SavedAdd your journal

Journals / Gaziosmanpaşa Üniversitesi Ziraat Fakültesi Dergisi / 2004 / Cilt: 21 - Sayı: 1

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

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

Özlem AKPINAR
Pages
19–26
DOI
—
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

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.

📌 3 Key Takeaways

🎯
Objective & Hypothesis: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.
📊
Key Findings & Data:Egg white cystatin was purified by using affinity chromatography on cm-papain-Sepharose.
💡
Conclusion & Impact: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.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

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.

Özet

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.

🔖

Cite This Article

AKPINAR, Ö. (2004). Rekombinant soyasistatinin saflaştırılması ve papaine ve balık proteazlarına karşı inhibitör aktivitesinin saptanması. Gaziosmanpaşa Üniversitesi Ziraat Fakültesi Dergisi, Cilt: 21 - Sayı: 1, 19–26. https://ajindex.com/articles/224702

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (12)

  1. Abe, K., Emöri, Y., Kondo, H., Suzuki, K. and Arai, S. 1987. Molecular cloning of a cysteine proteinase inhibitor of rice (Oryzacystatin). J. Biol. Chem. 262(35): 16793-16797.
  2. Abe, M., Abe, K., Iwabuchi, K., Domoto, C. and Arai S. 1994. Corn cystatin I expressed in Escherichia coli: investigation of its inhibitory profile and occurrence in corn kernels. J Biochem 116:488- 492. Abe, M., Abe, K., Domoto, C. and Arai, S. 1995. Two distinct species of corn cystatin in corn kernels. Biosci. Biotech. Biochem. 59(4): 756-758.
  3. An, H., Weerasinghe, V., Seymour, T.A., and Morrissey, M.T. 1994a. Cathepsin degradation of Pacific whiting surimi proteins. J Food Sci 5:1013-1017
  4. An, H., Seymour, T.A., Wu, J.W., and Morrissey MT. 1994b. Assay systems and characterization of Pacific whiting (Merluccius products) protease. J Food Sci 59:277-281.
  5. An, H., Peters, Y.M., and Seymour, T.A. 1996. Roles of endogenous enzyme in surimi gelation. Treds in Food Sci and Tech 7:321-327.
  6. Anastasi, A., Brown, M.A., Kembhavi, A.A., Nicklin, M.J.H., Sayers, C.A., Sunter, D.C., and Barrett, A.J. 1983. Cystatin, a protein inhibitor of cysteine proteinases. Biochem J 211:129-138.
  7. Arai, S., Watanabe, H., Kondo, H., Emori, Y., and Abe, K. 1991. Papain inhibitory activity of oryzacystatin, a rice seed cysteine proteinase inhibitor, depends on the central Gln-Val-Val- Ala-Gly region conserved among cystatin superfamily members. J Biochem 109:294-298.
  8. Axen, R., and Ernback, S. 1971. Chemical fixation of enzymes to cyanogen halide activated polysaccharide carriers. Eur J Biochem 18:351- 360. Barrett, A.J. 1981. Cystatin, the egg white inhibitor of cysteine proteinases. Methods in Enzymol 80:771-780.
  9. Barrett, A.J., and Kirschke, H. 1981. Cathepsin B, cathepsin, H, and cathepsin L. Methods in Enzymol 80:535-561.
  10. Barrett, A.J., Rawlings, N.D., Davies, M.E., Machleidt, W., Salvesan, G., and Turk, V. 1986. Cysteine proteinase inhibitors of cystatin superfamily. In: Barrett AJ, Salvesan G, editors. Proteinase Inhibitors The Netharlands Amsterdam: Elsevier Science Publishers B.V Bjork, I., Alriksson, E., and Ylinenejarvi, K. 1989. Kinetics binding of chicken cystatin to papain. Biochemistry. 28:1568-1573.
  11. Bode, W., Engh, R., Musil, D.J., Thiele, U., Huber, R., Karshikow, A., Brzin, J., Kos, J., and Turk, V. 1988. The 2.0 A X-ray crystal structure of chicken egg white cystatin and its possible mode of interaction with cysteine proteinases. EMBO J 7:2593-2599.
  12. Chen, G., Tang, S., Chen, C., and Jiang, S. 2000