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

Abstract

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.

Keywords

• Recombinant interferon beta secretion,signal peptides,periplasmic protein

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

Abstract

: 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.

Keywords

• Rekombinant interferon beta salgılanması,sinyal peptidler,periplazmik protein

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