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Soluble Expression of Antimicrobial Peptide BSN-37 from Escherichia coli by SUMO Fusion Technology

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Abstract

Antimicrobial peptides (AMPs) are a kind of small molecular peptide that an organism produces to resist the invasion of foreign microorganisms. AMP BSN-37 is a bovine AMP that exhibits high antibacterial activity. In this paper, the optimized gene AMP BSN-37 was cloned into pCold-SUMO for fusion expression by recombinant DNA technology. The gene sequence of AMP BSN-37 was obtained by codons reverse translation, and the codons were optimized according to the codons preference of Escherichia coli (E. coli). The recombinant plasmid was constructed and identified by PCR, enzyme digestion and sequencing. Then the recombinant plasmid was transformed into BL21 E. coli to induce expression, and the IPTG concentration and time were optimized. The expressed soluble fusion protein SUMO-BSN-37 was purified by chromatography and then cleaved by SUMO proteases to release BSN-37. SDS-PAGE electrophoresis and Western blotting were used for identification. The recombinant plasmid pCold-SUMO-BSN-37 was obtained, and the fusion AMP BSN-37 was preliminarily expressed in BL21. After optimization, the optimal expression condition was 37 ℃ with 0.4 µM IPTG and 6 h incubation. Under optimal conditions, a large amount of fusion AMP BSN-37 was obtained by purification. Western blotting showed that the fusion peptide was successfully expressed and had good activity. The expressed BSN-37 showed antimicrobial activity similar to that of synthesized BSN-37. In this study, soluble expression products of AMP BSN-37 were obtained, and the problem regarding the limited source of AMP BSN-37 could be effectively solved, laying a foundation for further research on AMP BSN-37.

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Data Availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Change history

  • 15 April 2024

    ‘of’ word added in affiliation.

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Funding

This research was funded by the National Natural Science Foundation of China (32273042), the Science and Technology Innovation Team Support Program of the University of Henan Province (20IRTSTHN025), Postdoctoral Research Sponsorship in Henan Province (2015105), and Scientific and Technological Breakthrough Foundation of Henan Province (232102110086).

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Authors and Affiliations

  1. Postdoctoral Research Station, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450002, China

    Yanzhao Xu & Gaiping Zhang

  2. Postdoctoral Research Base, Henan Institute of Science and Technology, Xinxiang, 453003, China

    Yanzhao Xu, Jianhe Hu & Gaiping Zhang

  3. College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, 453003, China

    Yanzhao Xu, Mengmeng Dong, Qing Wang, Yawei Sun, Bolin Hang, Huihui Zhang & Jianhe Hu

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  1. Yanzhao Xu
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  2. Mengmeng Dong
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  3. Qing Wang
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  4. Yawei Sun
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  5. Bolin Hang
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  6. Huihui Zhang
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  7. Jianhe Hu
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Contributions

Data curation, Y.X. and M.D; Funding acquisition, Y.X. and J.H.; Methodology, Q.W., B.H. and Y.S.; Resources, H.Z. and Y.X.; Supervision, G.P.

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Correspondence to Yanzhao Xu or Gaiping Zhang.

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Xu, Y., Dong, M., Wang, Q. et al. Soluble Expression of Antimicrobial Peptide BSN-37 from Escherichia coli by SUMO Fusion Technology. Protein J 42, 563–574 (2023). https://doi.org/10.1007/s10930-023-10144-2

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