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

Year 2018, Volume: 22 Issue: 2, 207 - 214, 25.06.2018

Merve Aytekin Akbaba Asliye Karaaslan Selin Alihanoğlu Hasan Vardin , Mehmet Karaaslan

https://doi.org/10.29050/harranziraat.315532

Abstract

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 25^oC sıcaklık ve pH 7.0 olarak tespit
edilmiştir.

Keywords

Ekspanzin, Pichia pastoris, Rekombinant protein, pH, sıcaklık

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

Abstract

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 25^oC
and pH 7.0 according to experimental results. 

Keywords

Expansin, Pichia pastoris, Recombinant Protein, pH, Temperature

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