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Bu çalışmanın amacı, Van Gölü’nden toplanan su ve sediment numunelerinden lakkaz ve mannanaz enzimi üretme kabiliyetlerine sahip bakterilerin karakterizasyonudur. Bu araştırmanın materyalini Van Gölü’nden toplanan su ve sediment numuneleri oluşturmuştur. İzolasyon çalışması dilüsyon plak yöntemi ile alkali besiyerinde gerçekleştirilmiştir. İki farklı koloni morfolojisine sahip izolat gözlemlenmiştir. İzolatlara koloni çapı, pigmentasyon, hücre şekli, gram reaksiyonu, indol testi, sitrat testi, voges-proskauer (vp), arabinoz, mannitol, sükroz, adonitol,metil_d-glikozit, selobiyoz, mannoz, salicin, traheloz, galaktoz, inositol, rafinoz, inulin, sorbitol, ksiloz, melesitoz, glikoz, arjinin, nitrat redüksiyonu, beta galaktosidaz, oksidaz, lakkaz enzim aktivitesi, mannanaz enzim aktivitesi, nişasta hidrolizi, tween 80 hidrolizi, skim milk hidrolizi ve ksilan hidrolizi testleri uygulanmıştır. İzolatların lakkaz enzimini üretme kabiliyetlerine %0.2 mM ABTS ve %0.1 mM CuSO4 ile bakılmıştır. Mannanaz enzimi üretme kabiliyetleri %1 mannoz ve fenol içeren alkali besiyeri kullanılarak belirlenmiştir. Genomik DNA’sı izole edilen 2 izolatın 16S rRNA gen bölgesi 27F ve 1492R evrensel primerleri ile amplifiye edilip sekans analizi yapılmıştır. Mega 7.0.18 paket programı ile Maksimum Olabilirlik algoritması seçilerek Jukes ve Cantor'un uzaklık matrisi ile filogenik ağaçlar oluşturulmuştur. İzolatların tümünün lakkaz enzimi yönünden negatif olduğu, mannanaz enzimi yönünden N1 ve N2 izolatlarının pozitif olduğu görülmüştür. N1 izolatı için oluşturulan filogenetik ağaçta Halomonas bakterileri kendi aralarında güçlü bir homoloji ile kümelenmiştir. N2 izolatı için oluşturulan filogenetik ağaçta Bacillus bakterileri genel olarak güçlü bir homoloji ile kümelenmiştir. N1 ve N2 izolatları, morfolojik, fizyolojik, biyokimyasal özelliklerine ve 16S rRNA gen bölgesi analizine göre N1 Halomonas stevensii ve N2 Bacillus agaradhaerens olarak teşhis edilmiştir.
The purpose of this study is to characterize bacteria with the ability to produce lakacus and mananaz enzymes from the water and sediment samples collected from the Van Lake. The material of this research was formed by water and sediment samples collected from the Van Lake. The isolation work was carried out in alkaline foods with the dilusion plate method. It has been observed with two different colonial morphologies. Isolates have been applied colonial diameter, pigmentation, cell shape, gram reaction, indol test, citrate test, voges-proskauer (vp), arabinoz, mannitol, sulcose, adonitol,methyl_d-glycosite, selobioz, mannosis, salicin, traheloz, galactose, inositol, rafinoz, inulin, sorbitol, xylos, melesitoz, glucose, arginine, nitrate reduction, beta galactosidase, oxidase, lakacase enzyme activity, mananaz enzyme activity, niche hydrolysis, tween 80 hydrolysis, skim milk hydrolysis and hydrolysis tests. The capabilities of the insulates to produce lakacus enzyme were assessed by 0.2% mM ABTS and 0.1% mM CuSO4. Mananaz enzyme production capacity is determined by the use of 1% mannose and phenol-containing alkali foods. Genetic DNA is isolated by the 16S rRNA gene area of 2 isolates with the universal primes 27F and 1492R and the sequence analysis was performed. With the Mega 7.0.18 package program, the maximum probability algorithm has been selected and filogenic trees have been created with the distance matrice of Jukes and Cantor. It has been found that all isolates are negative in the direction of the laccas enzyme, and the isolates N1 and N2 are positive in the direction of the mananase enzyme. In the philogenetic tree created for the N1 isolate, the Halomonas bacteria are accumulated with a strong homology among themselves. In the philogenetic tree created for N2 isolation, Bacillus bacteria are generally accumulated with a strong homology. The isolates of N1 and N2 were diagnosed as N1 Halomonas stevensii and N2 Bacillus agaradhaerens according to their morphological, physiological, biochemical properties and 16S rRNA gen area analysis.
The aim of this study is the characterization and diagnosis of bacteria that have the ability to produce laccase and mannanase enzymes from water and sediment samples collected from Lake Van. The material of this research were form water and sediment samples collected from Van Lake. The isolation study was carried out on the alkali medium by dilution plate method. Isolate with two different colony morphologies were observed. Colony diameter, pigmentation, cell shape, gram reaction, indole test, citrate test, voges-proskauer (vp), arabinose, mannitol, sucrose, adonitol, methyl_d-glycoside, cellobiosis, mannose, salicin, trahelosis, galactose, inositol, rafinose, inulin, sorbitol, xylose, melesitose, glucose, arginine, nitrate reduction, beta galactosidase, oxidase, laccase enzyme activity, mannanase enzyme activity, starch hydrolysis, tween 80 hydrolysis, skim milk hydrolysis and xylan hydrolysis tests have made. The ability of isolates to produce laccase enzyme was evaluated with 0.2% mM ABTS and 0.1% mM CuSO4. Their ability to produce mannanase enzyme was determined using an alkaline medium containing 1% mannose and phenol. 16S rRNA gene region of 2 isolates whose genomic DNA was isolated were amplified with universal primers of 27F and 1492R and sequence analysis was performed. Phylogenic trees were created with the distance matrix of Jukes and Cantor by selecting the Maximum Likelihood algorithm with the Mega 7.0.18 package program. It was observed that all of the isolates were negative in terms of laccase enzyme and N1 and N2 isolates were positive in terms of mannanase enzyme. In the phylogenetic tree created for N1 isolate, Halomonas bacteria was clustered with a strong homology among themselves. Bacillus bacteria were generally clustered with a strong homology in the phylogenetic tree created for N2 isolate. N1 and N2 isolates were diagnosed as N1 Halomonas stevensii and N2 Bacillus agaradhaerens according to their morphological, physiological, biochemical properties and 16S rRNA gene region analysis.
Alan : Fen Bilimleri ve Matematik; Mühendislik
Dergi Türü : Uluslararası
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