov, Z. (2002), Lactic acid bacteria and yeasts in kefir grains and kefir made from them. J Ind Microbiol Biotechnol., 28(1):1-6.
- Turner, P.C., Mclennan, A.G., Bates, A.D., White, M.R.H. (2004). Moleküler Biyoloji Önemli Notlar, Konuk, M. (Eds) Nobel Yayın, 613: 346 s.
- Whıte, A.R. (2011). Bsac working party on the urgent need: Regenerating antibacterial drug discovery, development. effective antibacterials: at what cost? the economics of antibacterial resistance and its control, J Antimicrob Chemother, 66(9):1948–1953.
- Yang, S.C., Lin, C.H., Sung, C.T., Fang, J.Y. (2014). Antibacterial activities of bacteriocins: Application in foods and pharmaceuticals, Front Microbiol, doi: 10.3389/fmicb.2014.00241, 2014.
- Zhang, Z., Vriesekoop, F., Yuan, Q., Liang, H. (2014). Effects of nisin on the antimicrobial activity of d-limonene and its nanoemulsion, Food Chem, 150:307-312.Wassie, M., Wassie T. (2016). Isolation and identification of lactic acid bacteria from raw cow milk. Int J Adv Res Biol Sci,. 3(8): 44-49.