Selülaz Enzimi Üreten Bakterilerin İzolasyonu ve Su Ürünleri Yetiştiriciliğinde Kullanılabilirliğinin Araştırılması

Bu çalışmada Çukurova Üniversitesi kampüsünün iki farklı lokasyonundan toplanan toprak örneklerinden karboksimetilselülaz (CMCaz) enzimini üreten bakterilerin izolasyonu gerçekleştirilmiştir. Bakteriler sırasıyla Bacillus sp. SU44 ve BK17 olarak isimlendirilmişlerdir. Bu mikroorganizmalardan hücredışı selülaz enziminin izolasyonu ve kısmi karakterizasyonu gerçekleşmiştir. Yapılan çalışmada her iki izolata ait enzimin optimum pH ve sıcaklık değerleri sırasıyla 5.0 ve 40 °C olarak belirlenmiştir. Bacillus sp. SU44 izolatı maksimum CMCaz üretim seviyesine inokülasyonun başlangıcından itibaren 24. saatte ulaşırken, BK17 izolatı 48. saatte ulaşmıştır. Termal kararlılık deneyleri her iki enzimin de 40 °C’den sonra kalan aktivite kaybına uğradığını ortaya koymuştur. CoCl her iki enzim üzerinde de inhibisyon etkisi gösterirken, EDTA, SDS, MgCl ve CaCl2 değişik oranlarda aktivatör olarak rol oynamışlardır. Her iki izolat da penisilin, ampisilin, gentamisin, tetrasiklin ve siprofloksasin antibiyotiklerine karşı değişik oranlarda hassasiyet göstermiştir. 16S rDNA dizileme analizi Bacillus sp. SU44 ve BK17 suşlarının sırası ile Bacillus pasificus ve Bacillus tropicus ile %99’luk benzerlik gösterdiği ortaya koymuştur.

Anahtar Kelimeler:

Bacillus pasificus, Bacillus tropicus, selülaz, izolasyon, karakterizasyon

Isolation of Cellulase Enzyme Producing Bacteria and Investigation of Usability in Aquaculture

In this study, bacteria producing carboxymethylcellulase (CMCase) enzyme were isolated from soil samples collected from two different locations of Çukurova University campus. Bacteria were named as Bacillus sp. SU44 and BK17, respectively. Isolation and partial characterization of the extracellular cellulase enzyme from these microorganisms were performed. In the study, the optimum pH and temperature values of the enzymes of both isolates were determined as 5.0 and 40 °C, respectively. Bacillus sp. SU44 isolate reached the maximum CMCase production level at 24 hours from the start of inoculation, while BK17 isolate reached 48 hours. Thermal stability experiments revealed that both enzymes lost residual activity after 40 °C. While CoCl showed an inhibitory effect on both enzymes, EDTA, SDS, MgCl2 and CaCl2 acted as activators at different rates. Both isolates showed varying sensitivity to penicillin, ampicillin, gentamicin, tetracycline and ciprofloxacin antibiotics. 16S rDNA sequencing analysis revealed that Bacillus sp. SU44 and BK17 strains showed 99% similarity with Bacillus pacificus and Bacillus tropicus, respectively.

Keywords:

Bacillus pasificus, Bacillus tropicus, cellulase, isolation, characterization,

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