Remzi CHOLAKOV, Zapryana DENKOVA, Velichka YANAKİEVA, Yulian TUMBARSKİ, Zoltan URSHEV, Iliyan DOBREV

Bozadan İzole Edilen Leuconostoc lactis BT17 Suşunun Biyokimyasal ve Moleküler-Genetik Yöntemlerle Tanımlanması

Balkanlar bölgesinde geleneksel bir doğal fermente tahıl ürünü olan bozadan, BT17 suşu izole edilmiştir. Katı MRS agar besiyerine çizim ve Gram boyama yöntemiyle koloni ve hücre morfolojisi belirlenmiştir. Gram pozitif olarak tespit edilen izolatın biyokimyasal tanımlaması API 50 CHL ve Apiweb®, moleküler-genetik yöntemlerden ise ARDRA çoğaltılmış rDNA restriksiyon analizi, uç endonükleaz enzimi EcoR I, Hae III ve Alu I kullanılarak, endonüklezlardan elde edilen restriksiyon profillerinin her bir suş için spesifik olduğu analiz yöntemi ve 16S rRNA dizi sekansı kullanılarak gerçekleştirilmiştir. Biyokimyasal tanımlamadan elde edilen sonuçlarda, izole edilen BT17 suşunun %59.1 Lactobacillus plantarum 1 ve %41.2 Lactobacillus pentosus suşuna ait olduğu tespit edilmiştir. Biyokimyasal tanımlama yöntemleri çabuk sonuç vermekte, fakat bazı farklılıklar göstermektedir. 16S rRNA dizi sekansına göre elde edilen sonuçlar izole edilen BT17 suşunu %99 oranında Leuconostoc lactis olarak tanımlarken, ARDRA yönteminden elde edilen sonuçlar izole edilen suşun restriksiyon profillerinin referans suşlarının Lactobacillus acidophilus DSM 20079, Lactobacillus helveticus DSM 20075, Lactobacillus casei ssp. casei DSM 20011, Lactobacillus plantarum DSM 20174, Lactobacillus delbrueckii ssp. bulgaricus DSM 20081 profilleriyle eşleşmediği tespit edilmiştir

Anahtar Kelimeler:

Leuconostoc, Tahıl, Boza, Sekans, API 50 CHL

Biochemical and Molecular Identification of a Strain Leuconostoc lactis BT17, Isolated from a Spontaneously Fermented Cereal Beverage, Boza

The strain BT17 was isolated from a naturally fermented cereal beverage boza , which is traditional in Balkan countries. The colony characteristics and cell morphology of the strain were determined after its cultivation on a solid culture medium and coloring of microscopic preparations. The strain was identified by biochemical methods - API 50 CHL kit system, followed by apiweb software processing, as well as molecular techniques amplified rDNA restriction analysis ARDRA with the endonucleases Eco RI, Hae III and Alu I, and nucleotide sequencing of the 16S rRNA gene. The results obtained from the biochemical identification showed that the isolated strain of BT17 is related to the species Lactobacillus plantarum 1 the rate of reliability with 59.1% and Lactobacillus pentosus the rate of reliability with 41.2% . The comparative 16S rRNA gene sequence-based phylogenetic analysis revealed 99% pairwise similarity of the studied strain to the bacterial species Leuconostoc lactis. The aim of this study was the identification, biochemical and molecular characterization of a strain Leuconostoc lactis BT17, isolated from a spontaneously fermented cereal beverage boza

Keywords:

Leuconostoc, Cereal, Boza, Sequencing, API 50 CHL,

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Tanguler, H., 2014. Traditional Turkish fermented cereal based products: Tarhana, boza and chickpea bread. Turkish Journal of Agriculture - Food Science and Technology 2(3): 144-149.
Zorba, M., Hancioglu, O., Genç, M., Karapinar, M., Ova, G., 2003. The use of starter cultures in the fermentation of Boza, a traditional Turkish beverage. Process Biochemistry 38: 140–141.
Moncheva, P., Chipeva, V., Kujumdzieva, A., Ivanova, I., Dousset, X., Gocheva, B., 2003. The composition of the microflora of boza, an original Bulgarian Biotechnology 17(1): 164-168. Biotechnology and
Arici, M., Daglioglu, O., 2002. Boza: a lactic acid fermented cereal beverage as a traditional Turkish food. Food Research International 18: 39-48.
Botes, A., Todorov, S.D., Von Mollendorff, J.W., Botha, A., Dicks, L.M.T., 2007. Identification of lactic acid bacteria and yeast from boza. Process Biochemistry 42: 267-270.
Caputo, L., Quintieri, L., Baruzzi, F., Borcakli, M., Morea, M., 2012. Molecular and phenotypic characterization of Pichia fermentans strains found among Boza yeasts. Food Research International 48: 755-762.
Hancioğlu, O., Karapinar, M., 1997. Microflora of Boza, a traditional fermented Turkish beverage. International Journal of Food Microbiology 35: 271- 274.
Todorov, S.D., Dicks, L.M.T., 2006. Screening for bacteriocin-producing lactic acid bacteria from boza, a traditional cereal beverage from Bulgaria. Comparison Biochemistry 41: 11-19. bacteriocins. Process
Delley, M., Mollet, B., Hottinger, H., 1990. DNA probe for Lactobacillus delbrueckii. Applied and Environmental Microbiology 56: 1967-1970.
Lane, D.J., 1991. 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systematics, Edited by E. Stackebrandt and M. Goodfellow, John Wiley and Sons, New York, NY, 115- 75p.
Dicks, L.M.T., Dellaglio, F., Collins, M.D., 1995. Proposal to reclassify Leuconostoc oenos as Oenococcus oeni [corrig.] gen. nov. comb. nov. International Journal of Systematic Bacteriology 45(2): 395-397.
Kim, D.S., Fogler, H.S., 1999. The effects of exopolymers on cell morphology and culturability of Leuconostoc mesenteroides during starvation. Applied Microbiology and Biotechnology 52: 839- 844.
Lee, H.J., Park, S.Y., Kim, J., 2000. Multiplex PCR- based detection and identification of Leuconostoc species. FEMS Microbiology Letters 193: 243-247.