Publication details
D-Lactate Dehydrogenase (Cytochrome) from Saccharomyces cerevisiae Purification by Fast Protein Liquid Chromatography
| Authors | |
|---|---|
| Year of publication | 2009 |
| Type | Article in Periodical |
| Magazine / Source | Polish journal of chemistry |
| MU Faculty or unit | |
| Citation | |
| web | http://www.ichf.edu.pl/pjch/pj-2009.htm |
| Keywords | baker's yeast; mitochondria; d-lactic acid; isolation |
| Description | 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-alpha cyanocinnamic acid (SHCA). Although purification on hydtoxylapatite 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. |