ROLE OF PROPHENOLOXIDASE (PROPO) IN SILKWORM IMMUNITY- DETERMINATION OF PHENOL OXIDASE (PO) ACTIVITY
|G.Sairamalinga Reddy1., B.Venkatappa2., P.Seshapani3., D.Jayasimha Rayalu4*.,.B.Janardhan5
|Corresponding Author: D.Jayasimha Rayalu, Department of Biotechnology, DR.Rayaluâs Biotech PVT.LTD, Himayat nagar, Hyderabad, AP,INDIA, E-mail: [email protected]|
|Received: 15 March 2012 Accepted: 23 March 2012|
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A fraction of hemolymph from healthy and bacteria inducted silkworms were collected and total protein content was estimated. Prophenoloxidase in hemolymph of bacteria infected silkworm (Bombyx mori) was partially purified and migrated as two bands in SDS-PAGE system, where as prophenoloxidase band is completely absent in cuticular protein sample of healthy silkworm. Extraction and characterization of pro-phenol oxidase cascade from healthily and bacteria included silkworms was used to determined Phenol oxidase activity. Effect of various sugars on development of phenoloxidase activity was examined. A cane sugar factor (CSF) seems responsible for retention of prophenoloxidase in Cane Sugar-Hemolymph. Microscopic studies revealed that cell clusters or aggregates showed a regular pattern. The results shown that a single protein having molecular mass of 18.5 kDa. These results have clear evidence that upon stimulation by LPS the levels of PEA derived peptide i.e peptide B the 13kDa fragment is significantly increased. Phenoloxidase activity incresed with time when the prophenoloxidase fraction was incubated with the activity cascade (AC) fraction that had been previously treated with Ca² . Presently results clearly demonstrate that cuticular phenoloxidase is truly is Ã¢ÂÂº Zymogen form which is activated through a limited proteolosis.