MEDIUM OPTIMIZATION FOR PROTEASE PRODUCTION VIA MODERATELY HALOTOLERANT VIRGIBACILLUS PANTOTHENTICUS USING RESPONSE SURFACE METHODOLOGY

Year 2020, , 69 - 79, 31.01.2020

Gizem Banger , Emrah Nikerel

https://doi.org/10.18036/estubtdc.680602

Abstract

Virgibacillus pantothenticus is an industrially promising, yet scarcely studied, moderately halotolerant microorganism (up to 10% NaCl) with high activity protease production potential. Following Response Surface Methodology, we employed a Central Composite Design for the experiments and constructed a second order polynomial model to represent the resulting data. For medium optimization for protease production we optimized the resulting model. 32 experiments (following the central composite design scheme) where, carbon (glucose), nitrogen (ammonium sulfate), potassium (potassium phosphate monobasic) and magnesium (magnesium sulfate) sources were studied in the media. Bacterial growth, residual glucose and protease activities were determined at the 48th hour of each experiment. The model was optimized and the concentrations were found for each parameter. Under the optimum conditions, the predicted protease activity is also experimentally verified and the model prediction was in very good agreement with experimental results. The interactions between medium components and their effect on cell growth and protease production are also sought. This work reports the improvements on protease production of a potentially interesting industrial host, Virgibacillus pantothenticus.

Keywords

Virgibacillus pantothenticus, Response Surface Methodology, Alkali Protease, Medium optimization

ILIMLI HALOFILIK VIRGIBACILLUS PANTOTHENTICUS’TA PROTEAZ ÜRETIMI IÇIN, YANIT YÜZEY YÖNTEMI ILE BESIYERI OPTIMIZASYONU

Abstract

Virgibacillus pantothenticus endüstriyel olarak umut vadeden, halotolerant (%10’a kadar tuzu tolere edebilen), yüksek aktiviteli proteaz üreticisi olma potansiyeli olan bir organizmadır. Bu çalışmada, Yanıt Yüzey Yöntemi metodu takip edilerek, deney tasarımı için Merkezi Kompozit Tasarımı, model olarak da ikinci derece polinom kullanılarak, biyokütle ve proteaz üretimi amaç fonskiyonu ile besiyeri optimizasyonu gerçekleştirilmiştir. Yanıt yüzeyi modelinin parametrelerinin belirlenmesi için, 32 deneyde faklı karbon (glukoz), azot (amonyum sülfat), potasyum (potasyum fosfat monobazik) ve magnezyum (magnezyum sülfat) miktarları, besiyerlerinin hazırlanması için kullanılmıştır. Her bir deneyde, bakterilerin büyümesi, 48 saat sonunda kalan glukoz miktarı ve proteaz aktivitesi belirlenmiştir. Kurulan model optimize edilmiş ve her bir besiyeri bileşeninin optimum seviyesi belirlenmiştir. Bu koşullarda, model tahmini deneysel olarak çok yakın bir sonuçla teyit edilmiştir. Aynı zamanda, besiyeri bileşenlerinin karşılıklı etkileşimleri ve hücre büyümesi ile proteaz üretimi üzerindeki etkileri araştırılmıştır. Bu çalışma, proteaz üreticisi olarak Virgibacillus pantothenticus’un potansiyelini geliştirmiştir.

Keywords

: Virgibacillus pantothenticus, besiyeri optimizasyonu, Yanıt Yüzey Yöntemi, Alkalen Proteaz

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