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In vitro and in vivo characterization of a new recombinant antimicrobial peptide, MP1102, against methicillin-resistant Staphylococcus aureus

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Abstract

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 in Pichia 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 four Staphyloccocus 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 tested S. aureus cells within 6 h when treated at one, two, and four times its MIC and over 90 % of S. aureus cells 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 against S. aureus than NZ2114 via a different mechanism and exhibited potential as a new antimicrobial agent for S. aureus, especially MRSA infections.

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (No. 31372346 and No. 31302004), the Project of the National Support Program for Science and Technology in China (No. 2013BAD10B02 and No. 2011BAD26B02), the Special Fund for Agro-scientific Research in the Public Interest in China (No. 201403047), and the AMP Direction of National Innovation Program of Agricultural Science and Technology in CAAS (2013–2017).

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    Authors and Affiliations

    1. Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China

      Yong Zhang, Da Teng, Xiumin Wang, Ruoyu Mao, Xintao Cao, Xiaoyuan Hu, Lifen Zong & Jianhua Wang

    2. Gene Engineering Laboratory, Feed Research Institute, Chinese Academy of Agricultural Sciences, 12 Zhongguancun Nandajie St., Haidian District, 100081, Beijing, China

      Yong Zhang, Da Teng, Xiumin Wang, Ruoyu Mao, Xintao Cao, Xiaoyuan Hu, Lifen Zong & Jianhua Wang

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    1. Yong Zhang
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    Correspondence to Jianhua Wang.

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    Yong Zhang and Da Teng contributed equally to this work.

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    Zhang, Y., Teng, D., Wang, X. et al. In vitro and in vivo characterization of a new recombinant antimicrobial peptide, MP1102, against methicillin-resistant Staphylococcus aureus . Appl Microbiol Biotechnol 99, 6255–6266 (2015). https://doi.org/10.1007/s00253-015-6394-7

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