Yeni İzole Edilen Trametes versicolor'un Kesikli, Tekrarlı-Kesikli ve Katı-Faz Fermentasyon Süreçlerinde Lakkaz Üretimi

Abstract

Bu çalışmada yeni izole edilmiş Trametes versicolor suşunun lakkaz üretim yeteneği üç farklı fermentasyon sürecinde araştırılmıştır. Üç fermentasyon sürecinde de fungus lakkaz enzimini üretebilmiştir. Katı-faz fermentasyonu sürecinde 10 mM bakır içeren ortamda üretimin 20. gününde 13.21 U/mL lakkaz aktivitesi izlenmiştir, bu değer 0.5 mM ABTS+10 mM bakır içeren ortamda ise 27.30 U/mL’ye ulaşmıştır. Sıvı kesikli fermentasyon sürecinde ise 1 mM bakır içeren ortamda bakır içermeyen ortama göre lakkaz aktivitesi önemli oranda indüklenmiş ve lakkaz aktiviteleri 3, 6 ve 9. günlerde sırasıyla 2.25, 19.83 ve 24.57 U/mL’ye ulaşmıştır. ABTS ve ksilidin bu suşun lakkaz üretimini bakıra göre çok daha düşük düzeyde indüklemiştir. Sıvı tekrarlı-kesikli süreç de lakkaz üretimini önemli oranda indüklemiştir. Bakır içermeyen ortamda düşük düzeyde enzim aktiviteleri saptanırken, bakır içeren ortamda lakkaz aktiviteleri indüklenmiş ve peletlerin 6. kullanımında aktivite 0.66 U/mL’den 9.87 U/mL’ye ulaşmıştır. Tüm fermentasyon süreçlerinde, bakırın lakkaz üretimi için önemli bir indükleyici olduğu gözlenmiştir ve doğal poliakrilamid jel elektroforezi sonrası yapılan aktivite boyamaları aktif lakkaz bantlarını net olarak göstermiştir. Sonuçlar, bu suşun iyi bir lakkaz üreticisi olduğunu ve fermentasyon sürecine, üretim zamanına ve kullanılan indükleyiciye bağlı olarak lakkaz üretim veriminin değiştiğini ortaya koymuştur.

Keywords

• Bakır
• Enzim
• İnkübasyon
• İndükleyici
• Beyaz çürükçül fungus
• Zimogram

Laccase Production of Newly Isolated Trametes versicolor under Batch, Repeated-Batch, and Solid-State Fermentation Processes

Abstract

In this study, the laccase production ability of the newly isolated Trametes versicolor strain was investigated in three different fermentation processes. In all three fermentation processes, the fungus was able to produce the laccase enzyme. During the solid-state fermentation process 13.21 U/mL laccase activity was detected on the 20th day in the 10 mM copper-containing medium, while this value reached to 27.30 U/mL in the medium containing 0.5 mM ABTS+10 mM copper. During the liquid batch fermentation process, laccase activity was significantly induced in the medium containing 1 mM copper and the laccase activities reached 2.25, 19.83 and 24.57 U/mL compared to the medium without copper on the 3rd, 6th, and 9th days, respectively. ABTS and xylidine induced the laccase production of this strain at a much lower level than copper. The liquid repeated-batch process also significantly induced the laccase production. While low level of enzyme activities were detected in a copper-free medium, laccase activities were induced in the copper-containing medium and the activity increased from 0.66 U/mL to 9.87 U/mL at the 6th use of the pellets. Copper was detected as an effective inducer for laccase production in all fermentation processes and activity staining after native polyacrylamide gel electrophoresis clearly showed the active laccase bands. The results revealed that this strain is a good laccase producer and the laccase production yield varies depending on the fermentation process, production time, and inducer used.

Keywords

• Copper
• Enzyme
• Incubation
• Inducer
• White rot fungus
• Zymogram

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