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Quantification of Bacillus thuringiensis Vip3Aa16 Entomopathogenic Toxin Using Its Hemolytic Activity
Hanen Boukedi1, Saoussen Ben Khedher1, Dhouha Ghribi2
1Laboratory of Biopesticides, Centre of Biotechnology of Sfax, University of Sfax, P.O. Box 1177, 3018, Sfax, Tunisia.
Current Microbiology
|March 6, 2017
Summary
Quantifying Bacillus thuringiensis Vip3Aa16 toxin is now simpler. This study shows Vip3Aa16 induces hemolysis, a stable and measurable activity, enabling easy biopesticide assessment for pest management.
Area of Science:
- Agricultural Science
- Biochemistry
- Microbiology
Background:
- Bacillus thuringiensis (Bt) produces insecticidal proteins vital for biopesticides.
- Quantifying secreted Bt toxins, like Vip3Aa16, has been a significant challenge.
- Effective pest management relies on accurate biopesticide quantification.
Purpose of the Study:
- To develop a simple and credible method for quantifying Vip3A toxins.
- To investigate the hemolytic activity of the Vip3Aa16 toxin.
- To establish a link between hemolysis and Vip3A toxin concentration.
Main Methods:
- Heterologous expression of Vip3Aa16 toxin in Escherichia coli.
- Purification of Vip3Aa16 protein using chromatography.
- Assessment of hemolytic activity using blood agar and zymogram assays.
Main Results:
- Vip3Aa16 was shown to induce hemolysis, a novel finding.
- Hemolytic activity remained stable at 40°C and pH 6.5-9.
- A linear correlation was established between hemolysis levels and Vip3Aa16 concentration.
Conclusions:
- A simple, low-cost hemolysis-based assay can quantify Vip3A toxins.
- This method facilitates the assessment of Vip3A toxins in liquid cultures.
- The assay supports the integration of Vip3A biopesticides into pest management programs.
