In vitro and in vivo characterization of a new recombinant antimicrobial peptide, MP1102, against methicillin-resistant Staphylococcus aureus
In vitro and in vivo characterization of a new recombinant antimicrobial peptide, MP1102, against methicillin-resistant Staphylococcus aureus
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DOI：
10.1007/s00253-015-6394-7
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发表时间：
2015-01
影响因子：
5
通讯作者：
Yong Zhang;D. Teng;Xiumin Wang;R. Mao;Xintao Cao;Xiaoyuan Hu;Lifen Zong;Jianhua Wang
中科院分区：
文献类型：
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作者：
Yong Zhang;D. Teng;Xiumin Wang;R. Mao;Xintao Cao;Xiaoyuan Hu;Lifen Zong;Jianhua Wang
目前，迫切需要更多的抗菌药物候选药物来应对病原微生物耐药性的上升。设计、鉴定了一种新的抗菌肽MP 1102，并在毕赤酵母中进行了高效表达。培养物中的总分泌蛋白达到695 mg/l，并且重组MP 1102（rMP 1102）的浓度为292 mg/l。通过一步阳离子交换色谱从发酵上清液中纯化rMP 1102，得到197.1 mg/l的产量，纯度为96.4%。rMP 1102对革兰氏阳性菌有较强的抗菌活性，对4种金黄色葡萄球菌（S.对20株MRSA临床分离株（cMRSA）的MIC为0.04 ~ 0.23 μM，比rNZ 2114（MIC为0.11 ~ 0.90 μM）强。rMP 1102显示杀死超过99.9%的测试S。在1、2、4倍MIC浓度下处理6 h内，金黄色葡萄球菌细胞死亡率达90%以上。在小鼠大腿感染模型中，浓度为5、10和20 mg/kg时，在12 h内对金黄色葡萄球菌细胞产生毒性。MRSA对MP 1102的敏感性高于其亲本肽NZ 2114，这一初步药效学分析表明这两种分子的杀伤机制可能存在差异。rMP 1102在128 μg/ml时引起小于0.05%的溶血活性，并在20至80 °C下表现出良好的热稳定性，在pH 8.0下观察到其最高活性。结果表明，该酵母表达系统具有规模化生产的可行性，rMP 1102具有较强的抗S.金霉素NZ 2114通过不同的作用机制对S.金黄色葡萄球菌，尤其是MRSA感染。
Currently, more antimicrobial drug candidates are urgently needed to combat the rise in drug-resistance among pathogenic microbes. A new antimicrobial peptide, MP1102, a variant of NZ2114, was designed, evaluated, and overexpressed inPichia pastoris. The total secreted protein in cultures reached 695 mg/l, and the concentration of the recombinant MP1102 (rMP1102) was 292 mg/l. rMP1102 was purified from the fermentation supernatant by one-step cation exchange chromatography to obtain a yield of 197.1 mg/l with 96.4 % purity. rMP1102 exhibited potent activity against Gram-positive bacteria, and its minimum inhibitory concentrations (MICs) for fourStaphyloccocus aureus(S. aureus) strains ranged from 0.028 to 0.11 μM, and it had stronger activity (MIC = 0.04 to 0.23 μM) to 20 clinical isolates of MRSA (cMRSA) than rNZ2114 (MIC = 0.11 to 0.90 μM). rMP1102 was shown to kill over 99.9 % of testedS. aureuscells within 6 h when treated at one, two, and four times its MIC and over 90 % ofS. aureuscells within 12 h at concentrations of 5, 10, and 20 mg/kg in a mouse thigh infection model. The higher sensitivity of MRSA to MP1102 than to its parental peptide, NZ2114, indicated by this initial pharmacodynamic analysis suggests a possible difference in the killing mechanism of these two molecules. rMP1102 caused less than 0.05 % hemolytic activity at 128 μg/ml and exhibited good thermostability from 20 to 80 °C, with its highest activity being observed at pH 8.0. These results suggest that this yeast expression system is feasible for large-scale production, and rMP1102 exerted stronger activity againstS. aureusthan NZ2114 via a different mechanism and exhibited potential as a new antimicrobial agent forS. aureus, especially MRSA infections.