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Purification and characterization of prophenoloxidase from cotton bollworm, H elicoverpa armigera
Author(s) -
Goudru Hanumanth G.,
Kumar Sathish,
Jayalakshmi Senigala K.,
Ballal Chandish R.,
Sharma Hari C.,
Sreeramulu Kuruba
Publication year - 2013
Publication title -
entomological research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.421
H-Index - 20
eISSN - 1748-5967
pISSN - 1738-2297
DOI - 10.1111/1748-5967.12002
Subject(s) - prophenoloxidase , biology , biochemistry , dithiothreitol , enzyme , kojic acid , tyrosinase , receptor , innate immune system
Phenoloxidases are oxidative enzymes, which play an important role in both cell mediated and humoral immunity. Purification and biochemical characterization of prophenoloxidase from cotton bollworm, H elicoverpa armigera ( H übner) were carried out to study its biochemical properties. Prophenoloxidase consists of a single polypeptide chain with a relative molecular weight of 85 k D a as determined by SDS – PAGE , MALDI – TOF MS and LC – ESI MS . After the final step, the enzyme showed 71.7 fold of purification with a recovery of 49.2%. Purified prophenoloxidase showed high specific activity and homology with phenoloxidase subunit‐1 of B ombyx mori and the conserved regions of copper binding ( B ) site of phenoloxidase. Purified prophenoloxidase has pH optima of 6.8 and has high catalytic efficiency towards the dopamine as a substrate in comparison to catechol and L‐D opa. The PO activity was strongly inhibited by phenylthiourea, thiourea, dithiothreitol and kojic acid.