Optimization of Medium Components for Enhanced Expression of Recombinant Human Vascular Endothelial Growth Factor (VEGF165) in Kluyveromyces lactis GG799

Abstract

Vascular endothelial growth factor (VEGF165) serves as a vital biomarker, linked to diverse physiological processes and pathological conditions like cancer, owing to its pivotal role in angiogenesis. Therefore, an efficient method to obtain recombinant VEGF165 in high yield is required. This study focused on optimizing a medium to enhance the biomass of Kluyveromyces lactis (K. lactis) GG799 cells that express recombinant VEGF165. Through screening various media components, galactose, tryptone, and yeast extract were identified as the most effective components. Using the Box–Behnken design, the medium components were optimized, resulting in a notable 31% increase in biomass concentration. The optimized medium, comprising 45 g L-1 galactose, 30 g L-1 tryptone, and 5 g L-1yeast extract, achieved a biomass productivity (P) of 0.26 gDCW L-1 h-1. Additionally, the expression of VEGF165 reached 1024.09 μg per L of the culture medium, representing a 1.27-fold increase compared to the unoptimized medium. These results imply that the proposed optimized medium can serve as a model for a process development strategy, providing a foundation for obtaining elevated yields of other recombinant proteins in K. lactis.

Keywords

• Vascular endothelial growth factor (VEGF165)
• Kluyveromyces lactis
• Biomass optimization
• Recombinant protein
• Response Surface Methodology

Optimization of Medium Components for Enhanced Expression of Recombinant Human Vascular Endothelial Growth Factor (VEGF165) in Kluyveromyces lactis GG799

Öz

Vasküler endotelyal büyüme faktörü (VEGF165), anjiyogenezdeki rolünden dolayı çeşitli fizyolojik süreçler ve kanser gibi patolojik hastalıklar ile ilişkili önemli bir biyobelirteçtir. Bu nedenle, yüksek verimde rekombinant VEGF165 elde etmek için etkili bir yöntem gereklidir. Bu çalışmada, Kluyveromyces lactis (K. lactis) GG799 hücrelerinde rekombinant VEGF165 ekspresyonunu artırmak amacıyla büyüme ortamının optimize edilmesi amaçlanmıştır. Farklı ortam bileşenlerinin taranması sonucunda galaktoz, tripton ve maya ekstraktı en etkili bileşenler olarak belirlenmiştir. Box-Behnken tasarımı kullanılarak seçilen ortam bileşenleri optimize edilmiş ve biyokütle konsantrasyonunda %31'lik kayda değer bir artış sağlanmıştır. 45 g L-1 galaktoz, 30 g L-1 tripton ve 5 g L-1 maya ekstraktı içeren optimize ortam ile 0,26 gDCW L-1 h-1 biyokütle üretkenliğine (P) ulaşılmıştır. Ayrıca VEGF165 ekspresyonu, optimize edilmemiş ortama kıyasla 1,27 kat artışla 1024,09 μg L-1 değerine ulaşmıştır. Elde edilen sonuçlar, önerilen optimize edilmiş ortamın K. lactis'te diğer rekombinant proteinlerin üretimine yönelik proses geliştirme stratejisi olarak bir model olabileceğini göstermektedir.

Anahtar Kelimeler

• Vasküler endotelyal büyüme faktörü (VEGF165)
• Kluyveromyces lactis
• Biyokütle optimizasyonu
• Rekombinant protein
• Yanıt Yüzey Metodu

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