Dynamic D-Allose Monitoring Via a Toggle-Programmed Microbial Fluorescent Biosensor

Area of Science:

• Biotechnology
• Microbiology
• Synthetic Biology

Background:

• Rare sugars like d-allose are valuable in food and pharmaceuticals.
• Current d-allose detection relies on complex and time-consuming chromatography.
• Need for efficient monitoring methods in microbial fermentation.

Purpose of the Study:

• To develop a novel microbial fluorescent biosensor for dynamic d-allose monitoring.
• To enable real-time detection of d-allose in fermentation broths.
• To provide a simpler and more cost-effective alternative to chromatography.

Main Methods:

• Characterization of the d-allose-sensing P_alsR promoter.
• Construction of a toggle-programmed biosensor using P_T7 and P_alsR promoters for GFP and T7 RNAP expression.
• Overexpression of AlsR repressor using P_pdc promoter for P_alsR regulation.
• Utilized mGFP-DAS as a fast-response reporter molecule.

Main Results:

• The biosensor demonstrated high sensitivity to d-allose.
• Measurement deviation was controlled within 15%.
• The use of mGFP-DAS facilitated rapid detection during continuous incubation.

Conclusions:

• The developed microbial fluorescent biosensor is a viable tool for d-allose detection.
• This method offers significant advantages in simplicity and cost-effectiveness over chromatography.
• Enables dynamic monitoring of rare sugars in microbial fermentation.