Year 2017, Volume 38, Issue 4, Pages 86 - 97 2017-12-08

Optimization of Hnox Protein Production in Escherichia Coli
HNOK Proteinlerinin Escherichia Coli’de Üretiminin Optimizasyonu

Nur Başak SÜRMELİ [1]

183 367

Hemeproteins carry a variety of different functions in organisms ranging from steroid biosynthesis to respiration, signaling to drug metabolism. In industry, hemeproteins are used for production of drugs such as: pravastatin for lowering cholesterol, progesterone for hormonal treatment of cancers of uterus and cervix, and cortisone used against allergy and inflammation. Hemeproteins can also be used in drug development and biological remediation. The industrial applications of hemeproteins will expand with development of molecular biology and protein design techniques. One of the obstacles to the widespread use of hemeproteins is difficulties in production of high levels of heme bound protein. This study aims to maximize the amount of heme cofactor bound hemeprotein produced. Here, three important factors affecting hemeprotein production in bacteria are examined: induction by isopropyl β-D-1-thiogalactopyranoside (IPTG), δ-aminolevulinic acid (ALA), precursor for heme biosynthesis, and expression temperature. Effects of these factors on production of thermophilic hemeprotein TtHNOX are investigated. Since ALA is an expensive molecule, optimization of the amount of ALA used is important. The most suitable conditions to produce TtHNOX is at low temperature, 0.5 mM IPTG and 1 mM ALA. This study concludes that ALA concentration and expression temperature are important in production of heme bound hemeproteins.

Hemoproteinler canlılarda steroid biyosentezinden solunuma, sinyalizasyondan ilaç metabolizmasına kadar pek çok farklı biyolojik süreçlerde önemli görevler üstlenirler. Endüstride, hemoproteinler kolesterol düşürücü pravastatin, rahim ve rahim ağzı kanserlerinin hormonal tedavisinde kullanılan progesteron, alerji ve yangı’ya karşı kullanılan kortizon gibi ilaçların üretiminde kullanılmaktadır. Bunların yanı sıra hemoproteinlerin ilaç geliştirilmesi, biyolojik iyileştirme gibi alanlarda kullanılması da planlanmaktadır. Moleküler biyoloji ve protein tasarımı tekniklerinin gelişmesi ile bu proteinlerin endüstriyel uygulama alanları da genişleyecektir. Hemoproteinlerin bu alanlarda yaygın kullanımı karşısındaki en önemli engellerden biri Hem kofaktörüne bağlı bir şekilde yüksek miktarda hemoprotein üretilememesidir. Bu çalışmada Hem kofaktörüne bağlı bir şekilde üretilen hemoprotein miktarını en yüksek seviyeye getirebilecek koşullar araştırılmıştır. Bu çalışma kapsamında bakteride hemoprotein üretimini etkileyen en önemli üç etken olan indükleyici izopropil β-D-l-tiyogalaktopiranosid (IPTG), ve Hem öncül molekülü δ-aminolevülinik asit (ALA), ve ekspresyon sıcaklığı incelenmiştir. Bu etkenlerin termofilik hemoprotein TtHNOK üretimine etkisi araştırılmıştır. Özellikle, ALA pahalı bir molekül olduğu için hemoproteinlerin üretiminde kullanılan ALA miktarının optimizasyonu önemlidir. Bu çalışma sonucunda Escherichia coli bakterisinde Hem kofaktörüne bağlı TtHNOK proteinin üretimi için en uygun koşulların düşük sıcaklık, 0,5 mM IPTG ve 1 mM ALA olduğu gösterilmiştir. Bu çalışmada çıkan sonuçlar Hem kofaktörüne bağlı hemoprotein üretiminde ALA konsantrasyonun ve ekspresyon sıcaklığının önemli olduğunu göstermiştir. 

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Subjects Basic Sciences
Journal Section Natural Sciences
Authors

Author: Nur Başak SÜRMELİ (Primary Author)

Bibtex @research article { csj363308, journal = {Cumhuriyet Science Journal}, issn = {2587-2680}, eissn = {2587-246X}, address = {Cumhuriyet University}, year = {2017}, volume = {38}, pages = {86 - 97}, doi = {10.17776/csj.363308}, title = {Optimization of Hnox Protein Production in Escherichia Coli}, key = {cite}, author = {SÜRMELİ, Nur Başak} }
APA SÜRMELİ, N . (2017). Optimization of Hnox Protein Production in Escherichia Coli. Cumhuriyet Science Journal, 38 (4), 86-97. DOI: 10.17776/csj.363308
MLA SÜRMELİ, N . "Optimization of Hnox Protein Production in Escherichia Coli". Cumhuriyet Science Journal 38 (2017): 86-97 <http://dergipark.org.tr/csj/issue/32371/363308>
Chicago SÜRMELİ, N . "Optimization of Hnox Protein Production in Escherichia Coli". Cumhuriyet Science Journal 38 (2017): 86-97
RIS TY - JOUR T1 - Optimization of Hnox Protein Production in Escherichia Coli AU - Nur Başak SÜRMELİ Y1 - 2017 PY - 2017 N1 - doi: 10.17776/csj.363308 DO - 10.17776/csj.363308 T2 - Cumhuriyet Science Journal JF - Journal JO - JOR SP - 86 EP - 97 VL - 38 IS - 4 SN - 2587-2680-2587-246X M3 - doi: 10.17776/csj.363308 UR - https://doi.org/10.17776/csj.363308 Y2 - 2017 ER -
EndNote %0 Cumhuriyet Science Journal Optimization of Hnox Protein Production in Escherichia Coli %A Nur Başak SÜRMELİ %T Optimization of Hnox Protein Production in Escherichia Coli %D 2017 %J Cumhuriyet Science Journal %P 2587-2680-2587-246X %V 38 %N 4 %R doi: 10.17776/csj.363308 %U 10.17776/csj.363308
ISNAD SÜRMELİ, Nur Başak . "Optimization of Hnox Protein Production in Escherichia Coli". Cumhuriyet Science Journal 38 / 4 (December 2017): 86-97. https://doi.org/10.17776/csj.363308