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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
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Making plants into cost-effective bioreactors for highly active antimicrobial peptides
Meron Ghidey1, S M Ashiqul Islam1, Grace Pruett2
1Biomedical Studies Program, Baylor University, Waco, TX, 76798, USA.
New Biotechnology
|December 9, 2019
Summary
Plant expression of antimicrobial peptides (AMPs) can be enhanced by reducing their positive charge. This study achieved high yields of active AMP-elastin-like polypeptide fusions in plants, offering a new strategy against antibiotic resistance.
Area of Science:
- Biotechnology
- Molecular Biology
- Plant Science
Background:
- Antibiotic-resistant pathogens are a growing global threat.
- Antimicrobial peptides (AMPs) show promise as alternatives to conventional antibiotics.
- Current plant-based expression systems for AMPs yield low levels and can cause cytotoxicity.
Purpose of the Study:
- To investigate the impact of AMP charge on plant expression efficiency.
- To develop a high-yield plant expression system for functional AMPs.
- To optimize AMP purification and enhance antimicrobial activity.
Main Methods:
- Meta-analysis of protein databases to compare AMP cationicity across taxa.
- Transient expression of 10 heterologous AMPs fused to elastin-like polypeptide (ELP) in *Nicotiana benthamiana*.
- Temperature-shift purification of ELP-AMP fusions and assessment of antimicrobial activity.
Main Results:
- Native plant AMPs exhibit significantly lower cationicity than those from other organisms.
- Anionic AMPs expressed efficiently in plants, with two achieving exceptionally high yields (375 and 563 μg/gfw).
- ELP-AMP fusions demonstrated enhanced antimicrobial activity against *Staphylococcus epidermidis* (MIC of 0.26 μM) without cleavage.
Conclusions:
- Reducing AMP cationicity is crucial for efficient expression in plants.
- ELP fusion provides a cost-effective method for high-yield production and purification of active AMPs in plants.
- This plant-based expression system offers a viable strategy for producing potent antimicrobial agents to combat antibiotic resistance.
