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Abstract
A broad spectrum of volatile organic compounds’ (VOCs’) biological activities has attracted significant scientific interest, but their mechanisms of action remain little understood. The mechanism of action of two VOCs—the cyclic monoterpenes (−)-limonene and (+)-α-pinene—on bacteria was studied in this work. We used genetically engineered Escherichia coli bioluminescent strains harboring stress-responsive promoters (responsive to oxidative stress, DNA damage, SOS response, protein damage, heatshock, membrane damage) fused to the luxCDABE genes of Photorhabdus luminescens. We showed that (−)-limonene induces the PkatG and PsoxS promoters due to the formation of reactive oxygen species and, as a result, causes damage to DNA (SOSresponse), proteins (heat shock), and membrane (increases its permeability). The experimental data indicate that the action of (−)-limonene at high concentrations and prolonged incubation time makes degrading processes in cells irreversible. The effect of (+)-α-pinene is much weaker: it induces only heat shock in the bacteria. Moreover, we showed for the first time that (−)-limonene completely inhibits the DnaKJE–ClpB bichaperone-dependent refolding of heat-inactivated bacterial luciferase in both E. coli wild type and mutant ΔibpB strains. (+)-α-Pinene partially inhibits refolding only in ΔibpB mutant strain.
Highlights
In recent years essential oils have attracted significant scientific interest because they exhibit a broad spectrum of bioactivities, such as antibacterial, antifungal, antiviral, and insecticidal activities [1,2,3]
An increase in permeability and degradation of the cell membrane of Gram-negative E. coli and Gram-positive S. aureus bacteria were shown when they were incubated in a nutrient medium containing finger citron essential oil (FCEO) with limonene (~50%) and α-pinene as the main components [13]
It was shown that cyclic terpene limonene, when present in the growth medium during incubation for several hours, destroys the cell membrane in yeast cells (Z. rouxii) and bacterial cells (E. coli and S. aureus), which is accompanied by leakage of nucleic acids and proteins [8,13]
Summary
In recent years essential oils have attracted significant scientific interest because they exhibit a broad spectrum of bioactivities, such as antibacterial, antifungal, antiviral, and insecticidal activities [1,2,3] Their main active compounds—aldehydes, terpenes, and phenols—are widely used according to the recommendations of the US Food and. Volatile organic compounds (VOCs) limonene and pinene are of great interest in terms of practical application They are widespread in plants: lemon and other citrus fruits contain limonene, and the main active compound of essential oil obtained from conifers is pinene. An increase in permeability and degradation of the cell membrane of Gram-negative E. coli and Gram-positive S. aureus bacteria were shown when they were incubated in a nutrient medium containing finger citron essential oil (FCEO) with limonene (~50%) and α-pinene as the main components [13]. These results are obtained using genetically engineered Escherichia coli strains harboring stress-responsive promoters fused to the bioluminescent reporter gene system of lux operon
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