Abstract
A procedure for isolation and purification of aspartate aminotransferase from wheat grain includes chromatography on DEAE cellulose, acidification–alkalization, precipitation with protamine sulfate, fractionation with ammonium sulfate, and chromatography on hydroxyapatite. The yield of protein was 27% with 95% purity. Crystals of the enzyme (0.05 × 0.025 × 0.015 mm3) were obtained from ammonium sulfate solution.
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REFERENCES
Christen, P., and Metzler, D. E. (1985) Transaminases, J. Wiley and Sons, New York.
Izard, T., Fol, B., Pauptit, R. A., and Jansonius, J. K. (1980) J. Mol. Biol., 215, 341–344.
Malashkevich, V. N., Strokopytov, B. V., Borisov, V. V., Dauter, Z., Wilson, K. S., and Torchinsky, Yu. M. (1995) J. Mol. Biol., 247, 111–124.
Kochkina, V. M., Korolev, S. V., Midor, V. I., Wilson, D., Kvocho, F. A., and Kuzin, A. P. (1995) Mol. Biol. (Moscow), 28, 333–341.
Givan, C. V. (1980) Aminotransferases in Higher Plants. The Biochemistry of Plants (Miflin, B. J., ed.) Vol. 5, Academic Press, New York, pp. 329–359.
Jenkins, W. T., Yphantis, D. A., and Sizer, J. W. (1957) J. Biol. Chem., 234, 51–57.
Karmen, A. (1955) J. Clin. Invest., 34, 131–133.
Laemmli, U. K. (1970) Nature, 227, 680–695.
Salemme, F. R. (1972) Arch. Biochem. Biophys., 151, 533–539.
McPherson, A. (1982) Preparation and Analysis of Protein Crystals, J. Wiley and Sons, New York, pp. 119–121.
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Kochkina, V.M. Isolation, Purification, and Crystallization of Aspartate Aminotransferase from Wheat Grain. Biochemistry (Moscow) 69, 897–900 (2004). https://doi.org/10.1023/B:BIRY.0000040222.03147.01
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DOI: https://doi.org/10.1023/B:BIRY.0000040222.03147.01