Optimization of Chromatographic Criterion to Recover Indian White Shrimp (Fenneropenaeus indicus) Alkaline Phosphatase from Hepatopancreas.

ABSTRACT

Commercially important alkaline phosphatase can be recovered from the hepatopancreatic tissue of Indian white shrimp with optimum yield, activity, and purity by carefully designing the criterions of ion exchange chromatographic purification. The tissues were homogenized, clarified, and concentrated. Concentrated homogenate was purified with optimum yield, activity and purity by optimising criterions of chromatography such as pH and ionic strength of the binding buffer, and ionic gradient and the flow rate of the elution buffer in DEAE-cellulose column. The enzyme was optimally bound to the column at pH 8.4 and ionic strength 0.1 M NaCl at flow rate of 1mL/min, and eluted with high resolution at ionic gradient of 0.10-0.35 M NaCl in 25 min at a flow rate of 1.5mL/min, and hence is the optimum criterions of chromatographic purification.