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Maltose Induced Expression of Cecropin AD by SUMO Technology in Bacillus subtilis WB800N

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Abstract

Cecropin AD (CAD) is a hybrid peptide composed of 37 amino acids with the characters of strong antibacterial, antitumor properties and no hemolytic activity, which was regarded as a promising antibiotic candidate. Thus, a safe method to produce Cecropin AD is necessary to be found. In the study, Bacillus subtilis WB800N was employed as host strain. The CAD coding sequence fused with the signal peptide of SPsacB, the 6 × His gene and the gene of small ubiquitin-like modifier were cloned into the maltose-inducible vector pGJ148. Under the induction by 6% maltose, the recombinant fusion protein was successfully expressed and detected in culture substrate. An optimized amount (26.4 mg/L) of the recombinant CAD was purified of culture supernatant. After purification and digestion, the recombinant CAD was harvested about 4.5 mg/L with a purity of 93%. Recombinant CAD exhibited similar antimicrobial activity with synthetic CAD. This shows that the production of CAD in maltose-induced Bacillus subtilis expression system is a relatively safe method, which is vital for the application of CAD in animal husbandry production.

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Funding

This work was supported by the Grants form Natural Science Foundation of China (31672434, 31872368, 31472104), the China Agriculture Research System (CARS-35), and the Natural Science Foundation of Heilongjiang Province (TD2019C001).

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  1. Licong Zhang and Xiaodan Li contributed to the work equally.

Authors and Affiliations

  1. Institute of Animal Nutrition, Northeast Agricultural University, Harbin, People’s Republic of China

    Licong Zhang, Xiaodan Li, Na Zhan, Taotao Sun, Jianping Li & Anshan Shan

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  1. Licong Zhang
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  2. Xiaodan Li
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Correspondence to Jianping Li or Anshan Shan.

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Zhang, L., Li, X., Zhan, N. et al. Maltose Induced Expression of Cecropin AD by SUMO Technology in Bacillus subtilis WB800N. Protein J 39, 383–391 (2020). https://doi.org/10.1007/s10930-020-09908-x

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