Tepki Yüzey Metodu ile Aeromonas caviae LipT51'den Ekstrasellüler, Termo-Alkali Lipaz Üretiminin İstatistiksel Optimizasyonu

Öz

Aeromonas caviae LipT51'den ekstrasellüler termo-alkali lipaz üretimi, tepki yüzey metodu (response surface methodology: RSM) ile istatistiksel olarak optimize edilmiştir. İlk olarak, en yüksek lipaz üretimi için farklı karbon (zeytinyağı, tributirin, ayçiçek yağı, atık kızartma yağı, gliserol, Tween 80, Tween 20, palmiye yağı ve Triton X100) ve azot (pepton, maya özütü, tripton, peynir altı suyu, üre, NaNO2, NH4NO3) kaynaklarını taramak üzere tek seferde bir faktör yöntemi uygulandı. Ardından, karbon kaynağı olarak seçilen atık kızartma yağı, azot kaynağı olarak seçilen tripton ve başlangıç pH’sı için optimum değerler Box-Behnken tasarımı (BBD) kullanılarak RSM ile belirlenmiştir. Lipaz üretimi için BBD'nin ikinci dereceden modeli istatistiksel olarak anlamlı ve güvenilir bulundu (p <0.0001, R2 = 0.9881). Maksimum lipaz üretimi (1.6 U mL-1) için doğrulanmış optimum koşullar, % 1.13 atık kızartma yağı, % 1.5 tripton ve pH 7.9 olarak belirlenmiştir. İlk kez bu çalışmada, bir A. caviae suşundan lipaz üretiminin optimizasyonu, ucuz atık malzeme kullanılarak optimize edilmiş kültür koşulları altında gerçekleştirildi. Deterjan aktivitesi ile değerli olduğu bilinen lipaz enziminin üretim verimliliği, optimize edilmeyen koşullara göre 2.7 kat arttı.

Anahtar Kelimeler

• Aeromonas caviae
• lipaz
• atık kızartma yağı
• optimizasyon
• tepki yüzey metodu
• BoxBehnken tasarımı

Statistical Optimization of Extracellular Thermo-Alkaline Lipase Production from Aeromonas caviae LipT51 with Response Surface Methodology

Öz

Extracellular thermo-alkaline lipase production from Aeromonas caviae LipT51 was statistically optimized by response surface methodology (RSM). First, the one factor at a time approach was implemented to screen the sources of carbon (olive oil, tributyrin, sunflower oil, waste frying oil, glycerol, Tween 80, Tween 20, palm oil, and Triton X100) and nitrogen (peptone, yeast extract, tryptone, whey, urea, NaNO2, NH4NO3) for the highest lipase production. Then, optimum values for waste frying oil selected as carbon source, tryptone selected as nitrogen source and initial pH of the medium were determined by RSM using Box-Behnken design (BBD). The quadratic model of BBD for lipase production was statistically significant and reliable (p < 0.0001, R2 = 0.9881). The validated optimal conditions for maximum lipase production (1.6 U mL-1) were determined as 1.13% waste frying oil, 1.5% tryptone and pH 7.9. For the first time in this study, optimization of lipase production from an A. caviae strain was carried out and under optimized culture conditions using cheap waste material. The production efficiency of lipase enzyme, which is known to be valuable with its detergent activity, increased 2.7 times compared to non-optimized conditions.

Anahtar Kelimeler

• Aeromonas caviae
• lipase
• optimization
• response surface methodology
• Box-Behnken Design

Kaynakça

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