Optimized method for myoglobin purification and the study of physical characterization

Myoglobin is a major hemoprotein and the principal oxygen reservoir in the muscle tissues of liveanimals. There are different methods for isolation and purification of myoglobin but in this study a simpleand rapid procedure was developed to isolate and purify myoglobin from animal muscles. This methodinvolved fractional precipitation of a crude myoglobin extract with ammonium sulfate and purificationwith a single chromatographic step on a Sephadex G-100 column. Phosphate buffer and Tris-HCl bufferwere used for extraction and purification. The purity of the myoglobin preparations was confirmed bySDS-polyacrylamide gel electrophoresis. Sephadex G-100 chromatography of crude myoglobin extractsof meat produced similar elution profiles when monitored at 412 and 525 nm that was similar to sigmamyoglobin. Two kinds of different myoglobin that extracted by this method were studied by fluorescence,circular dichroism and uv-visible spectroscopic methods.

کلیدواژه ها:

Myoglobin ، Chromatography ، Sephadex G-100 ، Fluorescence