A Review on the Use of the Signal Peptide in Recombinant Interferon beta Secretion in Escherichia coli

Yıl 2019, Cilt: 45 Sayı: 1, 10 - 21, 30.04.2019

Mehmet Öztürk Yakup Ermurat

Öz

Eukaryotic and prokaryotic expression systems are used with similar
processes for the production of rIFN-β1a and rIFN-β1b which are from the
recombinant interferon beta (rIFN-β) protein varieties. The difficulties in
purification of the produced rIFN-β1a by eukaryotic expression systems and the
influence of its biological activity make recombinant protein production
difficult with these processes. In the production of rIFN-β1b, where
prokaryotic expression systems were used, purification and activity problems
were found to be less frequent. Better use of product secretion capability in Escherichia coli from prokaryotic
bacteria indicates that this system will be preferred in the production of
rIfN-β1b. If the construct is designed to secrete recombinant proteins out of
the host cell from which the recombinant proteins are produced, it is envisaged
that the host cells will not fragment during purification, thereby facilitating
product purification processes. The use of E.
coli with type I and type II secretion in the production of rIFN-β1b
proteins will reduce the purification and activity problems in the secretion of
rIFN-β1b produced in this host, as well as the toxic effect of rIFN-β1b to the
host cell is expected to diminish. To this end, various approaches are
contemplated to overcome the disadvantages of the E. coli production system, such as transferring the DmsA signal
peptide, the Tat-type signal peptide, to the expression vector pET22b, in
addition to the PelB signal peptide, the Sec signal peptide. In this review,
information about the properties of IFNs, the clinical applications of rIFN-β
and the researches based on the investigations of rIFN-β1b production  methods, and information about the use of
signal peptides in the process of producing rIFN-β1b in E. coli host cells were highlighted.

Anahtar Kelimeler

Recombinant interferon beta secretion, signal peptides, periplasmic protein

Escherichia coli’de Yenibileşenli İnterferon beta Salgılanmasında Sinyal Peptidi Kullanımı Üzerine bir Derleme

Öz

: Rekombinant
interferon beta (rİFN-β) proteini çeşitlerinden rİFN-β1a ve rİFN-β1b üretimi
için benzer süreçler ile ökaryotik ve prokaryotik ekspresyon sistemleri
kullanılmaktadır. Ökaryotik ekspresyon sistemleri ile üretilen rİFN-β1a’nın
saflaştırılmasında zorlukların yaşanması ve biyolojik aktivitesinin etkilenmesi
bu süreçler ile rekombinant protein üretimini güçleştirmektedir. Prokaryotik
ekspresyon sistemlerinin kullanıldığı rİFN-β1b üretiminde ise saflaştırma ve
aktivite sorunlarının daha az gerçekleştiği belirlenmiştir. Prokaryotik
bakterilerden Escherichia coli’de
bulunan ürün salgılama özelliğinin daha iyi kullanılması, bu sistemin rİFN-β1b
üretiminde daha fazla tercih edileceğini göstermektedir. Rekombinant
proteinlerin üretildikleri konakçı hücreden dışarı salgılatılacak şekilde yapı
tasarlanması durumunda, saflaştırma sırasında E. coli konakçı hücreler parçalanmayacağından ürün saflaştırma
işlemlerini kolaylaştıracağı öngörülmektedir. Tip I ve tip II salgılama
sistemine sahip olan E. coli’nin
rİFN-β1b proteinlerinin üretiminde kullanımı, bu konakçıda üretilen rİFN-β1b’nin
hücre dışına salgılatılmasında saflaştırma ve aktivite sorunlarını azaltacağı,
ayrıca rİFN-β1b’nin konakçı hücreye yapacağı toksik etkininde ortadan kalkacağı
tahmin edilmektedir. Bu amaçla E. coli
üretim sistemindeki dezavantajları gidermek için Sec sinyal peptidi olan PelB
sinyal peptidine ilave olarak Tat tipi sinyal peptidi olan DmsA sinyal
peptidi’nin pET22b ifade vektörüne aktarılmasıyla rİFN-β1b proteinlerinin hücre
dışına salgılatılması işlemi gibi çeşitli yaklaşımlar tasarlanmaktadır. Bu
derlemede, İFN’ların özelliklerine, rİFN-β’nın klinik uygulamalarına ve
rİFN-β1b üretim yöntemleri ile ilgili yapılmış araştırmalara dayalı bilgi ve ve
rİFN-β1b’nin E. coli konakçı
hücrelerinde üretilmesi sürecinde sinyal peptidlerinin kullanımı ile ilgili
bilgilere dikkat çekilmiştir.

Anahtar Kelimeler

Rekombinant interferon beta salgılanması, sinyal peptidler, periplazmik protein

Kaynakça

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