Yeni izole edilmiş trametes trogii'nin lakkaz üretim kapasitesinin araştırılması: lakkaz enziminin tutuklanması ve karakterizasyonu.
Küçük Resim Yok
Tarih
2025
Yazarlar
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Dergi ISSN
Cilt Başlığı
Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Yeni izole edilmiş Trametes trogii suşundan lakkaz üretmek için farklı fermentasyon yöntemleri kullanıldı. Bu suş özellikle indükleyici olarak bakır içeren besiyerlerinde yüksek miktarda lakkaz üretmiştir. Katı faz fermentasyon sürecinde bakır içermeyen besiyerlerinde bile yüksek düzeyde lakkaz üretebilmiştir. Tüm ortamlarda tek bir lakkaz bandı gözlemlenmiştir. Çalışmada öncelikle, katı faz fermentasyonu sürecinden elde edilen ham serbest lakkaz enziminin, optimum sıcaklık ve pH değerleri saptanmıştır. Daha sonra, ham lakkaz enzimi bakteriyel selüloza tutuklanmış ve tutuklanmış lakkazın optimum sıcaklık ve pH değerleri saptanmıştır. Serbest lakkaz aktivitesinin 25 °C'den itibaren geniş bir sıcaklık aralığında yüksek olduğu gözlemlenirken, optimum pH değerinin de 3 olduğu belirlenmiştir. Tutuklanmış lakkazın optimum sıcaklığı ise, 40 °C ve pH değeri de 4 olarak saptanmıştır. Hem serbest hem de tutuklanmış lakkazın sıcaklık, pH, organik çözücüler, ağır metaller ve sodyum klorüre karşı kararlılığı ve ayrıca depolama kararlılığı saptanmıştır. İnhibitörlerin serbest ve tutuklanmış lakkaz aktivitesine etkisi de araştırılmıştır. Tutuklanmış enzim iyi bir operasyonel kararlılık göstermiştir. Çalışmada serbest ve tutuklanmış lakkazın boya renk giderim aktivitesi de araştırılmıştır. Serbest lakkaz Reaktif Mavi 171, Reaktif Mavi 198 ve Reaktif Mavi 19'un rengini farklı oranlarda giderebilmiştir. Tutuklanmış lakkazın tekrarlı kullanımı sırasında Reaktif Mavi 171 ve Reaktif Mavi 198'in rengi yüksek oranda giderilebilmiş ve tutuklanmış lakkaz miktarına bağlı olarak tekrarlı kullanım sayısı değişmiştir. Reaktif Mavi 19 için ise tekrarlı kullanım etkinliğinin düşük olduğu belirlenmiştir. Sonuç olarak; bu suş ile yüksek düzeyde lakkaz enzimi üretilebilir. Serbest lakkaz, biyoremediasyon uygulamalarında kullanılabilir. Ham lakkaz enzimi de doğal bir polimer olan bakteriyel selüloza tutuklanarak, kararlı bir tutuklanmış lakkaz elde edilebilir ve çeşitli kirleticilerin gideriminde kullanılabilir. Anahtar Kelimeler: Bakteriyel selüloz, enzim karakterizasyonu, fermentasyon, lakkaz, Trametes trogii, tutuklama.
Different fermentation methods were used to produce laccase by newly isolated Trametes trogii strain. This strain produced high amounts of laccase especially in copper-containing media, as an inducer. It was able to produce high levels of laccase even in copper-free media during the solid state fermentation process. A single laccase band was observed in all media. In the study, firstly, the optimum temperature and pH values of the crude free laccase enzyme obtained from the solid state fermentation process were determined. Then, the crude laccase enzyme was immobilized on bacterial cellulose (BC) and optimum temperature and pH values for immobilized laccase were determined. While the free laccase activity was observed to be high in a wide temperature range starting from 25 °C, the optimum pH value was determined to be 3. The optimum temperature for immobilized laccase was determined as 40 °C and the pH value as 4. The stability of both free and immobilized laccase against temperature, pH, organic solvents, heavy metals and sodium chloride as well as storage stability were determined. The effect of inhibitors on free and immobilized laccase activity was also investigated. The immobilized enzyme showed good operational stability. The dye decolorization activity of free and immobilized laccase was also investigated. Free laccase was able to remove the color of Reactive Blue 171, Reactive Blue 198 and Reactive Blue 19 to different extents. The color of Reactive Blue 171 and Reactive Blue 198 could be removed to a high extent during repeatedly use of the immobilized laccase and the number of reuse varied depending on the amount of immobilized laccase used. It was determined that the repeated use activity was low for Reactive Blue 19. As a result; high levels of laccase enzyme could be produced with this strain. Free laccase can be utilized in bioremediation applications. The raw enzyme could be immobilized on BC, a natural polymer, and a stable immobilized laccase can be obtained and used in the removal of various pollutants. Keywords: Bacterial cellulose, enzyme characterization, fermentation, laccase, Trametes trogii, immobilization.
Different fermentation methods were used to produce laccase by newly isolated Trametes trogii strain. This strain produced high amounts of laccase especially in copper-containing media, as an inducer. It was able to produce high levels of laccase even in copper-free media during the solid state fermentation process. A single laccase band was observed in all media. In the study, firstly, the optimum temperature and pH values of the crude free laccase enzyme obtained from the solid state fermentation process were determined. Then, the crude laccase enzyme was immobilized on bacterial cellulose (BC) and optimum temperature and pH values for immobilized laccase were determined. While the free laccase activity was observed to be high in a wide temperature range starting from 25 °C, the optimum pH value was determined to be 3. The optimum temperature for immobilized laccase was determined as 40 °C and the pH value as 4. The stability of both free and immobilized laccase against temperature, pH, organic solvents, heavy metals and sodium chloride as well as storage stability were determined. The effect of inhibitors on free and immobilized laccase activity was also investigated. The immobilized enzyme showed good operational stability. The dye decolorization activity of free and immobilized laccase was also investigated. Free laccase was able to remove the color of Reactive Blue 171, Reactive Blue 198 and Reactive Blue 19 to different extents. The color of Reactive Blue 171 and Reactive Blue 198 could be removed to a high extent during repeatedly use of the immobilized laccase and the number of reuse varied depending on the amount of immobilized laccase used. It was determined that the repeated use activity was low for Reactive Blue 19. As a result; high levels of laccase enzyme could be produced with this strain. Free laccase can be utilized in bioremediation applications. The raw enzyme could be immobilized on BC, a natural polymer, and a stable immobilized laccase can be obtained and used in the removal of various pollutants. Keywords: Bacterial cellulose, enzyme characterization, fermentation, laccase, Trametes trogii, immobilization.
Açıklama
Anahtar Kelimeler
Biyoloji, Biology, Biyoteknoloji
