Determination of probiotic and some technological properties of lactic acid bacteria isolated from cheeses sold in the Kilis region, Turkey

Abstract

Aims: The purpose of this study is to evaluate technological properties and probiotics potential of lactic acid bacteria (LAB) isolated from 15 raw milk Turkish Beyaz fresh cheese made in the Kilis region, Turkey.

Methods and Results: 287 colonies from 15 cheese samples were selected and 91 of them were analyzed as Gram-positive and catalase-negative. 19 strains of 91 colonies were accepted as potential probiotics since they stayed alive in pH 3.5 and 0.3% bile salt. They were identified as Lactobacillus sp. (11 strains), Leuconostoc sp. (3 strains), Streptococcus sp. (2 strains), Lactococcus sp. (2 strains), and Enterococcus sp. (1 strain) by biochemical tests and API test kit. It was determined that 19 strains with high acidification rate, 15 strains demonstrated weak proteolytic activity, 3 ones moderate, and 1 strong proteolytic activity. It was determined that 1 of the strains were resistant to chloramphenicol, 2 to tetracycline, and 6 to vancomycin. No strains resistant to penicillin and erythromycin could be detected. In the artificial gastric juice resistance test, although no viability was detected at pH 2.0. But it was determined that the viability values varied between 89.49-111.79% at pH 3.0. Also, these strains showed growth at bile salt. According to plasmid profiles of strains, 13 of 19 potential probiotic strains were determined to have plasmid DNA in the range of 1-5, while other strains were determined to not have plasmid DNA. The molecular sizes of the plasmid DNA of the isolated strains ranged from 2-16 kb.

Conclusions: In the research, it was determined that LABs isolated from Turkish white fresh cheese produced in Kilis have probiotic potential and can be used as starters in various fermented foods.

Significance and Impact of the Study: According to the results obtained, it was concluded that some strains can be used as probiotic starter culture in cheese production and others can be used for ripening of cheese.

Keywords

• Lactic acid bacteria
• starter culture
• technological properties
• probiotic properties
• Turkish beyaz fresh cheese

Kilis bölgesinde satılan peynirlerden izole edilen laktik asit bakterilerinin probiyotik ve bazı teknolojik özelliklerinin belirlenmesi

Öz

Amaç: Bu çalışmanın amacı Kilis yöresinde çiğ sütten üretilen 15 adet Türk Beyaz taze peynirinden izole edilen laktik asit bakterilerinin (LAB) teknolojik özellikleri ve probiyotik potansiyellerini değerlendirmektir.

Yöntem ve Bulgular: 15 peynir örneğinden 287 koloni seçilmiş ve bunlardan 91'i Gram pozitif ve katalaz negatif olarak analiz edilmiştir. 91 koloninin 19 suşu pH 3.5 ve %0.3 safra tuzunda canlı kaldıkları için potansiyel probiyotik olarak kabul edilmiştir. Biyokimyasal testler ve API test kiti kullanılarak yapılan tanımlama sonucunda Lactobacillus sp. (11 suş), Leuconostoc sp. (3 suş), Streptococcus sp. (2 suş), Lactococcus sp. (2 suş) ve Enterococcus sp. (1 suş) belirlenmiştir. Asidifikasyon hızı bakımından 19 suşun yüksek; proteolitik olarak 15 suşun zayıf, 3'ünün orta ve 1'inin güçlü aktivite gösterdiği belirlenmiştir. Antibiyotik dirençlilik açısından bakıldığında, suşlardan 1'inin kloramfenikole, 2'sinin tetrasikline ve 6'sının vankomisine dirençli olduğu belirlenmiştir. Penisilin ve eritromisine dirençli suş tespit edilememiştir. Yapay mide özsuyu direnç testinde pH 2,0'da canlılık saptanmamasına rağmen, pH 3.0'da canlılık değerlerinin %89,49-111,79 arasında değiştiği tespit edilmiştir. Suşların plazmit profillerine göre 19 potansiyel probiyotik suşun 13'ünün 1-5 aralığında plazmit DNA'sına sahip olduğu, diğer suşların plazmit DNA'sına sahip olmadığı belirlendi. İzole edilen suşların plazmit DNA'sının moleküler boyutları 2-16 kb arasında değişmekteydi.

Genel Yorum: Araştırmada, Kilis’te üretilen Türk beyaz taze peynirlerden izole edilen LAB’ların probiyotik potansiyelleri olduğu ve çeşitli fermente gıdalarda starter olarak da kullanılabilecekleri belirlenmiştir.

Çalışmanın Önemi ve Etkisi: Elde edilen sonuçlara göre bazı suşların peynir üretiminde probiyotik starter kültür olarak, bazılarının ise peynirin olgunlaşmasında kullanılabileceği sonucuna varılmıştır.

Anahtar Kelimeler

• Laktik asit bakterileri
• başlatıcı kültür
• teknolojik özellikler
• probiyotik özellikler
• Türk beyaz taze peynir

Kaynakça

1. Ammor MS, Florez AB, van Hoek AHAM, de los Reyes-Gavilan CG et al (2008) Molecular Characterization of Intrinsic and Acquired Antibiotic Resistance in Lactic Acid Bacteria and Bifidobacteria. J Mol Microbiol Biotechnol 14, 6–15.
2. Atiles MW, Dudley EG, Steele JL (2000) Gene Cloning, Sequencing, and Inactivation of the Branched-Chain Aminotransferase of Lactococcus lactis Lm0230. Appl Environ Microbiol 66(6), 2325-2329.
3. Başyiğit G (2004) The probiotic properties of some lactic acid bacteria. Master’s Thesis, Suleyman Demirel Univesity, Research Project Grants SDU-BAP-517.
4. Başyiğit Kılıç G, Kuleaşan H, Eralp I, Karahan AG (2009). Manufacture of Turkish Beyaz cheese added with probiotic strains. LWT-Food Science and Technology 42, 1003–1008.
5. Beyatlı Y, Tunail N (1982) Yoğurtlardan izole edilen kimi bakterilerin starter olarak seçilme olanakları üzerine bir araştırma. Tübitak TOVAG-414.
6. Bradley RL, Arnold E, Barbano DM, Semerad RG, Smith DE, Vines BK (1992) Chemical and Physical Methods. In: Marshall, T.(Ed). Standart Methods for The Examination of Dairy Products. American Public Healt Association, Washington Dc, Pp, 433-531.
7. Bulut Ç (2003) Isolation and molecular characterization of lactic acid bacteria from cheese. Master’s Thesis İzmir Teknoloji Enstitüsü 102s, İzmir.
8. Citti JE, Sandine WE, Elliker PR (1963) Some Observation on the Hull Method for Measurement of Proteolysis in Milk. Journal of Dairy Science 46, 337-345.

9. Cogan BTM, Barbosa M, Beuvier E, Bianchi-Salvadori B et al. (1997) Characterization of The Lactic Acid Bacteria in Artisanal Dairy Products. Journal of Dairy Research 64, 409-421.
10. Danielsen M, Wind A (2003) Susceptibility of Lactobacillus spp. to antimicrobial agents. International Journal of Food Microbiology, 82, 1-11.
11. Dikbaş Yıldız K (2013) Balıkesir yöresine ait peynirlerden izole edilen Lactobacillus bakterilerinin starter ve probiyotik özelliklerinin araştırılması. Gazi Üniversitesi, Fen Bilimleri Enstitüsü, Biyoloji Anabilim Dalı.
12. Dunne C, Murphy L, Flynn S, O’Mahony L et al. (1999) Probiotics: From myth to reality. Demonstration of functionality in animal models of disease and in human clinical trials. Antonie van Leeuwenhoek, 76, 279–292.
13. Egervarn M, Roos S, Lindmark H (2009) Identification and characterization of antibiotic resistance genes in Lactobacillus reuteri and Lactobacillus plantarum. Journal of Applied Microbiology 107, 1658–1668.
14. Ertekin Ö, Çon AH (2014) Industrial and Probiotic Characteristics of Lactic Acid Bacteria Isolated from Fermented Foods. Academic Food Journal 12(4), 6-16.
15. Ertürkmen P, Öner Z (2015) Beyaz Peynir Örneklerinden İzole Edilen Laktik Asit Bakterilerinin Başlatıcı (Starter) Kültür Özelliklerinin Biyokimyasal Yöntemlerle Belirlenmesi. Süleyman Demirel Üniversitesi, Fen Bilimleri Enstitüsü Dergisi, Cilt 19, Sayı 3, 9-16.
16. Franz CMAP, Specht I, Haberer P, Holzapfel WH (2001) Bile salt hydrolase activity of enterococci isolated from food: screening and quantitative determination. Journal of Food Protection 46, 725–729.
17. Gardiner G, Stanton C, Lynch PB, Collins JK et al. (1999) Evaluation of Cheddar cheese as a food carrier for delivery of a probiotic strain to the gastrointestinal tract. Journal Dairy Science, 82: 1379-1387.
18. Gonzalez CJ, Encinas JP, Garcia-Lopez ML, Otero A (2000) Characterization and identification of lactic acid bacteria from freshwater fishes. Food Microbiology, 17: 383–391.
19. Herreros MA, Fresno JM, Rieto MJ, Tornadijo ME (2003) Technological characterization of lactic acid bacteria isolated from Armada cheese (a Spanish goats‟ milk cheese). International Dairy Journal, 13, 469-479.
20. Hwanhlem N, Watthanasakphuban N, Riebroy S, Benjakul S et al. (2010) “Probiotic lactic acid bacteria from Kung-Som: isolation, screening, inhibition of pathogenic bacteria”, Int. J. Food Sci. and Technol. 45: 594-601.
21. Kandler O, Weiss N (1986) Genus Lactobacillus. Pages 1209–1234 in Bergey’s Manual of Systematic Bacteriology. Vol. 2. P. H. A. Sneath, N. S. Mair, M. E. Sharpe, and J. G. Holt, ed. Williams & Wilkins, Baltimore, MD.
22. Karahan AG, Başyiğit Kılıç G, Kart A, Şanlıdere Aloğlu H, Öner Z, Aydemir S, Erkuş O, Harsa Ş (2010) Genotypic identification of some lactic acid bacteria by amplified fragment length polymorphism analysis and investigation of their potential usage as starter culture combinations in Beyaz cheese manufacture. J. Dairy Sci. 93:1–11.
23. Karakuş M (1994) Beyaz Peynirlerden İzole Edilen Laktik Asit Bakterilerinin Asit Oluşturma ve Proteolitik Aktiviteleri. Gıda Dergisi, 19(4); 237-241.
24. Karasu N (2006) Antagonistic and probiotic lactic starter cultures from pickle and olive. Pamukkale University, M. Sc. Thesis in Food Engineering, Denizli. Kaur S, Das M (2011) Functional Foods: An Overview. Food Sci. Biotechnol 20(4): 861-875.
25. Lee Y, Salminen S (1995) The coming of age of probiotics. Trends in Food Science and Technology, 6, 241–245.
26. Mishra V, Prasad DN (2005) “Application of in vitro methods for selection of Lactobacillus casei strains as potential probiotics”. Int. J. Food Microbiol 103: 109-115.
27. Ouadghiri M, Amar M, Vancanneyt M, Swings J (2005) Biodiversity of lactic acid bacteria in Moroccan soft white cheese (Jben), FEMS Microbiology Letters, 251, 267-271.
28. Öner Z, Karahan AG, Şanlıdere H, Kılıc G, Gundoğdu A (2006) Selection of starter culture for Beyaz cheese. Page 46 in Proc. 1st International Egyptian-Jordanian Conference on Biotechnology and Sustainable Development: Current Status and Future Scenarios. Dec. 11–14, 2006. National Research Center, Cairo, Egypt.
29. Pouwels PH, Leer RJ (1993) Genetics of Lactobacilli: plasmids and gene expression. Antonie van Leeuwenhoek 64, 85–107.
30. Rossi F, Capodaglio A, Dellaglio F (2008) Genetic Modification of Lactobacillus plantarum by heterologous Gene Integration in a not Functional Region of the Chromosome. Appl Microbiol Biotechnol 80, 79–86.
31. Sağlam H, Karahan AG (2017) Plasmids in Lactic Acid Bacteria and Their Properties. Kafkas University, Institute of Natural and Applied Science Journal, Volume 10, Issue 2, 243-276.
32. Sağlam H (2013) Plasmid profiles of the identified strains of Lactobacillus plantarum and determination of their some properties. Ph.D. Thesis, Suleyman Demirel Univesity, Graduate School of Applied and Natural Sciences Department of Food Enginering. Isparta.
33. Schillinger U, Lucke FK (1987) Identification of Lactobacilli from Meat and Meat Products. Food Microbiology, 4: 199–208.
34. Smithies O (1955) Zone Electrophoressis in Starch Gels: Group Variations in the Serum Proteins of Normal Human Adults. Biochemical of Journal, 61, 629–641.
35. Turhan İ, Öner Z (2014) Determination of starter culture properties of lactic acid bacteria isolated from cheese. Gıda, 39(1), 9-15.
36. Vinderola CG, Reinheimer JA. Lactic acid starter and probiotic bacteria:a comperative “in vitro” study of probiotic characteristics and biological barrier resistance. Food Research Int. 2003, 36:895-904.
37. Yerlikaya O. Molecular and phenothpic medhods for lactic acid bacteria identification. Journal of Food and Feed Science-Technology 2014, 14:8-22.