摘要
抗菌肽因其具有广谱抑菌活性及不易引起微生物耐药性的特点,具有成为新型抗菌药物的潜力。为研究植物源抗菌肽MaSAMP在毕赤酵母中的表达及活性,利用PCR扩增MaSAMP基因片段,并与组成型表达载体pGAPZαC连接,电转至毕赤酵母GS115感受态细胞中,构建高效表达MaSAMP的毕赤酵母工程菌株,并进一步探讨重组抗菌肽MaSAMP对不同病原菌的抑菌活性。结果表明,根据毕赤酵母密码子偏好性优化植物源抗菌肽MaSAMP的密码子,成功将其与表达载体pGAPZαC连接,并电转至毕赤酵母GS115,实现了抗菌肽MaSAMP在毕赤酵母中的高效表达。抗菌肽MaSAMP对青枯雷尔氏菌、胡萝卜软腐果胶杆菌、黄单胞杆菌等植物病原菌具有较强的抑菌效果。通过优化培养条件发现,抗菌肽MaSAMP在培养基初始pH 6.5、培养温度28℃、转速220 r·min^(-1)、摇瓶72 h时表现出最佳的抑菌活性。抗菌肽MaSAMP在不同温度、pH、紫外照射时间及金属离子处理下的稳定性存在差异。以上研究结果为抗菌肽MaSAMP的生产应用提供了理论基础及技术参考,并为植物细菌性病害防治提供了新的思路。
Antimicrobial peptides(AMPs)have the potential to become novel antimicrobial agents due to their broadspectrum antibacterial activity and low propensity for inducing microbial resistance.To investigate the expression and activity of the plant-derived antimicrobial peptide MaSAMP in Pichia pastoris,and the MaSAMP gene fragment was amplified by PCR and ligated with the constitutive expression vector pGAPZαC,and then electrotransferred into the cells of Pichia pastoris GS115 to construct the Pichia pastoris engineered strain with high efficiency in expressing MaSAMP.The inhibitory effect of the plant-derived antimicrobial peptide MaSAMP against various plant pathogenic bacteria were evaluated.The results showed that,by optimizing the codons of the plant-derived antimicrobial peptide MaSAMP according to the codon preferences of Pichia pastoris,the peptide was successfully ligated into the expression vector pGAPZαC and transformed into Pichia pastoris GS115,achieving high-level expression of MaSAMP in the yeast.The antimicrobial peptide MaSAMP exhibited strong inhibitory effects against plant pathogenic bacteria such as Ralstonia solanacearum,Pectobacterium carotovorum and Xanthomonas campestris.Furthermore,optimization of culture conditions revealed that the best antibacterial activity of MaSAMP was observed with initial pH 6.5,culture temperature of 28℃,shaking speed of 220 r·min^(-1) and culture time of 72 h.The stability of MaSAMP varied under different temperatures,pH,UV exposure times and metal ion treatments.Above results provided theoretical basis and technical reference for the production and application of antimicrobial peptide MaSAMP,and offered new approach for the prevention and control of bacterial diseases in plants.
作者
查曼
王常高
刘晓艳
李斐
朱镭
邱一敏
陈凌
周荣华
闵勇
ZHA Man;WANG Changgao;LIU Xiaoyan;LI Fei;ZHU Lei;QIU Yimin;CHEN Ling;ZHOU Ronghua;MIN Yong(Hubei Biopesticide Engineering Research Center,Wuhan 430064,China;School of Life and Health Science,Hubei University of Technology,Wuhan 430068,China;Institute of Plant Protection and Soil Fertilizer,Hubei Academy of Agricultural Sciences,Wuhan 430064,China)
出处
《中国农业科技导报(中英文)》
北大核心
2026年第4期151-160,共10页
Journal of Agricultural Science and Technology
基金
武汉市知识创新专项曙光计划项目(2023020201020433)
湖北省技术创新计划项目(2024BBB064)
湖北省现代农业产业技术体系项目(2023HBSTX4-07)。
关键词
抗菌肽
毕赤酵母
发酵优化
稳定性
植物病原菌
antimicrobial peptides
Pichia pastoris
fermentation optimization
stability
plant pathogens
