A superoxide dismutase from salt lake, recombinant protein and preparation method and application thereof
By screening recombinant protein D10-2 from salt lake soil and expressing it in Escherichia coli, the problem of insufficient stability of natural superoxide dismutase under extreme environments has been solved, achieving high stability at high temperatures and over a wide pH range, thus expanding its application in antioxidant drugs and cosmetics.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
- Filing Date
- 2026-03-05
- Publication Date
- 2026-06-16
AI Technical Summary
Existing natural superoxide dismutases lack stability under industrial conditions such as high temperature, extreme pH, and high salt, which limits their large-scale application.
The mesothermal superoxide dismutase D10-2 gene was screened from the soil of Dabancheng Salt Lake in Xinjiang. It was expressed and purified in Escherichia coli DH5α using recombinant protein technology. The obtained recombinant protein D10-2 maintained high stability in the range of 30-50℃ and pH 5-10.
Recombinant protein D10-2 retains 100% residual activity after incubation at 45℃ for 14 hours, and retains more than 90% residual activity after incubation at pH 5.0-10.0 for 12 hours, making it suitable for anti-oxidative disease drugs and heat-resistant cosmetics.
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Figure CN122214291A_ABST
Abstract
Claims
1. A mesophilic superoxide dismutase, characterized in that, The mesothermal superoxide dismutase is named D10-2, and the DNA sequence of D10-2 is shown in SEQ ID NO:
1.
2. The mesophilic superoxide dismutase according to claim 1, characterized in that, The D10-2 was screened from salt lake soil.
3. The mesophilic superoxide dismutase according to claim 1, characterized in that, The activity stability temperature of D10-2 is 30-50℃, and the activity stability is maintained when the pH value is in the range of 5-10.
4. A recombinant protein prepared using a mesophilic superoxide dismutase according to any one of claims 1-3, wherein the DNA sequence of the recombinant protein is shown in SEQ ID NO:
2.
5. A method for preparing a recombinant superoxide dismutase protein at medium temperature according to claim 4, characterized in that, Includes the following steps: Step 1: Based on functional prediction, the sequence D10-2 of a superoxide dismutase gene, as shown in SEQ ID NO:1, was isolated from the metagenomic database. Step 2: The DNA sequence shown in SEQ ID NO:1 is seamlessly cloned into the expression vector to obtain the recombinant plasmid; Step 3: Transform the recombinant plasmid into recipient cells to obtain recombinant bacteria; Step 4: Extract and purify the recombinant protein from the recombinant bacteria.
6. The preparation method according to claim 5, characterized in that, The vector is a constitutive expression plasmid vector pSHY211, and the recombinant plasmid obtained is pSHY211-d10-2.
7. The preparation method according to claim 5, characterized in that, The recipient cells were Escherichia coli DH5α, and the transformation method was electroporation transformation.
8. The use of the mesophilic superoxide dismutase according to any one of claims 1-3 in the preparation of drugs for treating antioxidant diseases.
9. The application according to claim 8, characterized in that, The mesophilic superoxide dismutase is used as an effective component in the preparation of antioxidant disease drugs.
10. The application of the mesophilic superoxide dismutase according to any one of claims 1-3 in the preparation of heat-resistant cosmetics.