Manganese superoxide dismutase (MnSOD) is an antioxidant that exists in mitochondria and can effectively remove superoxide anions in mitochondria. In a dark, high-pressure, and low-temperature deep-sea environment, MnSOD is essential for the survival of sea cucumbers. Six MnSODs were identified from the transcriptomes of deep and shallow-sea sea cucumbers. To explore their environmental adaptation mechanism, we conducted environmental selection pressure analysis through the branching site model of PAML software. We obtained night positive selection sites, and two of them were significant (97F -> H, 134K -> V): 97F -> H located in a highly conservative characteristic sequence, and its polarity change might have a great impact on the function of MnSOD; 134K -> V had a change in piezophilic ability, which might help MnSOD adapt to the environment of high hydrostatic pressure in the deepsea. To further study the effect of these two positive selection sites on MnSOD, we predicted the point mutations of F97H and K134V on shallow-sea sea cucumber by using MAESTROweb and PyMOL. Results show that 97F -> H, 134K -> V might improve MnSOD's efficiency of scavenging superoxide anion and its ability to resist high hydrostatic pressure by moderately reducing its stability. The above results indicated that MnSODs of deep-sea sea cucumber adapted to deep-sea environments through their amino acid changes in polarity, piezophilic behavior, and local stability. This study revealed the correlation between MnSOD and extreme environment, and will help improve our understanding of the organism's adaptation mechanisms in deep sea.