数字农科院2.0

Design and recombination expression of a novel plectasin-derived peptide MP1106 and its properties against Staphylococcus aureus

文献类型： 外文期刊

作者： Cao, Xintao;Zhang, Yong;Mao, Ruoyu;Teng, Da;Wang, Xiumin;Wang, Jianhua;Cao, Xintao;Zhang, Yong;Mao, Ruoyu;Teng, Da;Wang, Xiumin;Wang, Jianhua

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关键词： MP1106;Antimicrobial peptide;Pichia pastoris;Stability;Staphylococcus aureus

期刊名称： APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

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收录情况： SCI

摘要： A novel antimicrobial peptide MP1106 was designed based on the parental peptide plectasin with four mutational sites and a high level of expression in Pichia pastoris X-33 via the pPICZ alpha A plasmid was achieved. The concentration of total secreted protein in the fermented supernatant was 2.134 g/l (29 A degrees C), and the concentration of recombinant MP1106 (rMP1106) reached 1,808 mg/l after a 120-h induction in a 5-l fermentor. The rMP1106 was purified using a cation-exchange column, and the yield was 831 mg/l with 94.68 % purity. The sample exhibited a narrow spectrum against some Gram-positive bacteria and strong antimicrobial activity against Staphylococcus aureus at low minimal inhibitory concentrations (MICs) of 0.014, 1.8, 0.45, and 0.91 mu M to S. aureus strains ATCC 25923, 29213, 6538, and 43300, respectively. Meanwhile, rMP1106 showed potent activity (0.03-1.8 mu M) against 20 clinical isolates of methicillin-resistant S. aureus (MRSA). In addition, rMP1106 exhibited a broad range of thermostability from 20 to 100 A degrees C. The higher antimicrobial activity of rMP1106 was maintained in neutral and alkaline environments (pH 6, 8, and 10), and its activity was slightly reduced in acidic environments (pH 2 and 4). The rMP1106 was resistant to the digestion of pepsin, snailase, and proteinase K and was sensitive to trypsin. It exhibited hemolytic activity of only 1.16 % at a concentration of 512 mu g/ml and remained stable in human serum at 37 A degrees C for 24 h. Furthermore, the activity of rMP1106 was minorly affected by 10 mM dithiothreitol and 20 % dimethylsulfoxide. Our results indicate that MP1106 can be produced on a large scale and has potential as a therapeutic drug against S. aureus.

分类号： Q939.9`Q81

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