Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi

ÇALIK, Pınar

Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi

Yürütücü

Pınar ÇALIK (Orta Doğu Teknik Üniversitesi, Mühendislik Fakültesi, Kimya Mühendisliği Bölümü, Ankara, Türkiye)

9 4

Proje Grubu: KBAG Sayfa Sayısı: 143 Proje No: 114R091 Proje Bitiş Tarihi: 01.05.2018 Metin Dili: Türkçe İndeks Tarihi: 16-03-2020

Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi

Öz:

Insan büyüme hormonu (hGH), 22 kDa molekül agırlıgında, 191 amino asitten olusan, beynin ön lobundaki hipofiz bezi tarafından üretilen bir proteindir. Hipofiz bezlerindeki somatotropik hücreler tarafından dogal olarak sentezlenen, kanda bulunan hGH?nin, insan organizmasında eksik, bozuk (defective) üretiminin neden oldugu problemlerin düzeltilmesi için dısarıdan verilme ihtiyacı ?diger hormonlar gibi- bu farmasötik temel maddenin biyoteknolojik prosesle endüstriyel üretimini önemli yapmaktadır. Bu proje kapsamında, gliseraldehit-3-fosfat dehidrojenaz baslatıcısının (promotör) (PGAP) üzerinde bulunan transkripsiyon faktörlerinin baglanma bölgelerini transkripsiyon mühendisligi ile modifiye ederek rekombinant protein (rprotein) üretimini artırmak hedeflenmistir. Pichia pastoris?te en çok kullanılan baslatıcılardan biri olan PGAP, olası transkripsiyon faktörlerinin (TF) baglanma bölgeleri açısından analiz edilmistir. Bulunan TF baglanma bölgeleri silinerek ya da dogal konumlarında duplike edilerek PGAP varyant kütüphanesi olusturulmustur. Aynı zamanda, seçilen TF?ler asırı-ekspres edilerek ya da silinerek, bu faktörlerin r-protein üretimine etkisi arastırılmıstır. Gelistirilen PGAP varyantları altında modifiye yesil floresan proteini üretiminin ekspresyon seviyeleri, dogal PGAP?a kıyasla 0.35-3.10 kat arasında degisim göstermistir. En yüksek ekspresyon seviyelerine ulasan baslatıcı varyantları olan Pro9 (Gal4-benzeri TF baglanma bölgesinin duplike edildigi) ve Pro10 (Rfx1 baglanma bölgesinin silindigi) kontrolü altında ikinci model protein, rekombinant insan büyüme hormonu (rhGH), ekspres edilmis; dogal PGAP?a kıyasla, sırasıyla, 2.4 ila 1.6 kat daha fazla rhGH üretilmistir. Yarı-kesikli biyoreaktör isletim kosullarında, Pro9 kontrolü altında ve Gal4-benzeri TF?nin asırı-ekspres edildigi sus ile rhGH üretiminde birim hücre basına 2.2 kat artıs elde edilmistir. Gelistirilen hibrit yarı-kesikli isletim ile en yüksek rhGH derisimi t = 8 st?te, CrhGH = 611 mg dm-3 elde edilmistir.

Anahtar Kelime: biyoreaktör isletim kosulları ortam tasarımı promotör Pichia pastoris Insan büyüme hormonu

Konular: Kimya, Uygulamalı Kimya, Tıbbi Biyoteknoloji ve Uygulamalı Mikrobiyoloji

Erişim Türü: Erişime Açık

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APA	ÇALIK P (2018). Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi. , 1 - 143.
Chicago	ÇALIK Pınar Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi. (2018): 1 - 143.
MLA	ÇALIK Pınar Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi. , 2018, ss.1 - 143.
AMA	ÇALIK P Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi. . 2018; 1 - 143.
Vancouver	ÇALIK P Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi. . 2018; 1 - 143.
IEEE	ÇALIK P "Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi." , ss.1 - 143, 2018.
ISNAD	ÇALIK, Pınar. "Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi". (2018), 1-143.

APA	ÇALIK P (2018). Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi. , 1 - 143.
Chicago	ÇALIK Pınar Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi. (2018): 1 - 143.
MLA	ÇALIK Pınar Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi. , 2018, ss.1 - 143.
AMA	ÇALIK P Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi. . 2018; 1 - 143.
Vancouver	ÇALIK P Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi. . 2018; 1 - 143.
IEEE	ÇALIK P "Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi." , ss.1 - 143, 2018.
ISNAD	ÇALIK, Pınar. "Metabolik Mühendislik ve Reaksiyon Mühendisigi Prensipleriyle Insan Büyüme Hormonu Üretimi Için Yeni Biyoteknolojik Proses Gelistirilmesi". (2018), 1-143.