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Establishing Heterologous Production of Microcins J25 and Y in Bacillus subtilis
Guangwen Zhang1, Min Lin1, Miaomiao Qin1
1College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P. R. China.
Abstract:
Microcin J25 (MccJ25) and microcin Y (MccY) are lasso peptides and considered potential alternatives to antibiotics and harmful preservatives. The combination of these two microcins can provide a wide antimicrobial spectrum against food-borne Salmonella. Currently, MccJ25 and MccY are produced using Escherichia coli expression systems; however, the entire production process is accompanied by negative effects from endotoxins. In this study, we identified Bacillus subtilis as a suitable host for MccJ25 and MccY production. High-level production of microcins was achieved by promoter optimization, host strain selection, and recombinant expression. The engineered strains produced maximum yields of 2.827 μM MccJ25 and 1.481 μM MccY. This is the first study to demonstrate the expression of MccJ25 and MccY in B. subtilis, and it offers a few engineered strains that are without antibiotic resistance markers, inducer-free, sporulation-deficient, and free of the negative effects of endotoxins for antibacterial therapy and food preservation.
Insights
This study successfully produced microcin J25 (MccJ25) and microcin Y (MccY) in Bacillus subtilis, offering a safer alternative to traditional methods. These engineered strains provide endotoxin-free antimicrobial solutions for food preservation and antibacterial therapy.
Area of Science:
- Biotechnology
- Microbiology
- Molecular Biology
Background:
- Microcin J25 (MccJ25) and microcin Y (MccY) are lasso peptides with potential as antimicrobial agents.
- Current production methods using Escherichia coli are limited by endotoxin contamination.
- A safe and efficient production system is needed for MccJ25 and MccY.
Purpose of the Study:
- To establish Bacillus subtilis as a viable host for MccJ25 and MccY production.
- To develop engineered B. subtilis strains for high-level microcin yield.
- To create endotoxin-free microcin production for therapeutic and food preservation applications.
Main Methods:
- Engineered Bacillus subtilis strains for recombinant expression of MccJ25 and MccY.
- Optimized promoter sequences and selected host strains for enhanced production.
- Evaluated microcin yields and characterized engineered strains for desirable traits.
Main Results:
- Successfully expressed MccJ25 and MccY in B. subtilis.
- Achieved maximum yields of 2.827 μM MccJ25 and 1.481 μM MccY.
- Developed engineered strains that are inducer-free, sporulation-deficient, and lack antibiotic resistance markers.
Conclusions:
- Bacillus subtilis is a suitable host for producing MccJ25 and MccY.
- The developed strains offer an endotoxin-free production platform.
- This research provides engineered strains for safe antibacterial therapy and food preservation.
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