食品与机械2025，Vol.41Issue(8)：39-45,7.DOI:10.13652/j.spjx.1003.5788.2025.60053
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
Prokaryotic expression of recombinant Plectasin optimization of high-density fermentation conditions and antibacterial activity
摘要
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 pET32a-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 BL21(DE3)/pLysS/pET32a-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 fermentation conditions improved the soluble expression yield of recombinant Plectasin,and the fermentation product exhibited antibacterial activity against S.aureus.关键词
高密度发酵/可溶性表达/Plectasin/分批补料培养/抑菌活性Key words
high-density fermentation/soluble expression/Plectasin/fed batch culture/antibacterial activity引用本文复制引用
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):39-45,7.基金项目
安徽省教育厅高校自然科学重点研究项目(编号:KJ2021A1386) （编号:KJ2021A1386）
安徽省高校青年骨干人才国内访学研修项目(编号:gxgnfx2021199) （编号:gxgnfx2021199）
合肥职业技术学院产学研团队项目(编号:2024CXYTD02) （编号:2024CXYTD02）
合肥职业技术学院生物医药协同创新平台项目(编号:2024PTKJA01) （编号:2024PTKJA01）
