Characterization of recombinant soyacystatin expressed in E.coli

Rekombinant (r-) soyasitatinin papaine olan inhibitörü aktivitesi karakterize edilerek yumurta beyazı sistatin ile karşılaştmlmıştır. E.coli de sentez edilen r-soyasistatin 4 phenyl-Sepharose ve DEAE kolonları rekombinant protein olarak 4.33 kat saflaştırümıştır. İsaelektirik fokus jeli ile papain ve r-soyasistatin arasındaki spesifik interaksiyon tespit edilmiştir. Papain ve r-soyasistatinin bir kompleks oluşturduğu ve bu kompleksin pl si sistatin ve papain pl lar arasında çözündüğü tespit edilmiştir. Her iki sistatinde geniş bir pH aralığında (pH 4-10) stabilite göstermiş ve soyasistatinin termal stabiliteside yumurta beyazı sistatine benzer bulunmuştur test edilen sıcaklıklarda (0-100 °C). r-Soyasistatinin papaine karşı olan inhibitor aktivitesi yumurta beyazı sistatine benzer bulunmuştur.

Anahtar Kelimeler:

sistein proteinaz, inhibitör, proteinler, rekombinant protein, saflaştırma, papain, Escherichia coli

E.coli tarafından sentezlenen recombinant soyacystatinin karakterizasyonu

Recombinant (r-) soyacystatin was characterized for their inhibitory activity against papain and compared to egg white cystatin. r-Soyacystatin expressed in E. coli was purified 4.33 fold as a recombinant protein with phenyl-Sepharose and DEAE. Egg white cystatin was purified by using affinity chromatography on CM-papain-Sepharose. The specific interaction of r-soyacystatin and papain was detected on isoelectric focusing gel. Papain and r-soyacystatin formed a complex and the complex was resolved in between pis of cystatins and papain. Both cystatins showed high stability at the wide pH range (pH 4-10), and the thermal stability of soyacystatin was comparable at the temperature range (0-100 °C). The r-soyacystatin exhibited papain-like protease inhibition activity comparable to that of the egg white cystatin, which could inhibit papain. .

Keywords:

cysteine proteinases, inhibitors, proteins, recombinant proteins, purification, papain, Escherichia coli,

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Gaziosmanpaşa Üniversitesi Ziraat Fakültesi Dergisi-Cover

ISSN: 1300-2910
Yayın Aralığı: Yılda 3 Sayı
Başlangıç: 1985
Yayıncı: Tokat Gaziosmanpaşa Üniversitesi Ziraat Fakültesi Dergisi Yayın Ofisi