PURIFICATION OF CYTOCHROME C PEROXIDASE DERIVED FROM YEAST

摘要

PURPOSE: To purify the subject enzyme in high purity in a short time by fractionating and precipitating a crude protein extract containing cytochrome C peroxidase obtained from a yeast lysate and treating the precipitate by specific two kinds of chromatographies, respectively. ;CONSTITUTION: First, a lysate of a yeast (generally Saccharomyces cerevisiae) is preferably extracted with a buffer solution having pH≥5 to give a crude protein extract of cytochrome C peroxidase (CCP for short). Then the extract is fractionated and precipitated with highly concentrated ammonium sulfate having preferably ≤80 wt./vol.%, the CCP-containing precipitate is treated by an anion exchange chromatography to give partially pure CCP. Finally the CCP is treated by phosphate affinity chromatography using hydroxyapatite to give the objective CCP.;COPYRIGHT: (C)1990,JPO