Optimization of Thermotolerant Cellulase from Streptomyces sp. M127-1 with Response Surface Methodology using Solid-State Fermentation

Yıl 2024, Cilt: 24 Sayı: 1, 217 - 227, 27.02.2024

Muzaffer Binek Elif Esin Hameş

https://doi.org/10.35414/akufemubid.1336051

Öz

The study aims to optimise cellulase that can be used in composting from actinomycete isolate using the solid-state fermentation method with Response Surface Methodology and its partial characterisation. Five isolates (M127-1, M127-2B, 1M1, M45-1 and M6c-1) were previously determined to be candidate cellulase producers, were investigated for cellulase production by solid-state fermentation, and isolate M127-1 was selected for further studies. For solid substrate selection, solid-state fermentation was performed by isolate M127-1 in media-prepared combinations of different solid substrates (sawdust, soy flour, malt grass and wheat bran), and the highest enzyme activity was found to be wheat bran-malt grass (BM) medium (0.704 U/ml). Then, moisture, inoculum size, and incubation time were optimised with the central composite design for cellulase optimisation using BM medium. Optimum cellulase production conditions were determined as 79.93% moisture, 8.58 days incubation time and 8.38 (v/w) inoculum size, and it was observed that the enzyme activity increased by 1.8 times. The optimum pH of the enzyme was 6, and its temperature was 60C. The isolate was identified as Streptomyces sp. M127-1 by 16SrDNA sequence analysis. In conclusion, the thermotolerant cellulase production at high temperatures was optimised from Streptomyces sp. M127-1 through solid-state fermentation using cost-effective substrates.

Anahtar Kelimeler

Cellulose, Streptomyces, Composting, Solid state fermentation, Response surface methodology

Proje Numarası

FYL-2020-22356

Katı Hal Fermantasyonu Kullanılarak Streptomyces sp. M127-1’den Elde Edilen Termotolerant Selülazın Yanıt Yüzeyi Metodolojisi ile Optimizasyonu

Öz

Çalışma, kompostlamada kullanılabilecek selülazın aktinomiset izolatından katı hal fermantasyon yöntemiyle Yanıt Yüzeyi Metodolojisi (RSM) kullanılarak optimizasyonunu ve enzimin kısmi karakterizasyonunu amaçlamaktadır. Selülaz üreticisi olduğu belirlenmiş 5 izolat (M127-1, M127-2B, 1M1, M45-1 ve M6c-1) katı hal fermantasyonu ile selülaz üretimi açısından değerlendirilmiş ve izolat M127-1 ileri çalışmalar için seçilmiştir. Katı substrat seçimi için farklı substratların (talaş, malt çimi, soya unu ve buğday kepeği) kombinasyonları ile hazırlanan ortamlarda izolat M127-1 ile katı hal fermantasyonu gerçekleştirilmiş ve en iyi enzim aktivitesinin buğday kepeği-malt çimi (BM) (0,704 U/ml) ortamında olduğu belirlenmiştir. Ardından BM ortamı kullanılarak selülaz üretim optimizasyonu için nem, aşı miktarı ve inkübasyon süresi merkezi kompozit tasarımı kullanılarak optimize edilmiştir. Optimum selülaz üretim koşulları % 79,93 nem, 8,58 gün inkübasyon süresi ve 8,38 (v/w) aşı miktarı olarak belirlenmiş ve enzim aktivitesinin 1,8 kat arttığı görülmüştür. Enzimin optimum pH’sı 6 ve optimum sıcaklığı 60C olarak belirlenmiştir. 16SrDNA dizi analizi ile izolat, Streptomyces sp. M127-1 olarak tanımlanmıştır. Sonuç olarak yüksek sıcaklıkta aktif selülazın uygun maliyetli substratlar kullanılarak katı hal fermantasyonu ile Streptomyces sp. M127-1’den istatistiksel üretim optimizasyonu gerçekleştirilmiştir.

Anahtar Kelimeler

Selülaz, Streptomyces, Kompostlama, Katı hal fermentasyonu, Yanıt yüzey metodolojisi

Destekleyen Kurum

Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü

Proje Numarası

FYL-2020-22356

Teşekkür

Bu çalışma Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü FYL-2020-22356 nolu proje ile desteklenmiştir. Yazarlar çalışmaya olan desteklerinden dolayı Prof. Dr. Sait SARGIN’a teşekkür ederler.

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