Mechanism of antimicrobial peptide NP-6 from Sichuan pepper seeds against E. coli and effects of different environmental factors on its activity

  • Xiaoyan Hou
  • Chaoyang Feng
  • Shanshan Li
  • Qingying Luo
  • Guanghui Shen
  • Hejun Wu
  • Meiliang Li
  • Xingyan Liu
  • Anjun Chen
  • Meng Ye
  • Zhiqing ZhangEmail author
Biotechnologically relevant enzymes and proteins


A novel antimicrobial peptide named NP-6 was identified in our previous work. Here, the mechanisms of the peptide against Escherichia coli (E. coli) were further investigated, as well as the peptide’s resistance to temperature, pH, salinity, and enzymes. The transmission electron microscopy (TEM), confocal laser scanning microcopy (CLSM), and flow cytometric (FCM) analysis, combined with measurement of released K+, were performed to evaluate the effect of NP-6 E. coli cell membrane. The influence of NP-6 on bacterial DNA/RNA and enzyme was also investigated. The leakage of K+ demonstrated that NP-6 could increase the permeability of E. coli cell membrane. The ATP leakage, FCM, and CLSM assays suggested that NP-6 caused the disintegration of bacterial cell membrane. The TEM observation indicated that NP-6 could cause the formation of empty cells and debris. Besides, the DNA-binding assay indicated that NP-6 could bind with bacterial genomic DNA in a way that ethidium bromide (EB) did, and suppress the migration of DNA/RNA in gel retardation. Additionally, NP-6 could also affect the activity of β-galactosidase. Finally, the effect of different surroundings such as heating, pH, ions, and protease on the antimicrobial activity of NP-6 against E. coli was also investigated. Results showed that the peptide was heat stable in the range of 60~100 °C and performed well at pH 6.0~8.0. However, the antimicrobial activity of NP-6 decreased significantly in the presence of Mg2+/Ca2+, and after incubation with trypsin/proteinase K. The results will provide a theoretical support in the further application of NP-6.


Antimicrobial peptide Zanthoxylum bungeanum Maxim seeds Mechanisms Antimicrobial activity Surroundings effects 


Funding information

This study received financial assistance from the Scientific Fund of the Application Fundamental Project (2016-JY-0118) and Key Project of Research and Development Program (18ZDYF1175), Science and Technology Department of Sichuan Province.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals by any of the authors.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Xiaoyan Hou
    • 1
  • Chaoyang Feng
    • 1
  • Shanshan Li
    • 1
  • Qingying Luo
    • 1
  • Guanghui Shen
    • 1
  • Hejun Wu
    • 1
  • Meiliang Li
    • 1
  • Xingyan Liu
    • 1
  • Anjun Chen
    • 1
  • Meng Ye
    • 2
  • Zhiqing Zhang
    • 1
    Email author
  1. 1.College of Food ScienceSichuan Agricultural UniversityYa’anChina
  2. 2.College of ForestrySichuan Agricultural UniversityChengduChina

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