Doğu Karadeniz Bölgesindeki Karstik Mağaralardan İzole Edilen Aktinomisetlerin Antimikrobiyal Aktivitelerinin ve 16S rRNA Dizilerinin Araştırılması

Year 2023, Volume: 26 Issue: 6, 1277 - 1290, 31.12.2023

Enis Fuat Tüfekci Ümit Uzun Nagihan Sağlam Ertunga Ayşenur Biber İnanç Alptuğ Hıdıroğlu İlayda Tekkılıç Birhan Altay Ali Osman Kılıç

https://doi.org/10.18016/ksutarimdoga.vi.1226184

Abstract

Günümüzde kullanılan antibiyotiklerin büyük çoğunluğunun başta Streptomyces cinsi olmak üzere çeşitli aktinomiset grubu bakterilerden orijin aldığı düşünüldüğünde, mağara gibi ekstrem ortamlardan izole edilecek yeni aktinomisetlerin tıp dünyasına yeni antibiyotikler kazandırabileceği öngörülmektedir. Bu çalışmada, üç farklı karstik mağaradan izole edilen aktinomiset izolatlarının antimikrobiyal aktivitelerinin araştırılması ve etkili izolatların moleküler yöntemlerle tanımlanması amaçlanmıştır. Çalışmaya Türkiye’nin Doğu Karadeniz Bölgesinde bulunan Gümüşhane ilindeki Akçakale, Kırklar (Altıntaş) ve Köprübaşı mağaralarından izole edilen 179 aktinomiset izolatı dahil edilmiştir. İzolatların antimikrobiyal aktiviteleri yedi Gram-negatif, üç Gram-pozitif ve bir maya suşuna karşı çapraz çizgi yöntemi ile araştırılmıştır. Elli üç izolatın (%29,6) test edilen mikroorganizmalardan en az birine karşı antimikrobiyal aktiviteye sahip olduğu bulunmuştur. İzolatların Staphylococcus aureus, Bacillus subtilis, Enterococcus faecalis, Chromobacterium violaceum, Klebsiella pneumoniae, Salmonella Typhimurium, Escherichia coli, Acinetobacter haemolyticus, Pseudomonas aeruginosa, Enterobacter aerogenes ve Candida albicans’a karşı antimikrobiyal aktivite sergileme oranı sırasıyla %21.2 %20.0, %16.8, %12.8, %3.4, %2.8, %2.2, %1.1, %0.6, %0.6, %0.6 şeklinde bulunmuştur. TRMS 124 olarak adlandırılan bir aktinomiset izolatı, 10 test mikroorganizmasına karşı antimikrobiyal aktivite sergilemiştir. En az üç test mikroorganizmasına karşı antimikrobiyal aktivite sergileyen aktinomisetler arasından randomize olarak seçilen 26 izolatın tanımlanması ve filogenetik analizi için 16S ribozomal RNA (16S rRNA) dizi analizi yapılmıştır. Buna göre 24 izolatın çeşitli Streptomyces türleri ile iki izolatın ise sırasıyla Embleya scabrispora ve Couchioplanes caeruleus ile homoloji gösterdiği tespit edilmiştir. Bu çalışmadan elde edilen bulgular karstik mağaraların antimikrobiyal madde üretme potansiyeline sahip aktinomisetlerin izolasyonu için doğal kaynaklar olabileceğini göstermiştir.

Keywords

16S ribosomal RNA, Actinomycetes, Antimicrobial activity, Cave, Streptomyces

Supporting Institution

Karadeniz Teknik Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

FBA-2016-5481

Investigation of Antimicrobial Activities and 16S rRNA Sequences of Actinomycetes Isolated from Karst Caves in the Eastern Black Sea Region of Türkiye

Abstract

Considering that most antibiotics originate from actinomycete group bacteria, especially the Streptomyces genus, it is predicted that novel actinomycetes isolated from extreme environments such as caves may bring novel antibiotics to the medical world. The study aimed to screen the antimicrobial activity of actinomycetes isolated from the three karst caves in Türkiye and to identify selected isolates with antimicrobial activity by molecular methods. One hundred seventy-nine actinomycetes isolated from Akçakale, Kırklar (Altıntaş), and Köprübaşı Caves in Gümüşhane province in the Eastern Black Sea Region of Türkiye were included in the study. The antimicrobial activity of isolates was investigated using the modified cross-streak agar method against seven Gram-negative bacteria, three Gram-positive bacteria, and one yeast strain. Fifty-three isolates (29.6%) had antimicrobial activity against at least one of the tested microorganisms. The rate of isolates exhibiting antimicrobial activity against Staphylococcus aureus, Bacillus subtilis, Enterococcus faecalis, Chromobacterium violaceum, Klebsiella pneumoniae, Salmonella Typhimurium, Escherichia coli, Acinetobacter haemolyticus, Pseudomonas aeruginosa, Enterobacter aerogenes, and Candida albicans was 21.2%, 20.0%, 16.8%, 12.8%, 3.4%, 2.8%, 2.2%, 1.1%, 0.6%, 0.6%, and 0.6%, respectively. An actinomycete isolate, TRMS 124, showed antimicrobial activity against ten test microorganisms. The 16S ribosomal RNA (16S rRNA) sequencing was performed for the identification and phylogenetic analysis of 26 isolates randomly selected among actinomycetes that exhibited antimicrobial activity against at least three test microorganisms. As a result, it was determined that 24 isolates showed homology with various Streptomyces species and two isolates with Embleya scabrispora and Couchioplanes caeruleus, respectively. These results showed that karst caves could be good sources for isolating actinomycetes with the potential to produce antimicrobial compounds.

Keywords

16S ribosomal RNA, Actinomycetes, Antimicrobial activity, Cave, Streptomyces

Project Number

FBA-2016-5481

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