Simultaneous purification of superoxide dismutase, catalase and hemoglobin from bovine erythrocyte lysate with PEG 600 as chaperon / 生物工程学报

ABSTRACT

Superoxide dismutase, catalase and hemoglobin were purified simultaneously from the same batch of bovine erythrocyte lysate. The process involves an initial anion exchange chromatography, followed by a hydrophobic interaction chromatography and gel filtration chromatography. 0.75% polyethylene glycol 600 was added as a purification chaperon before the anion exchange chromatography. The hemoglobin fraction passed through the ion exchange column without being retained. The superoxide dismutase and catalase were adsorbed by the column and were eluted separately during elution. The two eluted fractions containing crude superboxide dismutase and catalase were further purified with hydrophobic interaction chromatography and gel filtration chromatography in sequence. The specific activities of superoxide dismutase and catalase were 15932u/mg and 65918u/mg, respectively. SDS-polyacrylamide gel electrophoresis and gel filtration chromatography were used to analyze the purity of the proteins. The purity of superoxide dismutase, catalase and hemoglobin were 77.6%, 81.9% and 99.9%, respectively. The total recoveries for superoxide dismutase, catalase and hemoglobin were 47.4%, 29.6% and 88.7%, respectively.