Sucul çevreden izole edilen aktinomisetlerden biyoaktif bileşiklerin taranması, izolasyonu ve üretim ortamının optimizasyonu
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Dosyalar
Tarih
2018
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
Bu tez çalışmasında; aktinomiset üyesi olan mikroorganizmaların izolasyonu, antimikrobiyal aktivitelerin taranması ve üretim ortamının optimizasyonu gerçekleştirilmiştir. İzmir’in İnciraltı bölgesinde bulunan lagün sedimentinden 16 adet aktinomiset izolatı elde edilmiştir. Cross-streak (Cs) metodu kullanılarak birincil antimikrobiyal aktivite taranmıştır. Birincil antimikrobiyal aktivite tarama sonuçlarına göre, ERS13 izolatı en yüksek aktiviteyi göstermiştir. İzolatın fenotipik ve genotipik özelliklerine göre; beyazımtrak koloni tipinde, aerobik, sıvı ortamda pellet oluşturan, hareketsiz, filamentli, gram pozitif ve Streptomyces variabilis VITUMVB03 suşuna %99 oranında benzerlik gösterdiği belirlenmiştir. İkincil antimikrobiyal aktivite taranmasında disk difüzyon metodu kullanılmıştır. Tarama sonuçlarına göre ERS13 izolatının, Bacillus cereus ATCC 7064, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 29998, Salmonella typhimurium, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 6538/P, Aspergillus niger ve Candida albicans patojenlerine karşı sırasıyla 10 mm, 24 mm, 13 mm, 15 mm, 20 mm, 16 mm, 18 mm ve 12 mm inhibisyon zonu oluşturduğu gözlemlenmiştir. Üretim ortamı optimizasyonu ise ERS13 izolatının ekstrakte ürünün kullanıldığı disk difüzyon testi ile belirlenmiştir. Test sonuçlarına göre en aktif üretim ortamı; soya unu (20,0 g/l), glukoz (5,0 g/l), riboz (5,0 g/l), gliserol (5,0 g/l), patates nişastası (5,0 g/l), amonyum sülfat (0,2 g/l), maya ekstraktı (2,0 g/l) ve Bacto pepton (2,0 g/l) besiyeri bileşenlerine sahip Carbohydrate carcode medium (CCM) olarak belirlenmiştir.
In this thesis, the isolation of microorganisms which are the member of actinomycetes, the screening of antimicrobial activities and optimization of production medium has been carried out. 16 actinomycete isolates were obtained from lagoon sediment which is located in Inciralti, Izmir region. Primary antimicrobial activity was screened using the Cross-streak method. According to the primary antimicrobial activity screening results, the ERS13 isolate has showed the highest activity. According to phenotypic and genotypic characteristics of the isolate; it has been determined that it is whitish in colour, aerobic, pellet forming in broth culture, non motile, filamentous, gram positive and 99% similar to Streptomyces variabilis strain VITUMVB03. Disc diffusion method was used for secondary antimicrobial activity screening. The ERS13 isolate has showed the highest inhibition zone against Bacillus cereus ATCC 7064, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 29998, Salmonella typhimurium, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 6538/P, Aspergillus niger and Candida albicans with a zone of inhibition of 10 mm, 24 mm, 13 mm, 15 mm, 20 mm, 16 mm, 18 mm ve 12 mm, respectively. The production medium optimization was determined by disc diffusion test using extracts of the ERS13 isolate. According to the test results, Carbohydrate carcode medium (CCM) was determined as the most active production medium which contain soyaflour (20.0 g/l), glucose (5.0 g/l), ribose (5.0 g/l), glycerol (5.0 g/l), potato starch (5.0 g/l), ammonium sulphate (0.2 g/l), yeast extract (2.0 g/l) and Bactopeptone (2.0 g/l).
In this thesis, the isolation of microorganisms which are the member of actinomycetes, the screening of antimicrobial activities and optimization of production medium has been carried out. 16 actinomycete isolates were obtained from lagoon sediment which is located in Inciralti, Izmir region. Primary antimicrobial activity was screened using the Cross-streak method. According to the primary antimicrobial activity screening results, the ERS13 isolate has showed the highest activity. According to phenotypic and genotypic characteristics of the isolate; it has been determined that it is whitish in colour, aerobic, pellet forming in broth culture, non motile, filamentous, gram positive and 99% similar to Streptomyces variabilis strain VITUMVB03. Disc diffusion method was used for secondary antimicrobial activity screening. The ERS13 isolate has showed the highest inhibition zone against Bacillus cereus ATCC 7064, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 29998, Salmonella typhimurium, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 6538/P, Aspergillus niger and Candida albicans with a zone of inhibition of 10 mm, 24 mm, 13 mm, 15 mm, 20 mm, 16 mm, 18 mm ve 12 mm, respectively. The production medium optimization was determined by disc diffusion test using extracts of the ERS13 isolate. According to the test results, Carbohydrate carcode medium (CCM) was determined as the most active production medium which contain soyaflour (20.0 g/l), glucose (5.0 g/l), ribose (5.0 g/l), glycerol (5.0 g/l), potato starch (5.0 g/l), ammonium sulphate (0.2 g/l), yeast extract (2.0 g/l) and Bactopeptone (2.0 g/l).
Açıklama
Anahtar Kelimeler
Biyoaktif Bileşik, Ekstraksiyon, Sekonder Metabolit, Disk Difüzyon, Sucul Aktinomiset, Cross-Streak Metodu, Bioactive Compound, Extraction, Secondary Metabolite, Disc Diffusion, Aquatic Actinomycete, Cross-Streak Method