Kathyayani Puttige, Krishna Prasad Nooralabettu
Abstract:
Hepatopancreatic tissues of shrimps were homogenized at various pestle speed and time intervals at a constant jerk of 15 passes/min on a narrow space using 0.1 M Tris-HCl buffer of pH 8.4 at 4°C. Release of proteins and alkaline phosphatase were determined by estimating the quantity of protein and alkaline phosphatase activity in each homogenate at the end of the experiments. Optimum condition for the homogenization of the hepatopancreatic cells was achieved at operating speeds of 3,000 rpm for 10 min in the homogenizer. Increase in pestle speed beyond 3,000 rpm for 10 min have no effect on the release of protein. Increasing the disruption speed above 3,000 rpm decreases the specific activity of alkaline phosphatase release, while extending the time of homogenization beyond 10 min at 3,000 rpm or the speed of pestle beyond 3,000 rpm for 10 min caused shear damage to the enzyme.
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Keywords:
alkaline phosphatase, shrimp, cell disruption, homogenization, homogeniser
