Abstract
[Objective] To construct a recombinant strain expressing the antimicrobial peptide Plectasin, optimize the high-density fermentation conditions of the recombinant strain, and evaluate the antibacterial activity of the purified recombinant peptide produced under optimized conditions. [Methods] To improve soluble expression of the target protein, the plectasin gene was synthesized and optimized based on codon preference of the prokaryotic expression system with signal peptide deletion. The optimized gene was cloned into the pET32a(+) expression vector to construct the recombinant plasmid pET 32a-plectasin, which was transformed into Escherichia coli BL21(DE3)/pLysS competent cells. Single-factor and orthogonal experiments were designed to optimize the parameters of 20 L high-density fermentation for the recombinant strain. The recombinant Plectasin produced under optimized conditions was purified and its antibacterial activity was analyzed. [Results] The E.coli BL 21(DE3)/pLysS/pET 32a-plectasin expression strain was successfully constructed. The optimized 20 L high-density fermentation conditions were induction temperature of 30℃, inoculum size of 2%, and fermentation duration of 15 h. After purification, the recombinant protein concentration reached (0.62±0.02) g/L. The minimum inhibitory concentration (MIC) of the expressed product against Staphylococcus aureus ATCC 25923 was 32 μg/mL. [Conclusion] Optimization of prokaryotic high-density 引用格式：苏琰，李融. 重组抗菌肽 Plectasin 的原核表达高密度发酵条件优化及抑菌活性 [J]. 食品与机械，2025，41(8):39-45. Citation:SU Yan, LI Rong. Prokaryotic expression of recombinant Plectasin optimization of high-density fermentation conditions and antibacterial activity [J]. Food & Machinery, 2025, 41(8):39-45. fermentation conditions improved the soluble expression yield of recombinant Plectasin, and the fermentation product exhibited antibacterial activity against S.aureus.
Publication Date
9-25-2025
First Page
39
Last Page
45
DOI
10.13652/j.spjx.1003.5788.2025.60053
Recommended Citation
Yan, SU and Rong, LI
(2025)
"Prokaryotic expression of recombinant Plectasin optimization of high-density fermentation conditions and antibacterial activity,"
Food and Machinery: Vol. 41:
Iss.
8, Article 5.
DOI: 10.13652/j.spjx.1003.5788.2025.60053
Available at:
https://www.ifoodmm.cn/journal/vol41/iss8/5
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