Purification and determination of inhibitory activity of recombinant soyacystatin against papain and fish protease

Rekombinant (r-) soyasistatinin papaine olan inhibitor aktivitesi karakterize edilerek yumurta beyazı sistatini ile karşılaştırılmıştır. E.coli de sentezlenen r-soyasistatin 4.33 kez saf rekombinant protein biçiminde phenyl-Sepharose ve DEAE kolonları ile saflaştırılmıştır. Yumurta beyazı sistatin ise cm-papain-sepharose afînite kromatografısi kullanılarak saflaştırılm ıştır. Araştırmada, 2 ug papainin %50 sini inhibe etmek için gereken soyasistatin miktarı 0.245 ug ve yumurta beyazı sistatin miktarı ise 0.310 ug dır. r-Soyasistatın araştırmada kullanılan balık kasmdaki otolitik aktivitenin %90 nu inhibe etmiştir.

Rekombinant soyasistatinin saflaştırılması ve papaine ve balık proteazlarına karşı inhibitör aktivitesinin saptanması

Recombinant (r-) soyacystatin was characterized for its inhibitory activity against papain and compared to egg white cystatiri. r-Soyacystatin expressed in E. coli was purified with phenyl-Sepharose and DEAE 4.33 fold as a recombinant protein. Egg white cystatin was purified by using affinity chromatography on cm-papain-Sepharose. Inhibitory activity of r-soyacystatin was similar to that of egg white cystatin. The amount required to inhibit 50% activity of papain used in the assay, 2 ug, was 0.245 ug and 0.310 ug for soyacystatin and egg white cystatin, respectively. r-Soyacystatin inhibited 90% of autolytic activity in fish muscle.

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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