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Mar Drugs
2019 Feb 01;172:. doi: 10.3390/md17020084.
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Characteristics of a Novel Manganese Superoxide Dismutase of a Hadal Sea Cucumber (Paelopatides sp.) from the Mariana Trench.
Li Y
,
Kong X
,
Zhang H
.
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A novel, cold-adapted, and acid-base stable manganese superoxide dismutase (Ps-Mn-SOD) was cloned from hadal sea cucumber Paelopatides sp. The dimeric recombinant enzyme exhibited approximately 60 kDa in molecular weight, expressed activity from 0 °C to 70 °C with an optimal temperature of 0 °C, and resisted wide pH values from 2.2⁻13.0 with optimal activity (> 70%) at pH 5.0⁻12.0. The Km and Vmax of Ps-Mn-SOD were 0.0329 ± 0.0040 mM and 9112 ± 248 U/mg, respectively. At tested conditions, Ps-Mn-SOD was relatively stable in divalent metal ion and other chemicals, such as β-mercaptoethanol, dithiothreitol, Tween 20, Triton X-100, and Chaps. Furthermore, the enzyme showed striking stability in 5 M urea or 4 M guanidine hydrochloride, resisted digestion by proteases, and tolerated a high hydrostatic pressure of 100 MPa. The resistance of Ps-Mn-SOD against low temperature, extreme acidity and alkalinity, chemicals, proteases, and high pressure make it a potential candidate in biopharmaceutical and nutraceutical fields.
Figure 1. Nucleotide and corresponding amino acid sequences of Ps-Mn-SOD. The signal peptide is drawn with a red line. The signature sequence DVWEHAYY is underlined with dotted line. N- and C-terminal domains are marked with purple and green shades, respectively. Four conserved amino acid residues for manganese coordination are boxed. Asterisk points to the highly conserved Tyr-35 residue. Cylinders and arrows represent helices and strands, respectively.
Figure 2. Multiple alignment of Ps-Mn-SOD with other invertebrates. Mn-SOD signature sequence is boxed. Triangles point to the active sites for manganese coordination. Asterisk points to the highly conserved Tyr-35 residue.
Figure 3. Neighbor-joining phylogenetic tree of SODs based on amino acid sequence homology. Bootstrap values below 50 are cut off. Ps-Mn-SOD is displayed in bold.
Figure 4. Effects of temperature (A), pH (B), urea and guanidine hydrochloride (C), and high hydrostatic pressure (D) on Ps-Mn-SOD. Ps-SOD and Be-SOD represent SOD from Paelopatides sp. and bovine erythrocytes, respectively.
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