State Research Institute of Genetics and Selection of Industrial Microorganisms of the National Research Centre "Kurchatov Institute", Kurchatov Genomic Center, 117545 Moscow, Russia.
Institute of Molecular Genetics of the National Research Center "Kurchatov Institute", 123182 Moscow, Russia.
Biomolecules. 2021 May 29;11(6):806. doi: 10.3390/biom11060806.
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 bioluminescent strains harboring stress-responsive promoters (responsive to oxidative stress, DNA damage, SOS response, protein damage, heatshock, membrane damage) fused to the genes of We showed that (-)-limonene induces the P and P 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 wild type and mutant Δ strains. (+)-α-Pinene partially inhibits refolding only in Δ mutant strain.
（一）具有广谱生物活性的挥发性有机化合物（VOCs）引起了科学界的极大关注，但它们的作用机制仍知之甚少。本工作研究了两种挥发性有机化合物（环状单萜烯（-）-柠檬烯和（+）-α-蒎烯）对细菌作用机制。我们使用了含有应激反应启动子（对氧化应激、DNA 损伤、SOS 反应、蛋白质损伤、热休克、膜损伤有反应）的基因工程生物发光菌株，这些启动子与 基因融合。我们发现（-）-柠檬烯由于形成活性氧而诱导 P 和 P 启动子，从而导致 DNA（SOS 反应）、蛋白质（热休克）和膜（增加其通透性）损伤。实验数据表明，（-）-柠檬烯在高浓度和长时间孵育下的作用使细胞内的降解过程不可逆。（+）-α-蒎烯的作用要弱得多：它只诱导细菌发生热休克。此外，我们首次表明（-）-柠檬烯完全抑制了野生型和突变型 Δ 菌株中热失活的细菌荧光素酶在 DnaKJE-ClpB 双伴侣蛋白依赖性重折叠过程，（+）-α-蒎烯仅在 Δ 突变株中部分抑制重折叠。
