Merve AYTEKİN AKBABA, Asliye KARAASLAN, Selin ALİHANOĞLU, Hasan VARDİN, Mehmet KARAASLAN

Farklı pH ve sıcaklık değerlerinin Pichia pastoris'de rekombinant ekspanzin üretimi üzerine etkisi

Bu çalışma kapsamında rekombinant Ekspanzin üretimi amacıyla domatesten izole edilen LeExp1 geni Pichia pastoris mayalarına pPIC9K vektörü vasıtasıyla aktarılmıştır. Ekspresyon koşulları 20, 25, 30°C ve pH 4.0, 5.0, 6.0 ve 7.0 değerleri için ayrı ayrı sağlanmıştır. Fermentasyon özütlerinin Biomass analizi ve Bradford assay deneyleri sonucunda toplam rekombinant protein miktarları ve toplam hücre konsantrasyonları belirlenmiştir. Farklı pH ve sıcaklık değerlerinin rekombinant protein üretim miktarına ve toplam hücre konsantrasyonuna olan etkileri incelenmiştir. Deneysel sonuçlara göre maksimum rekombinant ekspanzin üretimi için P. pastoris mayalarının optimum fermantasyon koşulları 25oC sıcaklık ve pH 7.0 olarak tespit edilmiştir.

Anahtar Kelimeler:

Ekspanzin, Pichia pastoris, Rekombinant protein, pH, sıcaklık

Effect of different pH and temperature on the production of recombinant expansin in Pichia pastoris

In this study LeExp1 gene isolated from tomato genome was transferred to Pichia pastoris by using pPIC9K vector for production of recombinant Expansin protein. Recombinant protein expression was carried out at 20, 25, 30°C temperature and 4.0, 5.0, 6.0 and 7.0 pH levels. Total protein and dry cell weight amounts was determined in fermentation medium. The effects of different fermentation temperature and pH level in the medium on protein expression and total cell concentration were investigated. The optimum fermentation conditions for P. pastoris were determined as 25oC and pH 7.0 according to experimental results.

Keywords:

Expansin, Pichia pastoris, Recombinant Protein, pH, Temperature,

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