Partial purification and some properties of aspartate aminotransferase from the mantle tissue of Mytilus galloprovincialis Lam

Year 2014, Volume: 44 Issue: 1, 1 - 10, 29.01.2015

Özlem Dönmez Yurtpınar Tuğba Yılmaz Özden Ayşe Can

https://izlik.org/JA97RG75LK

Abstract

Aspartate aminotransferase (E.C.2.6.1.1; AST), is a pyridoxal phosphate dependent enzyme that occurs in virtually all organisms and plays a key role in intermediary nitrogen metabolism. Although AST was purified from a variety of plant and animal sources, it has not been purified previously from mantle tissue of Mytilus galloprovincialis Lam. In the present study we have partially purified AST from the mantle tissue of M. galloprovincialis and investigated some kinetic properties of the enzyme. The partially purified enzyme showed three protein and a single activity band in polyacrylamide gel electrophoresis. It was found that the enzyme exhibited maximum activity at 15ºC and 35ºC and that the activity was decreased at 40ºC and totally lost at 55ºC. AST activity was maximum at pH 7.4 in Tris-HCl buffer. Km values of AST for aspartate and 2-oxoglutarate were 1.64 mM and 2.2x10-2 mM, respectively, and Vmax values for the same substrates were 0.12 U/mL and 0.168 U/mL, respectively

Keywords

aspartate aminotransferase , Mytilus galloprovincialis Lam. , mussel , enzyme purification

Partial purification and some properties of aspartate aminotransferase from the mantle tissue of Mytilus galloprovincialis Lam

https://izlik.org/JA97RG75LK

Abstract

References

• 1. Christen, P., Metzler, D.E. (eds.). Transaminases, p. 643, Wiley, New York (1985).
• -Ref: Hayashi, H. Recent topics in pyridoxal-5’-phosphate enzyme studies. Ann. Rev. Biochem., 59, 87-110 (1990).
• 2. Orlacchio, A., Campos-Cavieres, M., Pashev, I., Munn, E.A. Some kinetic and other properties of the isoenzymes of aspartate aminotransferase isolated from sheep liver. Biochem. J., 177, 583-593 (1979).
• 3. Akev, N., Can, A., Yanardağ, R. Lentil seed aspartate aminotransferase isoenzymes. I. Isolation and partial purification. İstanbul Ecz. Fak. Mec., 31, 1-9 (1995).
• 4. Öner, N., Can, A., Aydeğer, N., Tütem, E. Isolation and purification of two aspartate aminotransferase isoenzymes from soybean seeds. Acta Pharm. Turcica, 30, 43-48 (1988).
• 5. Lis, H. Purification of glutamic-aspartic transaminase. Biochim. Biophys. Acta, 28, 191-197 (1958).
• 6. Bishop, S.H., Ellis, L.L., Burcham, J.M. Amino acid metabolism. In: Hochanchka, P.W., Wilbur, K.M. (eds.). The Mollusca, Metabolic Biochemistry and Molecular Biomechanics, Vol. 1, p. 243-327, Academic Press Inc, New York (1983). -Ref: Mc Cormic, A., Paynter, K.T., Brodey, M.M., Bishop, S.H. Aspartate aminotranferases from ribbed mussel gill tissue: Reactivity with β-L-cysteine sulfinic acid and other properties. Comp. Biochem. Phys., 84B, 163-166 (1986).
• 7. Goromosova, S.A., Tamozhnyaya, V.A. Seasonal dynamics of ferments of transamination in tissues of Black Sea mussels. Biology Marine, 2, 62-68 (1980).
• 8. Pastore, M., De Lisi, A., Biandolino, F., Marzulli, D. Enzymatic activity in Mytilus galloprovincialis (Mollusca: Bivalvia) digestive gland. J. Mar. Biol. Ass. U.K., 81, 701-702 (2001).
• 9. Tiselius, A., Hjertén, S., Levin, Ö. Protein chromatography on calcium phosphate columns. Arch. Biochem. Biophys., 65, 132-155 (1956).
• 10. Lowry, O.H., Rosenbrough, N.J., Farr, A.L., Randall, R. Protein measurment with the folin phenol reagent. J. Biol. Chem., 193, 265-275 (1951).
• 11. Warburg, O., Christian, W. Isolierung und kristallinsation des gärungsferments enolase. Biochem. Z., 310, 384-421 (1941).
• 12. Karmen, A. A note on the spectrophotometric assay of glutamic-oxalacetic transaminase in human blood serum. J. Clin. Invest., 34, 131-133 (1955).
• 13. Rej, R., Bretaudiere, J.P., Graffunder, B. Measurment of aspartate aminotransferases isoenzymes: Six procedures compared. Clin. Chem., 27, 535-542 (1981).
• 14. Frankel, S., Reitman, S., Sonnenwirth, A.C. Gradwohl’s Clinical Laboratory Methods and Diagnosis, Vol.1, p. 126, The C.V. Mosby Company, Saint Louis (1970).
• 15. Ornstein, L. Disc Electrophoresis-I. Background and Theory. Ann. N.Y. Acad. Sci., 121, 321-349 (1964).
• 16. Davis, BJ. Disc Electrophoresis-II. Method and application to human serum proteins. Ann. N.Y. Acad. Sci., 121, 404-427 (1964).
• 17. Schwartz, M.K., Nisselbaum, J.S., Bodansky, O. Procedure for staining zones of activity of glutamic oxalacetic transaminase following electrophoresis with starch gel. Am. J. Clin. Pathol., 40, 103-106 (1963).
• 18. Can, A., Akev, N., Askhasi, A., Partial purification and characterization of aspartate aminotransferase from the hepatopancreas of the mussel Mytilus galloprovincialis Lam. İstanbul Ecz. Fak. Mec., 33, 37-48 (2000).
• 19. Paynter, K.T., Hoffmann, R.J., Ellis, L.L., Bishop, S.H. Partial characterization of the cytosolic and mitochondrial aspartate aminotransferase from ribbed mussel gill tissue. J. Exp. Zool., 231, 185-197 (1984).
• 20. Jenkins, W.W.T., Ypantis, D.A., Sizer, I.W. Glutamic aspartic transaminase. 1.Assay, purification and general properties. J. Biol. Chem., 234, 51-57 (1959).
• 21. Öner, N., Aydeğer, N., Can, A. Some properties of aspartate aminotransferase isoenzymes from soybean seeds. Acta Pharm. Turcica, 31, 27-32 (1989).
• 22. Sauvage, F.X., Romieu, C.G., Flanzy, C., Robin, J.P. Aminotransferases in grapes. Isolation and characterization of aspartate aminotransferase. Am. J. Enol. Vitic., 42, 209-218 (1991).
• 23. Can. A., Akev, N. Lentil seeds aspartate aminotransferase isoenzymes. II. Characterization. Acta Pharm. Turcica, 37, 100-104 (1995).
• 24. Berkkan, H. Midye amilazlarının saflaştırılması ve başlıca kinetik özelliklerinin incelenmesi. İstanbul Üniv. Ecz. Fak. Biokimya Anabilim Dalı, Doçentlik Tezi (1982).
• 25. McCormic, A., Paynter, K.T., Brodey, M.M., Bishop, SH. Aspartate aminotransferases from ribbed mussel gill tissue: reactivity with β-L-cysteine sulfinic acid and other properties. Comp. Biochem. Physiol., 84B, 163-166 (1986).
• 26. Nisselbaum, J.S., Bodansky, O. Kinetic and electrophoretic properties of the isozymes of aspartate aminotransferase from pig heart. J. Biol. Chem., 241, 2661-2664 (1966).