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Rekombinant Soyasistatinin Saflaştırılması ve Papaine ve Balık Proteazlarına Karşı İnhibitor Aktivitesinin Saptanması

Purification and Determination of Inhibitory Activity of Recombinant Soyacystatin Against Papain and Fish Protease

Journal Name:

  • Gaziosmanpaşa Üniversitesi Ziraat Fakültesi Dergisi

Publication Year:

  • 2004

Key Words:

Keywords (Original Language):

Author NameUniversity of AuthorFaculty of Author
Abstract (2. Language): 
Recombinant (r-) soyacystatin was characterized for its inhibitory activity against papain and compared to egg white cystatin. 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 m fish muscle.
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Abstract (Original Language): 
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 afınite kromatografisi kullanılarak saflaştırılmıştır. 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-Soyasıstatın araştırmada kullanılan balık kasındaki otolitik aktivitenin %90 nu inhibe etmiştir.
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  • Turkish

REFERENCES

References: 

Abe, K., Emori, 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.
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 com cystatin in com kernels. Biosci. Biotech. Biochem. 59(4): 756-758.
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
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.
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.
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.
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.
Axen, R., and Emback, 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.
Barrett, A.J., and Kirschke, H. 1981. Cathepsin B, cathepsin, H, and cathepsin L. Methods in Enzymol 80:535-561.
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, L, Alriksson, E., and Ylinenejarvi, K. 1989.
Kinetics binding of chicken cystatin to papain. Biochemistry. 28:1568-1573.
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.
Chen, G., Tang, S., Chen, C, and Jiang, S. 2000.
Overexpression of the soluble from of chicken cystatin in Escherichia coli and its purification. J Agric Food Chem 48:2602-2607.
Fossum, K., and Whitaker, J.R. 1968. Ficin and papain inhibitor from chicken egg white. Arch Biochem Biophys 125:367-375.
Gounaris, A.D., Brown, M.A., and Barrett, A.J. 1984.
Human plasma a-cysteine proteinase inhibitor. Purification by affinity chromatography, characterization and isolation of an active fragment. Biochem J 221: 445-452
Hsieh, J.F., Tsai, G.J., and Jiang, S.T. 2002. Improvement of hairtail surimi gel properties by NADPH-sulfite reductase, recombinant cystatin, and microbial transglutaminase. J Food Sci 67:31 52¬3158.
Izquierdo-Pulido, M.L., Haard, T.A., Hmg, J., and Haard.
N.F. 1994. Oryzacystatin and other proteinase inhibitors in rice grain: potential use as a fish processing aid. J Agric Food Chem 42:616-622
Jiang, ST., Tzeng, S.S., Wu, W.T., and Chen, G.H. 2002.
Enhanced expression of chicken cystatin as a thioredoxin fusion from in Escherichia coli AS494(DE3)pLysS and its effect on the prevention of surimi gel softening. J Agric Food Chem 50:3731-3737.
Kang, I.S., and Lanier, T.C. 1999. Bovine plasma protein function in surimi gelation compared with cysteine protease inhibitors. J Food Sci 64:842¬846.
Koiwa, H., Shade, R.E., Zhu-Salzman, K., Subramanian, L., Murdock, L.L., Nielsen, S.S., Bressan, R.A., and Hasegawa, P.M. 1998. Phage display selection can differentiate activity of soybean cystatins. Plant J 13:101-109.
Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:860-865.
Lee, J.J., Tzeng, S.S., Wu, J., and Jiang, S.T. 2000. Inhibition of thermal degradation of mackerel surimi by pig plasma protein and L-kininogen. J Food Sci 65:1124-1129.
Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, RJ. 1951. Protein measurement with Folin phenol Teagent. J Biol Chem 193: 256-946.
Morrissey, M.T., Wu, J.W., Lin, D., and An, H. 1993.
Protease inhibitor effects on torsion measurments and autolysis of Pacific whiting surimi. J Food Sci 5:1050-1054.
Nicklin, M.J.H. and Barrett, A.J. 1984. Inhibition of cysteine proteinases and dipeptidyl peptidase I by egg white cystatin. Biochem. J 223:254-253.
Robyt, F.J., and White, B.J. 1990. Biochemical Techniques Theory and Practice. Prospect Heights: Waveland Press.
Schagger, H., and Jagow, G.V.1987. Tricine-sodium dodecyl sulfate-polyacrylamide ge!
electrophoresis for separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166:368-379.
Schwabe, C, Anastasi, A., Crow, H., McDonald, J.K., and Barrett, A.J. 1984. Cystatin: amino acid sequence and possible secondary structure. Biochem J 217:813-817.
25
Ö.AKPINAR
Turk, V., Brzin, J., Longer, M., Ritonja, A., and Eropkin, M. 1983. Protein inhibitors of cysteine proteinases: III amino-acid sequences of cystatin from chicken egg white. Hoppe-Seyler's Z Physiol Chem 364:1487-1496.
Turk, B., Turk, V., and Turk, D. 1997. Structural and functional aspects of papain-like cysteine proteinases and their protein inhibitors. Biol Chem Hoppe-Seyler 378:141-150.
Tzeng, S.S., Chen, G.H., Chung, Y.C., and Jiang, S.T.
2001. Expression of soluble form carp (Cyprinus carpio) ovarian cystatin in Escherichia coli and its purification. J Agric Food Chem 49:4224¬4230.
Wasson, D.H., Reppond, K.D., and French, J.S. 1992.
Characterization of a heat stable protease from arrowtooth flounder. Atheresthes stomias. J Aquat Food Prod Technol 167:167-182.
Visessanguan, W., Menino, A.F., Kim, S.M., and An, H.
2001. Cathespsin L: Predominat heat-activated proteinase in Arrowtooth Flounder muscle. J Agric Food Chem 49:2633-2640.
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