Effective removal of insolubles from brown shrimp hepatopancreatic homogenate during primary stages of alkaline phosphatase recovery

Abstract

Krishna Prasad Nooralabettu

Background and Objectives: Commercially important alkaline phosphatase of novel characteristics can be isolated from the hepatopancreatic tissues of Brown shrimp (Metapenaeus monoceros). However, interference of insoluble released during the homogenisation may interfere with subsequent recovery steps. Hence, present study is aimed at designing an effective insoluble removal method to isolate alkaline phosphatase with optimum yield and activity. Materials and Methods: Hepatopancreatic tissues of the shrimp was homogenized at 3,000 rpm for 10 min at 4oC in a homogenizer and the tissue homogenates were centrifuged at Relative Centrifugal Force of 67.2, 1681.1, 6724.3, 15124.8 or 26897.4 × g for 5, 10, 15, or 20 min at 4°C. Results: Relative Centrifugal Force of 1681.1×g for 5 min at 4°C successfully clarified 90.28±1.55% of the insolubles. The forces below this level even upto 20 min removed only up to 56.57±0.66% of the total solids and 7.90±0.19% of the total lipids from the homogenate. Force and time above RCF of 1681.1×g for 5 min, specific activity was increased by 4.18±0.38 folds due to the loss of total protein, but upto 60.48±2.61% of alkaline phosphatase activity was lost. Conclusion: Hence, the force of 1681.1 × g and resident time of 5 min in the centrifuge is an efficient clarification method of tissue homogenate