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In Vivo Monitoring of Transcriptional Activity During Metabolic Transition Using a Bioluminescent Reporter in Yeast

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Luciferase detection during stationary phase in Lactococcus lactis.

Herwig Bachmann¹, Filipe Santos, Michiel Kleerebezem

¹NIZO food research, Kluyver Centre for Genomics of Industrial Fermentation, 6710 BA Ede, The Netherlands.

Applied and Environmental Microbiology

|May 22, 2007

Summary

Adding riboflavin or flavin mononucleotide to stationary-phase Lactococcus lactis improves detection of bacterial luciferase. This enhances the use of luciferase reporters in less metabolically active bacteria.

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Area of Science:

Microbiology
Biochemistry

Background:

Bacterial luciferase (luxAB) reporter systems are limited to metabolically active cells due to signal dependence on host metabolism.
Stationary-phase bacteria exhibit reduced metabolic activity, hindering reliable luciferase-based detection.

Purpose of the Study:

To improve the detection of bacterial luciferase in metabolically restricted, stationary-phase Lactococcus lactis.
To overcome limitations of current reporter systems in non-ideal bacterial growth phases.

Main Methods:

Whole-cell assay systems were utilized for Lactococcus lactis.
Addition of riboflavin and flavin mononucleotide was tested to enhance luminescence signals.

Main Results:

Detection of luciferase was significantly improved in stationary-phase Lactococcus lactis.
Supplementation with riboflavin or flavin mononucleotide enhanced the luminescence signal.

Conclusions:

Riboflavin and flavin mononucleotide can be added to improve bacterial luciferase detection in stationary-phase cells.
This finding expands the utility of luxAB reporter systems to a wider range of bacterial metabolic states.