Bright stable luminescent yeast using bacterial luciferase as a sensor

Area of Science:

• Biotechnology
• Molecular Biology
• Biochemistry

Background:

• Bacterial luciferase (LuxAB) enables bioluminescence but requires specific substrates.
• Optimizing substrate delivery and enzyme activity is crucial for efficient light emission in eukaryotic systems.
• Previous studies lacked efficient and non-toxic substrate systems for yeast bioluminescence.

Purpose of the Study:

• To engineer yeast cells capable of bright and sustained bioluminescence using bacterial luciferase.
• To evaluate the efficacy and toxicity of different fatty aldehyde substrates for yeast bioluminescence.
• To enhance bioluminescence by increasing the availability of reduced flavin mononucleotide (FMNH2).

Main Methods:

• Introduction of bacterial luciferase (LuxAB) genes into yeast.
• Supplementation with saturating, non-toxic levels of bioluminescence substrates, including Z-9-tetradecenal and decanal.
• Transformation of yeast with the NADPH-specific FMN reductase (FRP) gene from Vibrio harveyi.

Main Results:

• Yeast cells expressing LuxAB achieved light intensities comparable to bacterial systems.
• Z-9-tetradecenal supported strong, sustained luminescence (>24h) and was non-toxic at tested concentrations, unlike decanal.
• Co-expression of FMN reductase (FRP) resulted in a 100-fold increase in luminescence intensity.

Conclusions:

• Z-9-tetradecenal is a highly effective and non-toxic substrate for bacterial luciferase-mediated bioluminescence in yeast.
• Enhancing FMNH2 regeneration via FMN reductase significantly boosts light output in engineered yeast.
• Bacterial luciferase holds strong potential as a light-emitting sensor in eukaryotic organisms, particularly yeast.