A Study of Purification Profile and Molecular Weight of Fish Visceral Protease Precipitated With Different Precipitating Agents
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Date
2014
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Abstract
The study was aimed at managing the vast amounts of wastes of the redfish Lutjanus sebea to isolate an
industrially useful enzyme-protease. The visceral organ waste.s of fish were collected, homogenized with
Tris-HCI buffer and precipitated with varying concentrations of ammonium sulphate, aeetone and ethanol
and was then purified by dialysis and Sephadex G-lOO column chromatography. The purification profile
(the protein content, protease activity, specific activity, purification fold, and recovery %) were studied in
the crude and partially purified samples. The results .showed that the activity was maximum when
precipitated with 40-60% ammonium sulphate, 100% acetone and 40% ethanol. The fractions with the
highest specific activity were selected for further purification by dialysis and Sephadex G-lOO column
chromatography. The molecular weight of the isolated enzyme was determined by SDS-PAGE and Native
PAGE and was to he found 27 kDa. The presence of protease was confirmed by zymography.