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Influence of PepF peptidase and sporulation on microcin J25 production in Bacillus subtilis

Guangwen Zhang1, Saixiang Feng1,2,3,4,5, Miaomiao Qin1

  • 1College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.

Microbiology Spectrum
|May 23, 2024
PubMed

Insights

Microcin J25 (MccJ25) production in Bacillus subtilis is enhanced by deleting the peptidase PepF and modifying sporulation pathways. This strategy significantly increases MccJ25 yield and stability for therapeutic and preservative applications.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Microcin J25 (MccJ25) is a lasso peptide with potent antibacterial properties, making it a promising candidate for drug development and food preservation.
  • Previous work established heterologous expression of MccJ25 in Bacillus subtilis, but its regulation and accumulation mechanisms remained unclear.
  • Understanding factors limiting MccJ25 production is crucial for optimizing its yield and therapeutic potential.

Purpose of the Study:

  • To investigate the role of peptidases PepA, PepF, and PepT in the regulation and stability of MccJ25 production in B. subtilis.
  • To identify factors limiting MccJ25 accumulation and develop strategies for enhanced production.
  • To engineer B. subtilis strains for improved MccJ25 yield and stability.

Main Methods:

  • Investigated MccJ25 expression levels in B. subtilis strains with disrupted peptidase genes (pepA, pepF, pepT).
  • Analyzed MccJ25 concentration in pepF mutants compared to wild-type strains.
  • Examined the effect of MccJ25 production on sporulation-associated genes in B. subtilis.

Main Results:

  • Oligoendopeptidase F (PepF) was identified as a key enzyme degrading the MccJ25 precursor peptide, thus reducing production.
  • The pepF mutant strain achieved a significantly higher MccJ25 concentration (1.68 µM) compared to the wild-type (0.14 µM) after extended cultivation.
  • MccJ25 production was found to downregulate sporulation-associated genes, potentially contributing to its accumulation.

Conclusions:

  • Deletion of the pepF gene and modulation of sporulation pathways are effective strategies to enhance MccJ25 production and stability in B. subtilis.
  • Engineered B. subtilis strains exhibit significantly improved MccJ25 yields, reducing production costs and expanding utilization possibilities.
  • This study provides a foundation for optimizing MccJ25 as a therapeutic agent and safe food preservative.
Keywords:
Bacillus subtilisPepFmicrocin J25peptidasesporulation

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