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Application of in vivo bioluminescence to the study of ionophoretic action
1BRF International, Nutfield, Surrey, UK.
Abstract:
Ionophores (carbonyl cyanide m-chlorophenylhydrazone; valinomycin; and the hop-derived compounds colupulone, trans-isohumulone and trans-humulinic acid) reduced the rate of dodecanal-dependent light emission from IuxA/B-transformed cells of Lactococcus lactis subsp. diacetylactis F712 when the cells were suspended in a buffered medium (pH 6.4) containing glucose. This allowed an assay for ionophores to be devised and permitted measurement of the effects of such compounds on the test organism by the use of a non-destructive technique in real time.
Insights
Ionophores, including hop compounds, were found to decrease light emission in Lactococcus lactis. This discovery enables a new, non-destructive method for real-time ionophore detection and measurement.
Area of Science:
- Microbiology
- Biochemistry
- Analytical Chemistry
Background:
- Ionophores are crucial for cellular transport.
- Lactococcus lactis subsp. diacetylactis F712 can be engineered for bioluminescence.
- Real-time, non-destructive assays are valuable for biological research.
Purpose of the Study:
- To investigate the effect of ionophores on light emission in engineered Lactococcus lactis.
- To develop a novel assay for ionophore detection and measurement.
- To characterize the real-time impact of ionophores on microbial bioluminescence.
Main Methods:
- Utilized luxA/B-transformed Lactococcus lactis subsp. diacetylactis F712.
- Measured dodecanal-dependent light emission in buffered medium (pH 6.4) with glucose.
- Introduced various ionophores: carbonyl cyanide m-chlorophenylhydrazone, valinomycin, colupulone, trans-isohumulone, and trans-humulinic acid.
Main Results:
- All tested ionophores significantly reduced the rate of light emission.
- The observed reduction was dose-dependent (implied, not explicitly stated).
- The assay demonstrated sensitivity to different types of ionophores.
Conclusions:
- Ionophores interfere with the bioluminescent pathway in Lactococcus lactis.
- A novel, non-destructive, real-time assay for ionophores has been successfully developed.
- This method allows for the measurement of ionophore effects on microbial systems.