Vol. 10 No. 4 (2023)

Heterologous Expression and Partial Purification of Plantaricin Produced by Lactiplantibacillus plantarum COY2906

Applied Food Biotechnology, Vol. 10 No. 4 (2023), 25 September 2023
https://doi.org/10.22037/afb.v10i4.42761

Abstract

Background and Objective: Lactiplantibacillus plantarum COY2906 was isolated from virgin coconut oil, a strain known for its production of plantaricin which acts as a bio-preservative. The aim of this study was to investigate specific plantaricin genes of plnA, plnEF, plnN, plnJ and plnK, precipitate the plantaricin with ammonium sulfate and assess antimicrobial activity of the crude plantaricin.

Material and Methods: Growth analysis of strain COY2906 was monitored using spectrophotometer. Amplification and detection of gene targets were carried out using real-time polymerase chain reaction (Real-Time PCR). Crude plantaricin was assessed using 40 and 70% (w/v) ammonium sulphate. Antimicrobial activity was assessed using well-diffusion assay and the molecular mass of partially purified protein was assessed using matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF-MS).

Results and Conclusion: The plantarum strain COY2906 was cultured in MRS broth at 37 °C under anaerobic conditions and harvested after 19 h or in the middle of the stationary phase to maximize production of plantaricin. Relative expression level of plnA, plnEF, plnN and plnJ were over-expressed, while that of plnK was not. To achieve plantaricin, cell-free supernatant was precipitated with 40 and 70% ammonium sulphate, resulting in crude protein concentrations of 41.33 and 148 µg.ml-1, respectively. Crude protein had no antimicrobial activities, cell-free supernatant of the strain COY2906 showed a comparable antimicrobial efficacy to that of sodium ampicillin at 100 µg.ml-1. Matrix-assisted laser desorption ioniza-tion mass spectrometry spectrum did not show the presence of plantaricin A, plantaricin EF, plantaricin N and plantaricin J after precipitation with 70% ammonium sulphate. However, plantaricin K was detected in the spectrum. Regarding the results, further analysis on the detection of plantaricin is recommended using matrix-assisted laser desorption ionization mass spectrometry. This may involve modifying the solvent or increasing concentration of ammonium sulphate to assess its activities and characteristics.

Conflict of interest: The authors declare no conflict of interest.

Keywords:
  • Antimicrobial Activity
  • Crude Protein
  • Gene Expressions
  • L. plantarum
  • Plantaricin

How to Cite

Syaputri, Y., Lei, J., Ratningsing, N., Rossiana, N., Safitri, R., & Wulandari, A. P. (2024). Heterologous Expression and Partial Purification of Plantaricin Produced by Lactiplantibacillus plantarum COY2906. Applied Food Biotechnology, 10(4). https://doi.org/10.22037/afb.v10i4.42761 (Original work published October 9, 2023)

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