Optimization of Fermentation Parameters for Inulinase Production by Statistical Approach

Öz

The aim of the study was to carry out high-activity and low-cost inulinase production from Rhizopus oryzae HBF351, a new fungal strain, in solid phase fermentation. For this, it was firstly aimed to determine the best substrate that supports high enzyme production and to optimize some parameters that affect the fermentation condition by a statistical approach. Five different substrates (wheat bran, oat bran, onion peel, apple pulp and lettuce root powder) were investigated to find the best substrat supporting maximum enzyme productions. Box–Behnken design (BBD) was employed to optimize fermentation conditions (initial pH (4-7), incubation temperature (25°C-45°C), initial moisture level (50-100%) and incubation time(3-7 days)) for the production of inulinase. In the study, wheat bran was the best substrate that supports enzyme activity. pH, incubation temperature, moisture level and incubation time have been shown to be highly effective parameters on enzyme production (p <0.0001). In the study, the fitness of the model was evaluated using R2 (regression coefficient) (0.99) and a good correlation were obtained between experimental and predicted results. Optimum conditions providing maximum inulinase activity (385.1 IU gds-1) were found as pH 5.18, 33.4 °C, 72.1 % moisture level and 4.7 days. As a result, this study provides a cost-effective method to produce inulinase from Rhizopus oryzae. Also, it can be suggested that the produced inulinase has strong potential for usage in the production of fructose syrup and other industrial areas due to its high activity.

Anahtar Kelimeler

• Box-Behnken Design
• Inulinase
• Optimization
• Solid Phase Fermentation

İnülinaz Üretimi İçin Fermantasyon Parametrelerinin İstatistiksel Yaklaşımla Optimizasyonu

Öz

Çalışmanın amacı katı faz fermantasyonunda yeni bir fungus izolatı olan Rhizopus oryzae HBF351’dan aktivitesi yüksek ve düşük maliyetli inülinaz üretiminin gerçekleştirilmesidir. Bunun için ilk olarak yüksek enzim üretimini destekleyen en iyi substratın belirlenmesi ve fermantasyon ortamını etkileyen bazı parametrelerin istatistiksel bir yaklaşımla optimize edilmesi hedeflenmiştir. Maksimum enzim üretimini destekleyen en iyi ortamı bulmak için 5 farklı substrat (buğday kepeği, yulaf kepeği, soğan kabuğu, elma posası ve marul kökü tozu) araştırılmıştır. İnülinaz üretimi için bazı fermantasyon koşullarını (başlangıç pH (4-7), inkübasyon sıcaklığı (25°C-45°C), başlangıç nem seviyesi ( %50-100) ve inkübasyon süresi (3-7 gün)) optimize etmek amacıyla Box-Behnken tasarımı (BBD) kullanılmıştır. Çalışmada buğday kepeğinin enzim aktivitesini destekleyen en iyi substrat olduğu bulunmuştur. pH, inkübasyon sıcaklığı, nem seviyesi ve inkübasyon süresinin enzim üretimi üzerinde oldukça etkili parametreler olduğu gösterilmiştir (p<0.0001). Çalışmada modelin uygunluğu R2 (regresyon katsayısı) (0.99) kullanılarak değerlendirilmiştir ve deneysel ve tahmin edilen sonuçlar arasındaki iyi korelasyon elde edilmiştir. Maksimum inulinaz aktivitesini (385.1 IU gds-1) sağlayan optimum koşullar pH 5.18, 33.4 °C, % 72.1 nem seviyesi ve 4.7 gün olarak bulunmuştur. Sonuç olarak, bu çalışma Rhizopus oryzae'den inülinaz üretmek için uygun maliyetli bir yöntem sunmaktadır. Ayrıca, yüksek enzim aktivitesi elde edilen inülinazı, fruktoz şurubu ve diğer endüstriyel alanlarda kullanım için güçlü bir potansiyel aday yapmaktadır.

Anahtar Kelimeler

• İnülinaz
• Box-Behnken Tasarımı
• Katı Faz Fermantasyonu
• Optimizasyon

Kaynakça

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