Department of Gastrointestinal Surgery, China-Japan Union Hospital, Jilin University, Changchun 130000, China.
Center of Chinese Medicine and Bio-Engineering Research, Changchun University of Chinese Medicine, Changchun 130117, China.
Molecules. 2014 Jun 16;19(6):8112-23. doi: 10.3390/molecules19068112.
Superoxide dismutase (SOD) has a strong antioxidant effect, but the traditional SOD extraction method is not the most efficient method of SOD amplification. In this study, we report the cloning of the Cu/ZnSOD gene from Panax ginseng into a temperature-regulated expression plasmid, pBV220. Cu/ZnSOD inclusion bodies were expressed in E. coli at a high level. Then, the inclusion bodies were purified by ion-exchange chromatography and molecular sieve chromatography. Finally, we obtained stable SOD in the bacterial broth, with a protein content of 965 mg/L and enzyme specific activity of 9389.96 U/mg. These results provide a foundation for future studies on the antioxidant mechanisms of ginseng and the development and application of ginseng Cu/ZnSOD.
超氧化物歧化酶(SOD)具有很强的抗氧化作用,但传统的 SOD 提取方法不是 SOD 扩增的最有效方法。在这项研究中,我们报告了从人参中克隆铜/锌 SOD 基因到温度调节表达质粒 pBV220 的情况。Cu/ZnSOD 包涵体在大肠杆菌中高水平表达。然后,通过离子交换层析和分子筛层析对包涵体进行纯化。最后,我们从细菌培养液中获得了稳定的 SOD,其蛋白含量为 965mg/L,酶比活为 9389.96U/mg。这些结果为今后研究人参的抗氧化机制以及人参 Cu/ZnSOD 的开发和应用奠定了基础。