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The molecular characterization of a catalase from C hinese mitten crab E riocheir sinensis
Author(s) -
Wang M.,
Wang L.,
Zhou Z.,
Gao Y.,
Wang L.,
Shi X.,
Gai Y.,
Mu C.,
Song L.
Publication year - 2013
Publication title -
international journal of immunogenetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.41
H-Index - 47
eISSN - 1744-313X
pISSN - 1744-3121
DOI - 10.1111/iji.12019
Subject(s) - eriocheir , microbiology and biotechnology , open reading frame , biology , chinese mitten crab , catalase , biochemistry , peptide sequence , untranslated region , complementary dna , amino acid , antioxidant , messenger rna , gene , zoology
Summary Catalase ( CAT ) is an antioxidant enzyme and plays a significant role in the protection against oxidative stress by reducing hydrogen peroxide. The CAT c DNA of E riocheir sinensis ( E s CAT ) was cloned via RACE technique. The complete sequence of E s CAT c DNA consisted of a 5′ untranslated regions ( UTR ) of 224 bp, a 3′ UTR of 1287 bp with a poly ( A ) tail and an open reading frame ( ORF ) of 1542 bp, which encoded a polypeptide of 513 amino acid residues with a calculated molecular mass of approximately 58.86 kDa and a theoretical isoelectric point of 6.880. The deduced amino acid sequence of E s CAT contained a highly conserved proximal active‐site signature motif ( 60 FDRERIPERVVHAKGAL 76 ) and a proximal heme–ligand signature motif ( 350 RLFSYNDTH 358 ) and exhibited high similarity with other reported CAT s. In the phylogenetic tree, E s CAT was clustered with the CAT s from S cylla serrata and P ortunus trituberculatus . The E s CAT transcripts were constitutively expressed in haepatopancreas, haemocytes, gill, gonad, muscle and heart, with highest expression level in haepatopancreas. The relative expression level of E s CAT m RNA in haemocytes was continuously up‐regulated and reached the peak level at 48 h post‐ V ibrio anguillarum challenge. The purified recombinant E s CAT protein displayed antioxidant activity against hydrogen peroxide with high thermal stability and broad spectrum of p H values. All these results demonstrated that E s CAT was an efficient antioxidant enzyme and potentially involved in the regulation of redox and innate immune response of crabs.