The biological activity and mechanisms of action of the unsaturated ketone β-ionone, a volatile compound of significantinterest for biotechnology, medicine, and agriculture, were studied. Using specific lux biosensors basedon Escherichia coli MG1655, we found that β-ionone causes oxidative stress in E. coli cells by inducing expression from the PkatG and Pdps promoters, but not from the PsoxS promoter. The effects of β-ionone on the heat shock induction (expression from the PibpA and PgrpE promoters) and on DNA damage (expression from the PcolD and PdinI promoters, SOS response) in E. coli cells were significantly weaker. β-Ionone did not cause oxidative stress in the cells of the gram-positive bacterium Bacillus subtilis.

The paper reports the results of a large-scale testing of antibacterial textiles with extremely stable and long-lasting copper oxide coating. Using disk diffusion method, ICP-OES and specific lux biosensors it was shown that the coating does not leach copper ions into the environment. Laboratory experiments performed according to the ISO 20743 protocol showed high antibacterial activity of the produced coating, up to complete growth suppression for some strains. The long-term field tests were carried out in a tropical climate, at the Climate test station "Hoa Lac" (Hanoi city, Vietnam). The number of microorganisms on the textile materials remained within the range of 1-3% in comparison with the control sample for the entire duration of the field exposure (12 months).

The diversity of the biological activity of volatile organic compounds (VOCs), including unsaturated ketone β-ionone, promising pharmacological, biotechnological, and agricultural agent, has aroused considerable interest. However, the functional role and mechanisms of action of VOCs remain insufficiently studied. In this work, the response of bacterial cells to the action of β-ionone was studied using specific bioluminescent lux-biosensors containing stress-sensitive promoters. We determined that in Escherichia coli cells, β-ionone induces oxidative stress (PkatG and Pdps promoters) through a specific response mediated by the OxyR/OxyS regulon, but not SoxR/SoxS (PsoxS promoter). It has been shown that β-ionone at high concentrations (50 μM and above) causes a weak induction of the expression from the PibpA promoter and slightly induces the PcolD promoter in the E. coli biosensors; the observed effect is enhanced in the ΔoxyR mutants. This indicates the presence of some damage to proteins and DNA. β-Ionone was found to inhibit the bichaperone-dependent DnaKJE-ClpB refolding of heat-inactivated bacterial luciferase in E. coli wild-type and ΔibpB mutant strains. In the cells of the Gram-positive bacterium Bacillus subtilis 168 pNK-MrgA β-ionone does not cause oxidative stress. Thus, in this work, the specificity of bacterial cell stress responses to the action of β-ionone was shown.

Volatile compounds emitted by bacteria can play a significant role in interacting with microorganisms, plants, and other organisms. In this work, we studied the effect of total gaseous mixtures of organic as well as inorganic volatile compounds (VCs) and individual pure volatile organic compounds (VOCs: ketones 2-nonanone, 2-heptanone, 2-undecanone, a sulfur-containing compound dimethyl disulfide) synthesized by the rhizosphere Pseudomonas chlororaphis 449 and Serratia plymuthica IC1270 strains, the soil-borne strain P. fluorescens B-4117, and the spoiled meat isolate S. proteamaculans 94 strain on Arabidopsis thaliana plants (on growth and germination of seeds). We demonstrated that total mixtures of volatile compounds emitted by these strains grown on Luria–Bertani agar, Tryptone Soya Agar, and Potato Dextrose Agar media inhibited the A. thaliana growth. When studied bacteria grew on Murashige and Skoog (MS) agar medium, volatile mixtures produced by bacteria could stimulate the growth of plants. Volatile compounds of bacteria slowed down the germination of plant seeds; in the presence of volatile mixtures of P. fluorescens B-4117, the seeds did not germinate. Of the individual VOCs, 2-heptanone had the most potent inhibitory effect on seed germination. We also showed that the tested VOCs did not cause oxidative stress in Escherichia coli cells using specific lux-biosensors. VOCs reduced the expression of the lux operon from the promoters of the katG, oxyS, and soxS genes (whose products involved in the protection of cells from oxidative stress) caused by the action of hydrogen peroxide and paraquat, respectively.

Bacteria and fungi emit a huge variety of volatile organic compounds (VOCs) that can provide a valuable arsenal for practical use. However, the biological activities and functions of the VOCs are poorly understood. This work aimed to study the action of individual VOCs on the bacteria Agrobacterium tumefaciens, Arabidopsis thaliana plants, and fruit flies Drosophila melanogaster. VOCs used in the work included ketones, alcohols, and terpenes. The potent inhibitory effect on the growth of A. tumefaciens was shown for 2-octanone and isoamyl alcohol. Terpenes (−)-limonene and (+)-α-pinene practically did not act on bacteria, even at high doses (up to 400 µmol). 2-Butanone and 2-pentanone increased the biomass of A. thaliana at doses of 200–400 μmol by 1.5–2 times; 2-octanone had the same effect at 10 μmol and decreased plant biomass at higher doses. Isoamyl alcohol and 2-phenylethanol suppressed plant biomass several times at doses of 50–100 μmol. Plant seed germination was most strongly suppressed by isoamyl alcohol and 2-phenylethanol. The substantial killing effect (at low doses) on D. melanogaster was exerted by the terpenes and the ketones 2-octanone and 2-pentanone. The obtained data showed new information about the biological activities of VOCs in relation to organisms belonging to different kingdoms.

This paper reports the results of the large-scale field testing of composite materials with antibacterial properties in a tropical climate. The composite materials, based on a cotton fabric with a coating of metal oxide nanoparticles (TiO2 and/or ZnO), were produced using high-power ultrasonic treatment. The antibacterial properties of the materials were studied in laboratory tests on solid and liquid nutrient media using bacteria of different taxonomic groups (Escherichia coli, Chromobacterium violaceum, Pseudomonas chlororaphis). On solid media, the coatings were able to achieve a >50% decrease in the number of bacteria. The field tests were carried out in a tropical climate, at the Climate test station “Hoa Lac” (Hanoi city, Vietnam). The composite materials demonstrated long-term antibacterial activity in the tropical climate: the number of microorganisms remained within the range of 1–3% in comparison with the control sample for the duration of the experiment (3 months). Ten of the microorganisms that most frequently occurred on the surface of the coated textiles were identified. The bacteria were harmless, while the fungi were pathogenic and contributed to fabric deterioration. Tensile strength deterioration was also studied, with the fabrics coated with metal oxides demonstrating a better preservation of their mechanical characteristics over time, (there was a 42% tensile strength decrease for the reference non-coated sample and a 21% decrease for the sample with a ZnO + CTAB coating).

Volatile organic compounds (VOCs) emitted by bacteria play an important role in the interaction between microorganisms and other organisms. They can inhibit the growth of phytopathogenic microorganisms, modulate plant growth, and serve as infochemicals. Here, we investigated the effects of ketones, alcohols, and terpenes on the colony biofilms of plant pathogenic Agrobacterium tumefaciens strains and swimming motility, which can play an important role in the formation of biofilms. It was shown that 2-octanone had the greatest inhibitory effect on biofilm formation, acting in a small amount (38.7 g/m3). Ketone 2-butanone and unsaturated ketone β-ionone reduced the formation of biofilms at higher doses (145.2–580.6 and 387.1–1548.3 g/m3, respectively, up to 2.5–5 times). Isoamyl alcohol and 2-phenylethanol decreased the formation of biofilms at doses of 88.7 and 122.9 g/m3 by 1.7 and 5 times, respectively, with an increased effect at 177.4 and 245.9 g/m3, respectively. The agrobacteria cells in mature biofilms were more resistant to the action of ketones and alcohols. These VOCs also suppressed the swimming motility of agrobacteria; the radius of swimming zones decreased ~from 2 to 5 times. Terpenes (−)-limonene and (+)-α-pinene had no significant influence on the colony biofilms and swimming motility at the doses used. The results obtained represent new information about the effect of VOCs on biofilms and the motility of bacteria.

Many volatile compounds secreted by bacteria play an important role in the interactions of microorganisms, can inhibit the growth of phytopathogenic bacteria and fungi, can suppress or stimulate plant growth and serve as infochemicals presenting a new type of interspecies communication. In this work, we investigated the effect of total pools of volatile substances and individual volatile organic compounds (VOCs) synthesized by the rhizosphere bacteria Pseudomonas chlororaphis 449 and Serratia plymuthica IC1270, the soil-borne strain P. fluorescens B-4117 and the spoiled meat isolate S. proteamaculans 94 on Arabidopsis thaliana plants. We showed that total gas mixtures secreted by these strains during their growth on Luria-Bertani agar inhibited A. thaliana growth. Hydrogen cyanide synthesis was unnecessary for the growth suppression. A decrease in the inhibition level was observed for the strain P. chlororaphis 449 with a mutation in the gacS gene, while inactivation of the rpoS gene had no effect. Individual VOCs synthesized by these bacteria (1-indecene, ketones 2-nonanone, 2-heptanone, 2-undecanone, and dimethyl disulfide) inhibited the growth of plants or killed them. Older A. thaliana seedlings were more resistant to VOCs than younger seedlings. The results indicated that the ability of some volatiles emitted by the rhizosphere and soil bacteria to inhibit plant growth should be considered when assessing the potential of such bacteria for the biocontrol of plant diseases.

Heavy metals and arsenic are environmental pollutants and have a negative effect on human health. Volatile organic compounds (VOCs) synthesized by bacteria are able to modulate the growth and metabolism of pro- and eukaryotes, to function as signals for distance communication between organisms, and to influence the action of various stress agents. The effects of sulfur-containing dimethyl disulfide (DMDS) and ketones (2-undecanone, 2-nonanone, 2-heptanone, 2-pentanone, and 2-propanone) on expression from the promoters of the copA, zntA, and arsR genes (PcopA, PzntA, and ParsR) responding to stresses caused by heavy metal ions and arsenic have been studied using lux-biosensors containing inducible promoters of the listed genes and the lux-operon as a reporter. The degree of expression from the respective promoters was determined by the intensity of bioluminescence. The effects of 2-undecanone, 2-nonanone, 2-heptanone, and DMDS led to a lower expression of lux-operon, which is controlled by PcopA and ParsR. The pretreatment of biosensors with these VOCs decreased the induction of expression from promoters PzntA and ParsR under conditions of exposure to ZnSO4 and NaAsO2, respectively, in the presence of CuSO4; this effect was marked only for 2-undecanone. 2-Pentanone increased the expression of lux-reporter from PcopA and PzntA. Pretreatment with 2-pentanone led to a 80% increase in expression of the lux reporter from PzntA compared to the effect of ZnSO4 without 2-pentanone. The effects of VOCs on the expression from promoters of the genes responding to the stresses induced by copper, zinc, and arsenic are multidirectional and depend on VOC structure.

The bacteria Serratia proteamaculans 94 have a LuxI/LuxR type QS system consisting of AHL synthase SprI and the regulatory receptor SprR. We have previously shown that inactivation of the AHL synthase sprI gene resulted in an increase in the invasive activity of S. proteamaculans correlated with an increased bacterial adhesion. In the present work, the effects of inactivation of the S. proteamaculans receptor SprR are studied. Our results show that inactivation of the receptor sprR gene leads to an increase in bacterial invasion without any increase in their adhesion. On the other hand, inactivation of the sprR gene increases the activity of the extracellular protease serralysin. Inactivation of the QS system does not affect the activity of the pore-forming toxin ShlA and prevents the ShlA activation under conditions of a limited concentration of iron ions typical of the human body. While the wild type strain shows increased invasion in the iron-depleted medium, deletion of its QS system leads to a decrease in host cell invasion, which is nevertheless similar to the level of the wild type S. proteamaculans grown in the iron-rich medium. Thus, inactivation of either of the two component of the S. proteamaculans LuxI/LuxR-type QS system leads to an increase in the invasive activity of these bacteria through different mechanisms and prevents invasion under the iron-limited conditions.

Quorum Sensing (QS) system regulates gene expression in response to a change in the density of the bacterial population. Facultative pathogen Serratia proteamaculans 94 has a LuxI/LuxR type QS system consisting of regulatory protein SprR and AHL synthase SprI. Invasive activity of these bacteria appears at the stationary growth phase corresponding to a maximal density of the bacterial population in vitro. To evaluate the contribution of QS system of S. proteamaculans 94 to the regulation of invasive activity, in this work, S. proteamaculans SprI(-) mutant carrying the inactivated AHL synthase gene was used. Inactivation of the AHL synthase sprI gene resulted in a more than fourfold increase in the invasive activity of S. proteamaculans preceded by the increased adhesion of bacteria to the cell surface. This effect correlated with the increased expression of the outer membrane protein ompX gene and the decrease in the activity of intrabacterial protease protealysin, whose substrate is OmpX. The inverse correlation between activity of protealysin and bacterial invasion was also observed in the model experiments under the iron-limiting culture conditions. These results show that QS system regulates the S. proteamaculans invasion. This regulation can involve changes both in the protealysin activity and in the level of the ompX gene transcription. (c) 2021 Published by Elsevier Masson SAS on behalf of Institut Pasteur.

В последние годы способность микроорганизмов синтезировать летучие органические соединения (ЛОС) и изучение их функциональной роли привлекает активное внимание исследователей различного профиля, работающих как в фундаментальных, так и в прикладных областях биологии. Известно, что бактерии и грибы выделяют огромное количество ЛОС различной химической природы. Эти вещества характеризуются низкой молекулярной массой (в среднем порядка 300 Да), липофильной природой, низкой температурой кипения. Среди них – спирты, кетоны, терпены, пиразины, альдегиды, серо- и азот-содержащие соединения и др. In recent years, the ability of microorganisms to synthesize volatile organic compounds (VOCs) and the study of their functional role has attracted the active attention of researchers in various fields, working in both fundamental and applied fields of biology. It is known that bacteria and fungi emit a huge amount of VOCs of various chemical nature. These substances are characterized by low molecular weight (about 300 Da on average), lipophilic nature, and low boiling point. Among them are alcohols, ketones, terpenes, pyrazines, aldehydes, sulfur- and nitrogen-containing compounds, etc.

The quorum sensing (QS) system SprIR of the psychrotrophic strain Serratia proteamaculans 94 was investigated. A mutant was constructed with the inactivated s prR gene encoding the regulatory receptor protein SprR. Inactivation of this gene was shown to affect the composition of fatty acids synthesized by S. proteamaculans 94 and did not affect the synthesis of N -acyl-L-homoserine-lactones (AHL); the activities of extracellular proteases, chitinases, and hemolysins; the swimming motility of cells; and the suppression of mycelium growth of fungal plant pathogens by volatile compounds emitted by this strain. Inactivation of the sprI gene (but not the s prR gene) reduced the biofilm formation, which increased when exogenous AHL was added to the culture. The comparative proteomic analysis of cell of the parent strain and mutant strains with inactivated sprI and sprR genes showed that the expression of 30 proteins in S. proteamaculans 94 is affected by the SprIR quorum sensing system.

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.

Microbial volatile organic compounds (VOCs) are cell metabolites that affect many physiological functions of prokaryotic and eukaryotic organisms. Earlier we have demonstrated the inhibitory effects of soil bacteria volatiles, including ketones, on cyanobacteria. Cyanobacteria are very sensitive to ketone action. To investigate the possible molecular mechanisms of the ketone 2-nonanone influence on cyanobacterium Synechococcus elongatus PCC 7942, we applied a genetic approach. After Tn5-692 transposon mutagenesis, several 2-nonanone resistant mutants have been selected. Four different mutant strains were used for identification of the impaired genes (Synpcc7942_1362, Synpcc7942_0351, Synpcc7942_0732, Synpcc7942_0726) that encode correspondingly: 1) a murein-peptide ligase Mpl that is involved in the biogenesis of cyanobacteria cell wall; 2) a putative ABC transport system substrate-binding proteins MlaD, which participates in ABC transport system that maintains lipid asymmetry in the gram-negative outer membrane by aberrantly localized phospholipids transport from outer to inner membranes of bacterial cells; 3) a conserved hypothetical protein that is encoding by gene belonging to phage gene cluster in Synechococcus elongatus PCC 7942 genome; 4) a protein containing the VRR-NUC (virus-type replication-repair nuclease) domain present in restriction-modification enzymes involved in replication and DNA repair. The obtained results demonstrated that 2-nonanone may have different targets in Synechococcus elongatus PCC 7942 cells. Among them are proteins involved in the biogenesis and functioning of the cyanobacteria cell wall (Synpcc7942_1362, Synpcc7942_0351, Synpcc7942_0732) and protein participating in stress response at DNA restriction-modification level (Synpcc7942_0726). This paper is the first report about the genes that encode protein products, which can be affected by 2-nonanone.

The study provides new evidence for Ag-coated polyester (PES) mediating Escherichia coli inactivation by way of genetically engineered E. coli (without porins, from now denoted porinless bacteria). This allows the quantification of the bactericidal kinetics induced by the Ag surface without the intervention of Ag ions. Bacterial inactivation mediated by Ag-PES was seen to be completed within 60 min. The samples were prepared by high-power impulse magnetron sputtering (HiPIMS) at different sputter powers. In anaerobic media, this process required 120 min. The amounts of ions (Ar+, Ag+, and Ag2+) generated during the deposition by direct current magnetron sputtering (DCMS) and HiPIMS were determined by mass spectrometry. The thickness of the Ag films sputtered on PES by DCMS (0.28 A) during 100 s was found to be 340 nm. Thicknesses of 250, 230, and 200 nm were found when sputtering with HiPIMS was tuned at 8, 17, and 30 A, respectively. By scanning transmission electron microscopy (STEM-HAADF), the atomic distribution of Ag and oxygen was detected. By X-ray photoelectron spectroscopy (XPS), a shift in the Ag oxidation state was observed within the bacterial inactivation period. This reveals redox catalysis within the time required for the total bacterial inactivation due to the interaction between the bacterial suspension and Ag-PES. Surface properties of the Ag-coated PES samples were additionally investigated by X-ray diffraction (XRD). The formation of Ag plasmon was detected by diffuse reflectance spectroscopy (DRS) and was a function of the applied sputtering energy. The indoor sunlight irradiation dose required to induce an accelerated bacterial inactivation was found to be 5-10 mW/cm2.

In the present work, we provide evidence for visible light irradiation of the Au/TiO2 nanoparticles' surface plasmon resonance band (SPR) leading to electron injection from the Au nanoparticles to the conduction band of TiO2. The Au/TiO2 SPR band is shown to greatly enhance the light absorption of TiO2 in the visible region. Evidence is presented for the light absorption by the Au/TiO2 plasmon bands leading to the dissolution of Au nanoparticles. This dissolution occurs concomitantly with the injection of the hot electrons generated by the Au plasmon into the conduction band of TiO2. The electron injection from the Au nanoparticles into TiO2 was followed by femtosecond spectroscopy. The formation of Au ions was further confirmed by the spectral shift of the transient absorption spectra of Au/TiO2. The spectral changes of the SPR band of Au/TiO2 nanoparticles induced by visible light were detected by spectrophotometer, and the morphological transformation of Au/TiO(2 )was revealed by electron microscopy techniques as well. Subsequently, the fate of the Au ions was sorted out during the growth and biofilm formation for some selected Gram-negative bacteria. This study compares the bactericidal mechanism of Au ions and Ag ions, which were found to be substantially different depending on the selected cell used as a probe.

Microbial volatiles have a significant impact on the physiological functions of prokaryotic and eukaryotic organisms. Various ketones are present in volatile mixtures produced by plants, bacteria, and fungi. Our earlier results demonstrated the inhibitory effects of soil bacteria volatiles, including ketones, on cyanobacteria. In this work, we thoroughly examined the natural ketones, 2‐nonanone and 2‐undecanone to determine their influence on the photosynthetic activity in Synechococcus sp. PCC 7942. We observed for the first time that the ketones strongly inhibit electron transport through PSII in cyanobacteria cells in vivo. The addition of ketones decreases the quantum yield of primary PSII photoreactions and changes the PSII chlorophyll fluorescence induction curves. There are clear indications that the ketones inhibit electron transfer from QA to QB, electron transport at the donor side of PSII. The ketones can also modify the process of energy transfer from the antenna complex to the PSII reaction center and, by this means, increase both chlorophyll fluorescence quantum yield and the chlorophyll excited state lifetime. At the highest tested concentration (5 mM) 2‐nonanone also induced chlorophyll release from Synechococcus cells that strongly indicates the possible role of the ketones as detergents.