Design, expression, and characterization of a novel targeted plectasin against methicillin-resistant Staphylococcus aureus

文献类型： 外文期刊

第一作者： Mao, Ruoyu

作者： Mao, Ruoyu;Teng, Da;Wang, Xiumin;Xi, Di;Zhang, Yong;Hu, Xiaoyuan;Yang, Yalin;Wang, Jianhua;Mao, Ruoyu;Teng, Da;Wang, Xiumin;Xi, Di;Zhang, Yong;Hu, Xiaoyuan;Wang, Jianhua

作者机构：

关键词： Agplectasin;STAMP;Plectasin.MRSA;Pichia pastoris

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

ISSN：

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页码：

收录情况： SCI

摘要： A novel specifically targeted antimicrobial peptide (STAMP) that was especially effective against methicillin-resistant Staphylococcus aureus (MRSA) was designed by fusing the AgrD1 pheromone to the Nterminal end of plectasin. This STAMP was named Agplectasin, and its gene was synthesized and expressed in Pichia pastoris X-33 via pPICZαA. The highest amount of total secreted protein reached 1,285.5 mg/l at 108 h during the 120-h induction. The recombinant Agplectasin (rAgP) was purified by cation exchange chromatography and hydrophobic exchange chromatography; its yield reached 150 mg/l with 94 % purity. The rAgP exhibited strong bactericidal activity against S. aureus but not Staphylococcus epidermidis or other types of tested bacteria. A bactericidal kinetics assay showed that the rAgP killed over 99.9 % of tested S. aureus (ATCC 25923 and ATCC 43300) in both Mueller-Hinton medium and human blood within 10 h when treated with 4× minimal inhibitory concentration. The rAgP caused only approximately 1 % hemolysis of human blood cells, even when the concentration reached 512 μg/ml, making it potentially feasible as a clinical injection agent. In addition, it maintained a high activity over a wide range of pH values (2.0-10.0) and demonstrated a high thermal stability at 100 °C for 1 h. These results suggested that this STAMP has the potential to eliminate MRSA strains without disrupting the normal flora.

分类号： Q939.9

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