GFP Raportör Proteini Kullanarak E. coli'de Rekombinant Protein Üretiminde Peynir Altı Suyu Tozunun İndükleyici Rolünün Doğrulanması

Abstract

Peynir altı suyu, süt endüstrisinden elde edilen değerli bir yan üründür. Son zamanlarda yapılan çalışmalar, peynir altı suyunun rekombinant protein üretiminde hem indükleyici hem de karbon kaynağı olarak kullanımına odaklanmaktadır. Bu çalışmanın amacı, Escherichia coli'den üretilen rekombinant yeşil floresan proteininin indükleyicisi olarak tatlı peynir altı suyu tozunda bulunan laktozun etkinliğini ve minimum konsantrasyonunu değerlendirmektir. Peynir altı suyunun potansiyeli optik yoğunluk, hücre biyokütlesi, elektroforez analizi, floresans, protein saflığı temelinde değerlendirilmiş ve sonuçlar IPTG'den elde edilenlerle karşılaştırılmıştır. En yüksek floresans değerleri 20 mM laktoz eşdeğerinde deproteinize peynir altı suyu tozu ile elde edilmiştir. 2 mM laktoz içeren deproteinize peynir altı suyu tozu kullanımı, 0,5 mM IPTG'ye benzer miktarlarda rekombinant GFP üretmek için yeterli olmuştur. Peynir altı suyu tozunun bir indükleyici olarak doğrulanması, yüksek miktarda IPTG kullanımına bağlı toksisite ve ekonomik sorunları ortadan kaldıracaktır. Ayrıca, bu strateji peynir altı suyunun neden olduğu çevre kirliliği sorunları için alternatif bir çözüm sunmaktadır.

Keywords

• peynir altı suyu
• rekombinant protein üretimi
• yeşil floresan protein

Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein

Abstract

Whey is a valuable by-product obtained from the dairy industry. Recently studies focus on utilization of whey as both inducer and carbon source in recombinant protein production. The aim of the present study was to evaluate the efficiency and minimum concentration of lactose present in sweet whey powder as inducer of recombinant green fluorescent protein produced from Escherichia coli. The potential of whey was evaluated based on optical density, cell biomass, electrophoresis analysis, fluorescence, protein purity and results were compared with those obtained from IPTG. The highest fluorescence values were obtained with 20 mM lactose equivalent of deproteinized whey powder. Use of 2 mM lactose containing deproteinized whey powder was sufficient to produce recombinant GFP at comparable amounts to 0.5 mM IPTG. Validation of whey powder as an inducer will eliminate the toxicity and economic problems associated with high amount of IPTG usage. Furthermore, this strategy provides an alternative solution for the environmental pollution problems caused by whey.

Keywords

• sweet whey
• green fluorescent protein
• valorization
• recombinant protein expression

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