Effect of cell-permeable grouper Manganese Superoxide Dismutase on environmental stress in fish

Nitrite levels are generally high in high-density aquaculture. Nitrite is a potential stress-inducing factor and can cause oxidative stress because excessive reactive oxygen species (ROS) formation through nitrite induction cannot be scavenged by the endogenous antioxidant system, thus leading to cell damage or death. Manganese Superoxide Dismutase (MnSOD) is a highly efficient endogenous ROS scavenger that quenches mitochondrial ROS and protective against oxidative stress. To enhance the efficiency of MnSOD in removing ROS and reducing oxidative caused by nitrite, in this study, we cloned grouper MnSOD (gMnSOD) fused with a cell-penetrating peptide, TAT, to construct a TAT-gMnSOD fusion protein and assessed its potential to eliminate excess ROS induced by high nitrite concentrations and enhance the resistance of zebrafish to environmental stressors. Our results revealed that TAT-gMnSOD penetrated the grouper fin (GF-1) cells, scavenged nitrite-induced intracellular ROS, and enhanced cell viability on NaNO₂ treatment. Furthermore, pretreatment of zebrafish with TAT-gMnSOD fusion protein reduced the MDA content and increased the survival rate. In addition, the TAT-gMnSOD fusion protein reduced 2-phenoxyethanol toxicity and attenuated excessive anesthesia among zebrafish. In conlusion, our cell-permeable TAT-gMnSOD fusion protein effectively counters oxidative stress, prevents environmental stress-induced damage, and increases aquaculture benefits.