Fatma Matpan Bekler, Kemal Güven, Zindar Aslan

Bingöl Sıcak Su Kaplıcalarından İzole Edilen Bakterilerin Tanımlanması, Proteaz ve Amilaz Enzimlerinin Karakterizasyonu

Bingöl İli Binkap sıcak su kaynaklarından izole edilen bakteriler karakterize edilerek, bunlara ait endüstriyel alanda önem arz eden proteaz ve amilaz gibi enzimleri üzerinde çalışmalar yapılmıştır. Bu çalışmada su ve toprak örneklerinden iki bakteri suşu izole edildi. Bu bakterilerin çeşitli biyokimyasal testleri ile morfolojik ve fizyolojik analizleri yapıldı. İzole edilen 4NK bakterisinin çubuk şeklinde Gram pozitif olduğu, 5NK bakterisinin de Gram pozitif ve çubuk şeklinde olduğu belirlenmiştir. İzole edilmiş bu iki bakteri için optimum pH değerinin 6.0 olduğu tespit edildi. Optimum üreme sıcaklıkları ise sırasıyla 40 ve 45 °C olarak belirlenmiştir. İzole edilen bakterilerin optimum koşulları belirlendikten sonra bunların ürettikleri enzimlerin optimum koşulları belirlendi. Buna göre 4NK bakterisinin proteaz ve amilaz enzimlerini üretim açısından optimum süreleri sırasıyla 18. saatte (195.80 U mg-1) ve 15. saatte (428.33 U mg-1) olduğu belirlenmiştir. Proteaz ve amilaz enzimlerinin optimum pH’sı 8.0, optimum sıcaklıkları sırasıyla 40-50 °C ve 50-60 °C olarak bulunmuştur. Ayrıca, 5NK bakteri varyetesi için proteaz ve amilaz enzimlerini üretim açısından optimum süresi, proteaz için 260.93 U mg-1ve amilaz için 380.58 U mg-1olmak üzere 24. saat olarak belirlenmiştir. Proteaz ve amilaz enzimlerinin optimum pH’ları 8.0, optimum sıcaklıkları 50 °C olarak bulunmuştur. İzolasyonu yapılan 4NK ve 5NK bakterilerinin 16S rRNA dizi analizi yapılmış, buna göre 4NK bakterisinin Bacillus subtilis türüne yakın, 5NK bakterisinin ise Bacillus paralicheniformis türüne filogenetik olarak yakın olduğu tespit edilmiştir.

Anahtar Kelimeler:

Amilaz, proteaz, sıcak su kaynakları, enzim, Bacillus

Identification of Bacteria from Bingöl Hot Springs and Characterization of Protease and Amylase

Bacteria isolated from Binkap hot spring from Bingöl Province were characterised and studies were carried out on their industrially important enzymes such as protease and amylase. In this study, two bacteria were isolated in the water and soil samples. Various morphological, physiological and biochemical tests of these bacteria were applied. It was determined that the isolated 4NK bacteria were Gram positive in the form of rods, and the 5NK bacteria were Gram positive and rod shaped. It was determined that the optimum pH value for both isolated bacteria was 6.0. Optimum growth temperatures for 4NK and 5NK were determined as 40 °C and 45 °C, respectively. After determinaton of the optimum conditions for the isolated bacteria, the optimum conditions of the enzymes produced were also detected. Accordingly, the optimum production times of protease and amylase enzymes by 4NK bacteria were determined as 18th hour (195.80 U mg-1) and 15th hour (428. U mg-1), respectively. The optimum pH of protease and amylase enzymes were found to be 8.0, while optimum temperatures were as 40-50 °C and 50-60 °C, respectively. The optimum production time of protease and amylase enzymes by 5NK bacteria was determined to be 24 hours with activities of 260.93 U mg-1 for protease and 380.58 U mg-1for amylase. The optimum pH of protease and amylase enzymes was determined as 8.0 and their optimum temperature as 50 °C. 16S rRNA sequence analysis of isolated 4NK and 5NK bacteria was performed and it was determined 4NK bacteria were close to Bacillus subtilis and 5NK bacteria were phylogenetically close to Bacillus paralicheniformis.

Keywords:

Amylase, protease, enzyme, hot spring, Bacillus,

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