Abstract

Most of the probiotics Bacterial cells, express native antibacterial genes, resulting in the production of, antimicrobial peptides, which have various applications in biotechnology and drug development. But the identification of antibacterial peptide, structural characterization of antimicrobial peptide and prediction on mode of action. Regardless of the significance of protein manufacturing, three individual factors are required for the production method: gene expression, stabilization and specific peptide purification. Our protocol describes a straightforward technique of detecting and characterizing particular extracellular peptides and enhancing the antimicrobial peptide expression we optimized using low molecular weight peptides. This protocol can be used to improve peptide detection and expression. The following are the benefits of this method, (DOI – https://doi.org/10.1016/j.ijbiomac.2019.10.196 [1]).The data briefly describe a simple method in detection identification, characterization of antimicrobial extracellular peptide, predicating the mode of action of peptide in targeting pathogens (In-silico method), brief method on profiling of antimicrobial peptide and its mode of action [1]. Further the protocol can be used to enhance the specific peptide expressions, detection of peptides. The advantages of this technique are presented below:•Characterization protocol of specific antimicrobial peptide•The folded antimicrobial peptide expression were less expressed or non-expressed peptides.•Besides being low cost, less time-consuming, easy to handle, universal and fast to execute, the suggested technique can be used for multiple proteins expressed in probiotics (Lactobacillus species) expression system.

Highlights

  • Most of the probiotics Bacterial cells, express native antibacterial genes, resulting in the production of, antimicrobial peptides, which have various applications in biotechnology and drug development

  • Bacterial progression condition The 2% of Pediococcus strain, were inoculated in De Man, Rogosa and Sharpe (MRS) broth and incubated at 37 ± 2°C for 24 h, further incubated samples were aseptically transferred into sterile round bottom centrifuge tubes and centrifuged at 4000g for 15 min and cell free supernatant (CFS) was collected and proceeded for purification of bioactive compound

  • The size exclusion spin column chromatography,

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Summary

Data accessibility Related research article

All data are presented in this article and raw data were shared through file share.com https://figshare.com/s/a6bd074 8a4 8581faf3ce. Bacterial progression condition The 2% of Pediococcus strain, were inoculated in De Man, Rogosa and Sharpe (MRS) broth and incubated at 37 ± 2°C for 24 h, further incubated samples were aseptically transferred into sterile round bottom centrifuge tubes and centrifuged at 4000g for 15 min and cell free supernatant (CFS) was collected and proceeded for purification of bioactive compound

Purification of bioactive compound
Identification of peptides by mass spectrometry
Bradford protein assay
Precipitation and identification of secreted proteins
Tricine SDS-PAGE Gel electrophoreses
Tricine-Gel fraction and elution
Swiss-model: homology modelling of protein structures
Findings
In silico – molecular interaction and docking

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