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1 Polish Journal of Chemistry

1 2009

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

Pohanka M., Zbořil P., Pikula J.

Polish Journal of Chemistry

2009

Vol. 83, nr 3

415-420

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.