Cu/Zn superoxide dismutases (SODs) are antioxidative metalloenzymes that exist ubiquitously in different species and are distributed widely in various tissues and cell types. In this study, the distribution and biological function of the Cu/Zn superoxide dismutase in Apostichopus japonicus (AjSOD1) is first characterized. The AjSOD1 cDNA is 1219 bp in length and contains an open reading frame (ORF) of 459 bp that encodes a protein of 152 amino acids with a deduced molecular weight of 15.47 kDa and a predicted isoelectric point of 5.65. The Cu2+/Zn2+ binding domain and conserved residues were found in the AjSOD1 amino acid sequence. A quantitative reverse transcriptase real-time PCR (qRT-PCR) assay was developed to assess the expression of AjSOD1 in different tissues. Spatial distribution analysis showed that AjSOD1 was constitutively expressed in all tested tissues, with strong expression in the intestine and weak expression in the respiratory tree. mRNA Expression of AjSOD1 was significantly upregulated when challenged with the pathogen Vibrio splendidus. Functional investigation revealed that recombinant AjSOD1 displayed good antioxidant activity. More importantly, the addition of AjSOD1 resulted in a significant decrease in coelomocyte apoptosis by LPS/H2O2 challenge in vitro. The results indicate that sea cucumber SOD1 may play critical roles not only in the defense against oxidative stress but also in the innate immune defense against bacterial infections.
Keywords: Antioxidative activity; Apostichopus japonicus; Cu/Zn-SOD; Recombinant protein; Spatial expression.
Copyright © 2019 Elsevier Ltd. All rights reserved.