Isolation and Molecular Characterization Of Antimicrobial Agent Producing Bacteria Isolated From Palandoken Mountain

Abstract

Streptomyces is a genus of Gram-positive bacteria that grows in different habitats, and its shape takes after filamentous fungi. The most effective characteristic of Streptomyces is the ability to produce secondary metabolites, such as antivirals, antifungals, anti-hypertensives, antitumorals, and especially antibiotics. In this study, bacteria producing antimicrobial substances were isolated from soil samples collected from Palandöken Mountain. Test strains were identified using conventional (morphological, physiological and biochemical tests) and molecular methods (16S rRNA sequencing). Then, the antagonistic effect of these bacteria against pathogenic microorganisms was determined by disc diffusion method. As a result of analysis, it was found that two bacteria (AO1 and AO3) were similar to Streptomyces violaceochromogenes, one (AO2) to Streptomyces ambofaciens and other (AO4) to Sphingomonas melonis at a rate of 99%. According to conventional tests, all isolates were catalase positive, three were oxidase negative (except AO4). In addition, pH, NaCl and temperature values that isolates can growth were determined. Finally, phylogenetic trees of the isolates were performed by the neighbor-joining method.

Keywords

• Antimicrobial subtances
• Streptomyces
• Molecular characterization
• Polyphasic approach

Palandöken Dağı’ndan İzole Edilen, Antimikrobiyal Ajan Üreten Bakterilerin İzolasyonu Ve Moleküler Karakterizasyonu

Öz

Streptomycesler, farklı habitatlarda gelişen bir Gram-pozitif bakteri cinsi olup, şekli ipliksi mantarlara benzer. Streptomyces'in en etkili özelliği antiviraller, antifungaller, anti-hipertansifler, antitümoraller ve özellikle antibiyotikler gibi sekonder metabolitler üretebilmesidir. Bu çalışmada, test suşları, geleneksel (morfolojik, fizyolojik ve biyokimyasal testler) ve moleküler yöntemler (16S rRNA sekanslama) kullanılarak tanımlandı. Daha sonra bu bakterilerin patojen mikroorganizmalara karşı antagonistik etkisi disk difüzyon yöntemi ile belirlendi. Analiz sonucunda, izole edilen dört bakteriden, ikisinin(AO1 ve AO3) Streptomyces violaceochromogenes'e, birinin (AO2) Streptomyces ambofaciens'e ve diğerinin ise (AO4) Sphingomonas melonis'e% 99 oranında benzer olduğu bulundu. Geleneksel testlere göre, tüm izolatlar katalaz pozitif, üçü oksidaz negatiftir (AO4 hariç). Ayrıca izolatların büyüyebileceği pH, NaCl ve sıcaklık değerleri belirlendi. Son olarak izolatların filogenetik ağaçları komşu birleştirme yöntemi ile yapılmıştır.

Anahtar Kelimeler

• Antimikrobiyal maddeler
• Streptomyces
• Moleküler karakterizasyon
• Polifazik yaklaşım

Kaynakça

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