Molecular cloning and functional characterization of a Cu/Zn superoxide dismutase from jellyfish Cyanea capillata
Molecular cloning and functional characterization of a Cu/Zn superoxide dismutase from jellyfish Cyanea capillata
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Cyanea capillata 水母铜/锌超氧化物歧化酶的分子克隆和功能表征
DOI：
10.1016/j.ijbiomac.2019.12.071
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Zhang Liming
中科院分区：
文献类型：
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作者：
Wang Bo;Liu Guoyan;Wang Chao;Ruan Zengliang;Wang Qianqian;Wang Beilei;Qiu Leilei;Zou Shuaijun;Zhang Xiping;Zhang Liming
我们从水母（cyanea capillata）触须组织的cDNA文库中鉴定并鉴定了一种新的超氧化物歧化酶（SOD），命名为CcSOD1。CcSOD1全长cDNA序列由745个核苷酸组成，开放阅读框编码154个氨基酸的成熟蛋白，与典型的Cu/Zn-SODs具有相似的预测结构。将CcSOD1编码序列克隆到表达载体pET-24a中，在大肠杆菌rosetta (DE3) pLysS中成功表达。重组蛋白rCcSOD1经HisTrap高效螯合柱层析纯化，并进行生物学功能分析。结果表明，纯化后的rCcSOD1在pH为4.5 ~ 9、温度为10 ~ 70℃范围内均能抑制超氧阴离子，并保持活性。即使在70°C下加热60分钟，rCcSOD1仍保持100%的活性，表明相对较高的热稳定性。这些结果表明，CcSOD1蛋白可能在保护水母免受氧化损伤中发挥重要作用，可以作为抗氧化产物的新资源。
We identified and characterized a novel superoxide dismutase (SOD), designated as CcSOD1, from the cDNA library from the tentacle tissue of the jellyfishCyanea capillata. The full-length cDNA sequence of CcSOD1 consists of 745 nucleotides with an open reading frame encoding a mature protein of 154 amino acids, sharing a predicted structure similar to the typical Cu/Zn-SODs. The CcSOD1 coding sequence was cloned into the expression vector pET-24a and successfully expressed inEscherichia coliRosetta (DE3) pLysS. The recombinant protein rCcSOD1 was purified by HisTrap High Performance chelating column chromatography and analyzed for its biological function. Our results showed that the purified rCcSOD1 could inhibit superoxide anion and keep active in a pH interval of 4.5–9 and a temperature interval of 10–70°C. Even when heated at 70°C for 60 min, rCcSOD1 retained 100% activity, indicating a relatively high thermostability. These results suggest that CcSOD1 protein may play an important role in protecting jellyfish from oxidative damage and can serve as a new resource for antioxidant products.