Properties and some in vitro studies of 6-phosphogluconate dehydrogenase purified from the liver of Chalcalburnus tarichi, the only fish living in Lake Van's highly alkaline water (pH 9.8)
Properties and some in vitro studies of 6-phosphogluconate dehydrogenase purified from the liver of Chalcalburnus tarichi, the only fish living in Lake Van's highly alkaline water (pH 9.8)
In this study, some enzymology parameters and some antibiotics affecting the enzyme activity of 6- phosphogluconate dehydrogenase (6PGD) extracted from t the livers of fish from Lake Van (Chalcalburnus tarichi) were investigated because it is an important enzyme-producing NADPH, a reductive power, protecting the cell against the oxidative agents by producing reduced glutathione. The crude enzyme solution was obtained by using the affinity chromatography method. The native molecular weight of the enzyme is ∼90,000 ± 3000 Da and it is composed of two subunits with identical molecular weights of ∼46,000 Da as exhibited on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The pH and temperature for optimal conditions of 6PGD were about 8.5 and 40 ◦ C, respectively, and the enzyme showed optimal activity in 40 mM ammonium sulfate solution. In addition, in vitro effects of clindamycin phosphate, streptomycin sulfate, and lincomycin antibiotics on the enzyme activity were investigated.
___
- 1. Deyrolle, T. Rev. Mag. Zool. (Paris) 2ieme serie 1872, 23, 401405.
- 2. C¸ etinkaya, O. In Eastern Anatolia I. and II. Symposium on Fisheries, Erzurum, Turkey, 1996, pp. 7183.
- 3. Unal, G.; T¨urko˘glu, V.; O˘guz, A. R. ¨ Fish Physiol. Biochem. 2007, 33, 153165.
- 4. T¨urko˘glu, V.; Altun, M.; C¸ift¸ci, M. J. Physiol. Biochem. 2006, 62, 155161.
- 5. C¸ift¸ci, M.; T¨urko˘glu, V.; C¸ oban, T. A. J. Hazard. Materials 2007, 143, 415418.
- 6. Lehninger, A. L.; Nelson, D. L.; Cox, M. M. Lehninger Principles of Biochemistry; W.H. Freeman: New York, NY, USA, 2000.
- 7. Bianchi, D.; Bertrand, O.; Haupt, K.; Coello, N. Enzyme Microb. Technol. 2001, 28, 754759.
- 8. Bisswanger, H. Enzyme Kinetics: Principles and Methods; Wiley: Weinheim, Germany, 2008.
- 9. Chaplin, M. F.; Buche, C. Enzyme Technology; Cambridge University Press: New York, NY, USA, 1990.
- 10. Maglysh, S. S.; Gorbach, Z. V.; Ostrovskii, I. U. M. Biokhimiia 1982, 47, 20352041.
- 11. Villet, R. H.; Dalziel, K. Biochem. J. 1967, 104, 3839.
- 12. Somers, D. O.; Hajdu, J.; Adams, M. J. Protein Expr. Purif. 1991, 2, 385389.
- 13. Toews, M. L.; Kanji, M. L.; Carper, W. R. J. Biol. Chem. 1976, 251, 71277131.
- 14. G¨uler, M.; Kıvan¸c, M. R.; T¨urko˘glu, V.; Basi, Z.; Kıvrak, H. Bull. Environ. Contam. Toxicol. 2013, 9, 560564.
- 15. Prath, R. J.; Ernback, S. Nature 1967, 214, 13021304.
- 16. Cuatrecases, P.; Anfinsen, C. B. Methods in Enzymology; Academic Press: New York, NY, USA, 1971.
- 17. Sawa, Y.; Suzuki, K.; Ochiai, H. Agric. Biol. Chem. 1985, 49, 25432549.
- 18. Bridges, R. B.; Palumbo, M. P.; Wittenbreger, C. L. J. Biol. Chem. 1975, 250, 60936100.
- 19. Beydemir, B.; C¸ift¸ci, M.; Yılmaz, H.; K¨uvrevio˘glu, O. I. Turk J. Vet. Anim. Sci. 2004, 28, 707714.
- 20. Carne, A. Anal. Biochem. 1982, 121, 227229.
- 21. Weisz, K. S.; Schofield, P. J.; Edwards, M. R. J. Neurochem. 1985, 44, 510517.
- 22. Procsal, D.; Holten, D. Biochem. 1972, 11, 13101314.
- 23. Dyson, J. E.; DOrazio, R. E.; Hanson, W. L. T. J. Biol. Chem. 1973, 248, 54285435.
- 24. Pearse, B.; Rosemeyer, M. A. Eur. J.Biochem. 1974, 42, 213223.
- 25. Ninfali, P.; Orsenigo, T.; Barociani, S. R. Prep. Biochem. Biotechnol. 1990, 20, 297309.
- 26. Aky¨uz, M.; Erat, M.; C¸ift¸ci, M.; G¨um¨u¸stekin, K.; Bakan, N. J. Enzyme Inhib. Med. Chem. 2013, 19, 361365.
- 27. Bradford, M. M. Anal. Biochem. 1976, 72, 248251.
- 28. Beutler, E. Red Cell Metabolism; Grune & Stratton: New York, NY, USA, 1984.
- 29. Laemmli, D. K. Nature 1970, 227, 680683.
- 30. Rosemeyer, M. N. Cell. Biochem. Func. 1987, 5, 7995.