Research Article | Volume: 4, Issue: 1, Jan-Feb, 2016

Properties of phenoloxidase from haemocytes of Scylla tranquebarica

A. S. Sabu K. P. Jithendran C Gopal   

Open Access   

Published:  Feb 19, 2016

DOI: 10.7324/JABB.2016.40109

The presence of phenoloxidase (PO) activity in the haemocytes of Scylla tranquebarica was electrophoretically and spectrophotometrically studied. Majority of the enzyme was located as proenzyme, prophenoloxidase (proPO) in the haemocytes. The enzyme prefers L-dihydroxyphenylalanine (L-dopa) as its substrate than phenol and is optimally active at pH 8.0. Besides trypsin, the proPO was also activated by both Gram positive and Gram negative microbes in vitro while, chemicals such as sodium azide, thiourea and EDTA significantly inhibited the enzyme expression. The protein needs considerable levels of divalent cations like calcium (20 mM as CaCl2) or magnesium (20–50 mM as MgCl2) for its activity. The gel filtration chromatography of haemocyte lysate supernatant showed a single major peak of protein having PO activity. Electrophoresis of purified PO by native PAGE revealed a single prominent band of approximately 167.2 kDa which was further resolved to three bands having molecular mass of approximately 77.1, 56.9 and 30.2 kDa respectively, on SDS-PAGE.

Keyword:     Crab Scylla tranquebarica Haemocyte Prophenoloxidase Phenoloxidase.


Sabu AS, Jithendran KP, Gopal C. Properties of phenoloxidase from haemocytes of Scylla tranquebarica. J App Biol Biotech. 2016; 4 (01): 047-052. DOI: 10.7324/JABB.2016.40109

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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1. Bachere E. Shrimp immunity and disease control. Aquaculture. 2000; 191: 3-11.

2. Destoumieux D, Muñoz M, Cosseau C, Rodriguez J, Bulet P, Comps M, Bachere E. Penaeidins, antimicrobial peptides with chitin-binding activity, are produced and stored in shrimp granulocytes and released after microbial challenge. Journal of cell science. 2000; 113: 461-469.

3. Sritunyalucksana K, Söderhäll K. The proPO and clotting system in crustaceans. Aquaculture. 2000; 191: 53-69.

4. Söderhäll K, Cerenius L. Role of the prophenoloxidase-activating system in invertebrate immunity. Current Opininon Immunology. 1998; 10: 23-28.

5. Sung HH, Chang HJ, Her CH, Chang JC, Song YL. Phenoloxidase activity of hemocytes derived from Penaeus monodon and Macrobrachium rosenbergii. Journal of Invertebrate Pathology. 1998; 71: 26-33.

6. Houton C, Williams JA, Hawkins LE. The effects of a live in vivo pathogenic infection on aspects of the immunocompetence of the common shore crab, Carcinus maenas (L.). Journal of Experimental Marine Biology and Ecology. 1997; 211: 115-128.

7. Gollas-Galván T, Hernández-López J, Vargas-Albores F. Prophenoloxidase from brown shrimp (Penaeus californiensis) hemocytes. Comparative Biochemistry and Physiology. 1999; 122B: 77-82.

8. Mangkalanan S, Sanguanrat P, Utairangsri T, Sritunyalucksana K, Krittanai C. Characterization of the circulating hemocytes in mud crab (Scylla olivacea) revealed phenoloxidase activity. Devlopmental and Comparative Immunology. 2014; 44: 116-123.

9. Söderhäll K, Smith VJ. Separation of the hemocyte population of Carcinus maenas and other marine decapods and prophenoloxidase distribution. Developmental and Comparative Immunology. 1983; 7: 229-239.

10. Alsina M, Blanch AR. Improvement and uptake of a set of keys for the biochemical identificatyion of Vibrio species. Journal of Applied Bacteriology. 1994; 77: 719-721.

11. Laemmli U.K. 1970. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680-685.

12. Lowry OH. Rosebrough NL, Farr AL, Randall RJ. Protein measurement with Folin phenol reagent. Journal of Biological Chemistry. 1951; 193: 265–275.

13. Söderhäll K, Cerenius L. Role of the prophenoloxidase-activating system in invertebrate immunity. Current Opinion Immunology. 1998; 10: 23-28.

14. Söderhäll K, Cerenius L. Crustacean immunity. Annual Review of Fish Diseases. 1998; 2: 3-23.

15. Wilson K. Protein and enzyme techniques. In: Wlison K, Walker J, editors. Practical Biochemistry. Principles and Techniques, Cambridge University Press; Cambridge; 1995, p. 162-226.

16. Liu H1, Jiravanichpaisal P, Cerenius L, Lee BL, Söderhäll I, Söderhäll K. Phenoloxidase is an important component of the defense against Aeromonas hydrophila Infection in a crustacean, Pacifastacus leniusculus. Journal of Biological Chemistry. 2007; 282: 33593-33598.

17. Leonard C, Ratcliffe NA, Rowley AF. The role of prophenoloxidase activation in non-self recognition and phagocytosis by insect blood cells. Journal of Insect Physiology. 1985; 31: 789–799.

18. Hernández-López J, Gollas-Galván T, Vargas-Albores F. Activation of the prophenoloxidase system of the brown shrimp (Penaeus californiensis Holmes). Comparative Biochemistry and Physiology. 1996; 113C: 61-66.

19. Sartoris FJ, Põrtner HO. Increased concentrations of haemolymph Mg2+ protect intracellular pH and ATP levels during temperature stress and anoxia in the common shrimp Crangon crangon. Journal of Experimental Biology. 1997; 200: 785-792.

20. Aspán A, Söderhäll K. Purification of prophenoloxidase from crayfish blood cells, and its activation by an endogenous serine proteinase. Insect Biochemistry. 1991; 21: 363-373.

21. Lanz H, Hernández S, Garrido-Guerrero E, Tsutsumi V, Aréchiga H. Prophenoloxidase system activation in the crayfish Procambarus clarkii. Developmental and Comparative Immunology. 1993; 17: 399-406.

22. Liu G, Yang L, Fan T, Cong R, Tang Z, Sun W, Meng X, Zhu L. Purification and characterization of phenoloxidase from crab Charybdis japonica. Fish and Shellfish Immunology. 2006; 20: 47-57.

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