Abstract
As the most important member of antioxidant defense system, human Cu,Zn superoxide dismutase (hCu,Zn-SOD) protects cells against the free radicals produced by aerobic metabolism. hCu,Zn-SOD has been widely used in food, cosmetic and medicine industry due to its health benefits and therapeutic potentials. However, a more extensive application of hCu,Zn-SOD is limited by the challenge of expensive and low production of high-activity hCu,Zn-SOD in large scale. In this study, the codon-optimized hCu,Zn-SOD gene was synthesized, cloned into pET-28a( +) and transformed into Escherichia coli BL21(DE3). After induction with IPTG or lactose, hCu,Zn-SOD was highly expressed as soluble form in LB medium with 800 μM Cu2+ and 20 μM Zn2+ at 25 °C. The recombinant hCu,Zn-SOD was efficiently purified by nickel affinity chromatography. Through optimization of fed-batch fermentation conditions, 342 mg purified hCu,Zn-SOD was obtained from 1 L cultures fermented in a 3-L bioreactor. Furthermore, the recombinant hCu,Zn-SOD retained the enzymatic specific activity of 46,541 U/mg. This study has opened up an effective avenue for industrial production of hCu,Zn-SOD through microbial fermentation in the future.
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This work is supported by grants from the Drug Innovation Major Project (2018ZX09711001-006) and CAMS Innovation Fund for Medical Sciences (CIFMS) (2019-I2M-005).
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YJL and ZP designed this project. LXL and YJL performed the experiments. All the authors analyzed the data. YJL wrote the manuscript. All authors read and approved the final manuscript.
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Yang, JL., Li, XL., Jiang, FL. et al. High-level soluble expression of human Cu,Zn superoxide dismutase with high activity in Escherichia coli. World J Microbiol Biotechnol 36, 106 (2020). https://doi.org/10.1007/s11274-020-02883-6
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DOI: https://doi.org/10.1007/s11274-020-02883-6