Abstract
Objective
We aimed to clone and express the human Cu, Zn superoxide dismutase (hSOD1) in Bacillus subtilis 1012. Also, we investigated the expression level of hSOD1 under different induction conditions.
Result
As an essential member of the antioxidant defense system in vivo, hSOD1 has become a therapeutic agent against host diseases, such as oxygen toxicity, acute inflammation, and radiation injury. The recombinant hSOD1 was successfully secreted extracellularly into B. subtilis 1012. The expression conditions were optimized, including inoculum size, different media, temperatures, and inducer concentrations. Finally, the highest level of hSOD1 was produced as a soluble form in Super rich medium by 2% inoculum with 0.2 mM of IPTG at 37 °C after the induction for 24 h. Besides, 20 g/L of lactose also displayed the same inductive effect on hSOD1 expression as that of IPTG (0.2 mM). Finally, the specific activity of purified hSOD1 was determined to be 1625 U/mg in the presence of 800 μM of Cu2+ and 20 μM of Zn2+.
Conclusions
We propose that the B. subtilis 1012-hSOD1 strain system has great potential in future industrial applications.
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Funding
This work was supported by Educational Commission of Hubei Province of China (No. Q20212012), Department of Science and Technology of Hubei Province (No.2021CFB253), Key Research and Development Plan of Ningxia Autonomous Region (No. 2021ZDYF0552), and Project of Excellent Young and Middle-aged Scientific and Technological Innovation Team in Colleges and Universities of Hubei Province (No. T2020013).
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Hongtao Liu and Zhigang Zhang designed the study. Mingzhu Yin, Nian Wang, Qiqi Wang, Hui Xia, Xue Cheng, and Haiming Hu were responsible for the acquisition of data. Mingzhu Yin and Nian Wang interpreted the experimental data. Mingzhu Yin and Hongtao Liu were the major contributors in drafting and revising the manuscript. All authors have read and approved the final manuscript.
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Yin, M., Wang, N., Wang, Q. et al. Cloning and expression of recombinant human superoxide dismutase 1 (hSOD1) in Bacillus subtilis 1012. Biotechnol Lett 45, 125–135 (2023). https://doi.org/10.1007/s10529-022-03319-x
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DOI: https://doi.org/10.1007/s10529-022-03319-x