D-Lactate Dehydrogenase (Cytochrome) from Saccharomyces cerevisiae Purification by Fast Protein Liquid Chromatography

• Autorzy: Pohanka M. , Zbořil P. , Pikula J.
• Języki publikacji: EN

Abstrakty

EN

D-Lactate dehydrogenase (cytochrome) was isolated from baker´s yeasts (Saccharomyces cerevisiae) using fast protein liquid chromatography. Mitochondria from disrupted cells were separated and enzymes released by Triton X-100. Purification was performed by fast protein liquid chromatography on hydroxylapatite and in another separate experiment on AH-Sepharose with covalently bound 4-hydroxy-acyanocinnamic acid (SHCA). Although purification on hydroxylapatite provided approachable samples with activities for only D-lactic acid, chromatography on SHCA seemed to be better because a purified fraction of enzyme with the specific activity of 3.38 nkat/mg was obtained. Another problem considered was the usefulness of the combination of chromatography on hydroxylapatite and SHCA providing a fraction of D-lactic acid with specific activity of 5.71 nkat/mg and no cross reactivity for L-lactic acid.

Słowa kluczowe

EN

baker's yeast; mitochondria; d-lactic acid; isolation

• Wydawca: Polskie Towarzystwo Chemiczne
• Czasopismo: Polish Journal of Chemistry
• Rocznik: 2009
• Tom: Vol. 83, nr 3
• Strony: 415--420
• Opis fizyczny: Bibliogr. 16 poz., rys.

Twórcy

autor

Pohanka M.

autor

Zbořil P.

autor

Pikula J.

• Centre of Advanced Studies and Department of Toxicology, Faculty of Military Health Sciences, University of Defence, Trebesska 1575, 500 01 Hradec Kralove, Czech Republic Tel: [phone],,

Bibliografia

• 1.    Garvie E.I., Microbiol. Rev., 44, 1905 (1980).
• 2.    Bergmeyer H.U., Methods of enzymatic analysis, p. 582, VCH, Weinheim (1990).
• 3.    Montagne M. and Marty J., Anal. Chim. Acta, 315, 297 (1995).
• 4.    Girotti S., Muratori M., Fini R, Ferri E.N., Carrea G., Koran M. and Rauch R, Eur. FoodRes. Technol.,210,216(2000).
• 5.    Haddock B.A. and Jones C.W., Bacteriol Rev., 41, 47 (1977).
• 6.    Pohanka M. and Zboril R, Food Technol. Biotechnol., 46, 107 (2008).
• 7.    Gregolin C. and Singer T.R, Ann. N.Y. Acad. Sci., 94, 780 (1961).
• 8.    Gregolin C. and Singer T.R, Biochim. Biophys. Acta, 67, 201 (1963).
• 9.   Ogata M., Arihara K. and Yagi T., J. Biochem., (Tokyo), 89, 1423 (1981).
• 10. Olson S.T. and Massey V., Biochemistry, 18, 4714 (1979).
• 11. Reed D. W. and Hartzell R, J. Bacteriol., 181, 4580 (1999).
• 12.   Lodi T., O'Connor D., Goffrini R and Ferrero L, Mol. Gen. Genet., 244, 622 (1994).
• 13.   Avers C.J., Szabo A. and Price C.A., J. Bacteriol., 100, 1044 (1969).
• 14.   Lodi T. and Ferrero L, Mol. Gen. Genet., 238, 315 (1993).
• 15.   Lodi T., Alberti A., Guiard B. and Ferrero L, Mol. Gen. Genet., 262, 623 (1999).
• 16.   Lowry O.H., Roselbrough N.J., Farr A.L. and Randall R.J., J. Biol. Chem., 193, 265 (1951).