Enrichment of myofibrillar proteins from beef muscle by a simple sequential method.

摘要

A great deal of attention has been paid to the most important proteins of the myofibrillar system, myosin, actin, titin and nebulin, because of their potential role in meat processing. Several methods from different sources have been reported for their isolation. However, most of them are complex, tedious and focus on the isolation of 1 protein at a time. In the present study, these 4 proteins were co-enriched simultaneously using a simple methodology. Myofibrillar proteins extracted from semitendinosus beef muscles were submitted to ammonium sulfate precipitation. Titin and nebulin were precipitated predominantly in the saturation range 40-60 g/l, whereas myosin heavy chain and actin were precipitated in the range 60-100 g/l. Denaturing PAGE and electroelution were employed for the separation and isolation of the proteins of interest from the corresponding salt fractions. The Western blot procedure was applied to detect and precisely identify the nebulin band.