Fermentation Revisited: How Do Microorganisms Survive Under Energy-Limited Conditions?

During fermentation FOF1 hydrolyzes ATP, coupling proton transport to proton-motive force (pmf) generation. Despite that, pmf generated by ATP hydrolysis does not satisfy the energy budget of a fermenting cell. However, pmf can also be generated by extrusion of weak organic acids such as lactate and by hydrogen cycling catalyzed by hydrogenases (Hyds). Here we highlight recent advances in our understanding of how the transport of weak organic acids and enzymes contributes to pmf generation during fermentation. The potential impact of these processes on metabolism and energy conservation during microbial fermentation have been overlooked and they not only expand on Mitchell's chemiosmotic theory but also are of relevance to the fields of microbial biochemistry and human and animal health.

Anti-cancer effect of in vivo inhibition of nitric oxide synthase in a rat model of breast cancer.

Increased expression of nitric oxide synthase (NOS) is associated with different cancers such as cervical, breast, lung, brain, and spinal cord. Inhibition of NOS activity has been suggested as potential tool to prevent breast cancer. The anti-tumor therapeutic effect of L-nitro arginine methyl ester (L-NAME), NOS inhibitor, using in vivo models is currently under investigation. We hypothesized that L-NAME will show an anti-tumor effect by delaying a progression of breast cancer via a modulation of cell death and proliferation, and angiogenesis. We used a novel model of anti-cancer treatment by the administration of L-NAME (30 mg/kg in a day, intraperitoneal) injected every third day for five weeks to rat model of 7,12-dimethylbenz[a]anthracene (DMBA)-induced breast tumor. Concentrations of nitrite anions, polyamines, malondialdehyde, NH4+ levels, and arginase activity in the blood were decreased in DMBA + L-NAME-treated rats compared with DMBA rats. The mortality rates, tumor number, weight, and volume, as well as the histopathological grade of breast cancer were also significantly reduced. In addition, L-NAME treatment showed a delay in tumor formation, and in body weight compared with rats administrated only with DMBA. In conclusion, our data show that L-NAME is a promising anti-cancer agent to treat breast cancer, which can lead to development of anti-tumor therapeutic tools in future.

Changes in growth kinetic parameters, morphology and mitotic activity of yeasts Candida guilliermondii exposed to the low-intensity waves of 51.8-GHz frequency.

Under the influence of electromagnetic waves of millimeter range with the frequency of 51.8 GHz, changes in the morphology, growth parameters and mitotic activity of yeasts C. guilliermondii NP-4 are revealed. Filamentous and giant cells appeared in a population of exposed yeasts. The sigmoid shape of the growth curve remained but the lag phase duration was increased by 2 h in comparison with non-exposed yeasts; accordingly, the log and stationary phases followed 2 h later. The specific growth rate in the log growth phase and colony-forming ability of exposed yeasts was decreased. It is suggested that yeasts have some response mechanisms to 51.8-GHz frequency electromagnetic waves. The results can be used to understand the response mechanisms of microorganisms to non-ionizing radiation, as well as to develop approaches to protect living organisms from it. The effect of electromagnetic waves of 51.8-GHz frequency to suppress yeasts can be applied in biotechnology and medicine.

Formate and potassium ions affect Escherichia coli proton ATPase activity at low pH during mixed carbon fermentation.

Escherichia coli is able to ferment not only single but also mixtures of carbon sources. The formate metabolism and effect of formate on various enzymes have been extensively studied during sole glucose but not mixed carbon sources utilization. It was revealed that in membrane vesicles (MV) of wild type cells grown at pH 7.5 during fermentation of the mixture of glucose (2 g/L), glycerol (10 g/L), and formate (0.68 g/L), in the assays, the addition of formate (10 mM) increased the N,N'-dicyclohexylcarbodiimide (DCCD)-inhibited ATPase activity on ~30% but no effect of potassium ions (100 mM) had been detected. In selC (coding formate dehydrogenases) and fdhF (coding formate dehydrogenase H) single mutants, formate increased DCCD-inhibited ATPase activity on ~40 and ~70%, respectively. At pH 5.5, in wild type cells MV, formate decreased the DCCD-inhibited ATPase activity ~60% but unexpectedly in the presence of potassium ions, it was stimulated ~5.8 fold. The accessible SH or thiol groups number in fdhF mutant was less by 28% compared with wild type. In formate assays, the available SH groups number was less ~10% in wild type but not in fdhF mutant. Taken together, the data suggest that proton ATPase activity depends on externally added formate in the presence of potassium ions at low pH. This effect might be regulated by the changes in the number of redox-active thiol groups via formate dehydrogenase H, which might be directly related to proton ATPase FO subunit.

Influence of C4 -Dcu transporters on hydrogenase and formate dehydrogenase activities in stationary phase-grown fermenting Escherichia coli.

During mixed-acid fermentation, Escherichia coli transports succinate mainly via transporters of the Dcu family. Here, we analyze the influence of Dcu transporters on hydrogenase (Hyd) and fermentative formate dehydrogenase (FDH-H) activities and how this is affected by external pH and carbon source. Using selected dcu mutations, it was shown that Dcu carriers mainly affect Hyd and FDH-H activities during glycerol but not glucose fermentation at acidic pH. During glycerol fermentation at pH 5.5, inactivation of either one or all Dcu carriers increased total Hyd activity by 60% compared with wild type. Under the same growth conditions, a dcuACBD mutant had a twofold higher FDH-H activity. When glucose was fermented in dcuD single mutant at pH 5.5, the FDH-H activity was also increased twofold compared with wild type. Interestingly, in dcuD or dcuACBD mutants at pH 7.5, Hyd activity was lowered by 20%. Taken together, it can be concluded that during glucose fermentation at pH 7.5, lack of DcuD affects Hyd enzyme activity, but at pH 5.5, it has a stronger effect on FDH-H activity. During glycerol fermentation, lack of Dcu carriers increased Hyd and FDH-H activities as revealed at pH 5.5. The results suggest that impairing Dcu transport function increases intracellular formate levels and thus affects H2 cycling and proton-motive force generation.

Phytochemical Screening and Detection of Antibacterial Components from Crude Extracts of Some Armenian Herbs Using TLC-Bioautographic Technique.

The aim of the research was to explore qualitative and quantitative phytochemical constituents of different extracts from Agrimonia eupatoria L., Hypericum alpestre subsp. polygonifolium (Rupr.) Avet. & Takht., Rumex obtusifolius Willd and Sanguisorba officinalis L. and analyze their bioactive constituents using TLC-bioautography technique. In TLC analysis different solvent systems as mobile phases were used in order to make assumptions about the nature of active antibacterial compounds. The highest total phenolic, flavonoid and tannin content was detected in methanol extract of A. eupatoria (358.9 µg Gallic acid equivalent per mg dry weight), acetone extract of H. alpestre (216.75 µg Quercetin equivalent per mg dry weight) and acetone extract of R. obtusifolius (76.24 µg Catechin equivalent per mg dry weight), respectively. TLC-bioautography analysis using mobile phases with different polarity allowed to separate different bands from tested plants methanol and acetone extracts possessing antibacterial activity against Staphylococcus aureus MDC 5233. Thus, it was revealed, that polar compounds could have considerable contribution to the antibacterial activity of acetone and methanol extracts of H. alpestre and R. obtusifolius. It was hypothesized, that complex compounds can be responsible for the antibacterial action of extracts of A. eupatoria and S. officinalis. Based on obtained data, A. eupatoria, H. alpestre, R. obtusifolius and S. officinalis plants can be recommended as sources of new antibacterial agents.

Antifungal Activity of Lactic Acid Bacteria Isolates and Their Associations: The Effects of Ca and Mg Divalent Cations.

The study of effects of Ca2+ and Mg2+ on antifungal activity of lactic acid bacteria (LAB) isolates and their associations revealed inducing and inhibiting effects on antifungal activity. The addition of Ca2+ essentially inhibited the antifungal effect of L. rhamnosus MDC9661 but stimulated the activity of RIN-2003-Ls, MDC9632 and MDC9633 strains, as well as their associations. Mg2+ partly increased the inhibitory activity of LAB isolates, while the addition of ions combination did not cause changes of their antifungal activity. The supplementation of Ca2+ stimulated the antifungal effect of most associations against Penicillium sp., Trichoderma viride, Geotrichum candidum, and Aspergillus flavus compared with the native conditions. The addition of Mg2+ induced the antifungal activity of RIN-2003-Ls, MDC9632, MDC9633, and INR-2010-Tsov-G-St combinations. The antifungal effects of most associations were increased in the presence of ions mixture. The natural LAB associations including VKPM B-3386, MDC9632, and MDC9633 could not suppress the growth of any tested mold; however, the supplementation of ions combination revealed their antifungal effect against all kinds of molds. The finding of substantial stimulation of the most LAB associations antifungal effect by metal ions can be basis for creation of new effective antifungal preparations by the supplementation of ions combined mixture.

Antifungal and antibacterial effects of newly created lactic acid bacteria associations depending on cultivation media and duration of cultivation.

BACKGROUND: The newly created associations of lactic acid bacteria (LAB) isolated from Armenian dairy products (yoghurt, sour cream and different varieties of cheese), as well as from the gastrointestinal tract of honeybees were screened according to their antifungal and antibacterial activity. RESULTS: LAB strains were mixed at equal proportions (1:1) according to mathematical planning of experiments. Antifungal and antibacterial effects of different combinations (associations) were determined in different media, employing well-diffusion and total diffusion into agar methods. A number of fungal and bacterial test-organisms, including pathogenic ones, were used. Pure LAB strain cultures were used as a control. The antifungal effect of the most active strain Lactobacillus rhamnosus MDC 9661 in the associations with other LAB strains was partly decreased. At the same time, some mixed LAB cultures in DeMan, Rogosa and Sharpe (MRS) medium demonstrated significant antibacterial activity against wide spectra of test-organisms only in the case of simultaneous cultivation of LAB strains. On the other hand, in the case of different LAB strains cultivated in MRS with 24-h time break between mix formations by different strains, no inhibitory activity was revealed. But the inhibitory effect of many LAB associations against test-organisms was significantly increased in the case of separated cultivation in milk. CONCLUSION: The inhibitory effect of mixed LAB associations showed stronger dependence on the cultivation media and on the duration of cultivation with respect to each other. The co-cultivation of some strains, like L. rhamnosus MDC 9661, could lead to changed antagonistic activity. Consequently, the results are significant for creation and further investigation of LAB associations, as effective probiotics, and for their probable application in the production of antimicrobial preparations.

Growth of the facultative chemolithoautotroph Ralstonia eutropha on organic waste materials: growth characteristics, redox regulation and hydrogenase activity.

BACKGROUND: The chemolithoautotrophic ß-proteobacterium Ralstonia eutropha H16 (Cupriavidus necator) is one of the most studied model organisms for growth on H2 and CO2. R. eutropha H16 is also a biologically significant bacterium capable of synthesizing O2-tolerant [NiFe]-hydrogenases (Hyds), which can be used as anode biocatalysts in enzyme fuel cells. For heterotrophic growth of R. eutropha, various sources of organic carbon and energy can be used. RESULTS: Growth, bioenergetic properties, and oxidation-reduction potential (ORP) kinetics were investigated during cultivation of R. eutropha H16 on fructose and glycerol or lignocellulose-containing brewery spent grain hydrolysate (BSGH). BSGH was used as carbon and energy source by R. eutropha H16, and the activities of the membrane-bound hydrogenase (MBH) and cytoplasmic, soluble hydrogenase (SH) were measured in different growth phases. Growth of R. eutropha H16 on optimized BSGH medium yielded ~ 0.7 g cell dry weight L-1 with 3.50 ± 0.02 (SH) and 2.3 ± 0.03 (MBH) U (mg protein)-1 activities. Upon growth on fructose and glycerol, a pH drop from 7.0 to 6.7 and a concomitant decrease of ORP was observed. During growth on BSGH, in contrast, the pH and ORP stayed constant. The growth rate was slightly stimulated through addition of 1 mM K3[Fe(CN)6], whereas temporarily reduced growth was observed upon addition of 3 mM dithiothreitol. The overall and N,N'-dicyclohexylcarbodiimide-sensitive ATPase activities of membrane vesicles were ~ 4- and ~ 2.5-fold lower, respectively, upon growth on fructose and glycerol (FGN) compared with only fructose utilization (FN). Compared to FN, ORP was lower upon bacterial growth on FGN, GFN, and BSGH. CONCLUSIONS: Our results suggest that reductive conditions and low ATPase activity might be signals for energy depletion, which, in turn, leads to increased hydrogenase biosynthesis to overcome this unfavorable situation. Addition of fructose or microelements have no, or a negative, influence on hydrogenase activity. Organic wastes (glycerol, BSGH) are promising carbon and energy sources for the formation of biomass harboring significant amounts of the biotechnologically relevant hydrogenases MBH and SH. The results are valuable for using microbial cells as producers of hydrogenase enzymes as catalysts in enzymatic fuel cells.

The potential therapeutic effect of NG-hydroxy-nor-L-arginine in 7,12-dimethylbenz(a)anthracene-induced breast cancer in rats.

Advances in our understanding of the metabolism and molecular functions of arginine and their alterations in cancer have led to resurgence in the interest of targeting arginine catabolism as an anticancer strategy. Therefore, arginase inhibitors have been proposed as a way to treat cancer. In this study, the anti-tumor potential of the arginase inhibition by NG-hydroxy-nor-L-arginine (nor-NOHA) (3 mg/kg/day, i.p.), administered for 5 weeks (parallel tumors development, every 3th day) against 7,12-dimethylbenz(a)anthracene (DMBA)-induced mammary carcinogenesis in rats has been investigated. Treatment by nor-NOHA has obvious inhibition effects on development of carcinogenesis in rats was shown. That was seen in downregulation of rats' tumors size and number, mortality rate, in stopped alteration of tissue histopathology, in decrease of polyamines, NO and MDA (malondialdeide) concentrations (in blood). Results have shown arginase and NO-synthase can cooperate to restrain quantities of polyamines and NO for cancer progression. The results obtained can serve as a base to use this model for determination of productive, noncytotoxic antitumor and immune modulating concentration of anticancer agents. Perspectives of targeting arginase and NOS in cancer management can ground application in clinical medicine.

Antibacterial effects of iron oxide (Fe3O4) nanoparticles: distinguishing concentration-dependent effects with different bacterial cells growth and membrane-associated mechanisms.

Nowadays, the influence of nanoparticles (NPs) on microorganisms attracts a great deal of attention as an alternative to antibiotics. Iron oxide (Fe3O4) NPs' effects on Gram-negative Escherichia coli BW 25113 and Gram-positive Enterococcus hirae ATCC 9790 growth and membrane-associated mechanisms have been investigated in this study. Growth specific rate of E. coli was decreased, indicating the bactericidal effect of Fe3O4 NPs. This inhibitory effect of NPs had a concentration-dependent manner. The reactive oxygen species together with superoxide radicals and singlet oxygen formed by Fe3O4 NPs could be the inhibition cause. Fe3O4 NPs showed opposite effects on E. hirae: the growth stimulation or inhibition was observed depending on NPs concentration used. Addition of NPs altered redox potential kinetics and inhibited H2 yield in E. coli; no change in intracellular pH was determined. Fe3O4 NPs decreased H+-fluxes through bacterial membrane more in E. coli than in E. hirae even in the presence of DCCD and increased ATPase activity more in E. hirae than in E. coli. Our results showed that the Fe3O4 NPs demonstrate differentiating effects on Gram-negative and Gram-positive bacteria likely due to the differences in bacterial cell wall structure and metabolic peculiarities. Fe3O4 NPs of different concentrations have no hemolytic (cytotoxic) activity against erythrocytes. Therefore, they can be proposed as antibacterial agents in biomedicine, biotechnology, and pharmaceutics.

Insights into the Bacterial Diversity of the Acidic Akhtala Mine Tailing in Armenia Using Molecular Approaches.

The impact of the heavy metal contamination and acidity on the bacterial community was studied in samples collected from the Akhtala copper mine tailing using molecular approaches. The bacterial community structure analysis by PCR-DGGE fingerprinting revealed an abundance of Firmicutes, Acidobacteria, and Proteobacteria in different layers of the Akhtala tailing. 454 pyrotag sequence analyses revealed that a significant part of the sequences originated from Proteobacteria (49%) and Bacteroidetes (43%). Bacterial taxa are distributed also in phyla Saccharibacteria (2%), Verrucomicrobia (1.5%), Gammatimonadetes (1%), and some minor additional bacterial groups. The main primary producers in the Akhtala tailing appear to be obligate autotrophic Thiobacillus and Sulfuritalea species. Representatives of Lutibacter and Lysobacter genera are the most abundant acid-tolerant heterotrophs in the studied tailing. The presence of a large number of yet-uncultivated species emphasizes the importance of the future exploration of the tailing as an important source of novel bacteria.

Antibacterial activities of transient metals nanoparticles and membranous mechanisms of action.

Various transient metal and metal oxide nanoparticles (NPs) have shown pronounced biological activity, including antibacterial action against different Gram-negative and Gram-positive bacteria including pathogens and drug-resistant ones. Thus, NPs can be applied in nanotechnology for controlling bacterial growth as well as in biomedicine for the treatment of various diseases. However, the mechanisms of these effects are not clear yet. This review is focused on the antibacterial effects of transient metal NPs, especially iron oxide (Fe3O4) and Ag NPs on Escherichia coli wild type and antibiotic-resistant strains. Ag NPs show more pronounced bactericidal effect than Fe3O4 NPs. Moreover, Ag NPs display more expressed antibacterial effect at low concentrations. Interestingly, kanamycin-resistant strain is more susceptible to Fe3O4 NPs than wild type strain. In order to explain the possible mechanisms of NP effects, in addition to the production of reactive oxygen species causing damage in cells, particularly, their membranes, the changes in the membrane-associated H+-translocating FOF1-ATPase activity, H+-fluxes through the bacterial membrane, redox potential and hydrogen yield by membrane-associated enzymes-hydrogenases, are discussed. We observed from the results that FOF1-ATPase could be a main target for NPs. A scheme of possible action mechanism is proposed.

Understanding the Role of Escherichia coli Hydrogenases and Formate Dehydrogenases in the FO F1 -ATPase Activity during the Mixed Acid Fermentation of Mixture of Carbon Sources.

During fermentation Escherichia coli produces di-hydrogen (H2 ) via reversible membrane-bound [Ni-Fe]-hydrogenases (Hyd). This study describes the total and N,N'-dicyclohexylcarbodiimide (DCCD) inhibited ATPase activity and H2 production at various pHs in E. coli wild type and mutants encoding Hyd enzymes and formate dehydrogenases (FDH) on fermentation of glucose, glycerol, and formate. The highest total ATPase activity was detected at pH 7.5 in hyaB hybC selC (lacking large subunits of Hyd-1 and Hyd-2 and FDH, respectively) triple mutant. This ATPase activity was mainly due to the proton-translocating ATPase but in FDH mutant the DCCD inhibition was less compared to wild type. Potassium ions stimulated total ATPase activity at pH 5.5 ~50% and ~35% in wild type and hypF (lacking all Hyd enzymes) mutant, respectively. Moreover, K+ also stimulated DCCD inhibited ATPase activity ~1.7-fold-2-fold in strains where FDH was absent only at pH 5.5. DCCD inhibited H2 production only at pH 5.5 in all assays. Taken together it is suggested that at low pH, FDH, and Hyd enzymes are linked with the FO F1 -ATPase for regulating and maintaining the cytoplasmatic pH and thus proton motive force generation. FDH and Hyd enzymes have impact on the FO F1 -ATPase activity depending on external pH and potassium ions. © 2018 IUBMB Life, 70(10):1040-1047, 2018.

Identification and sequence analyses of novel lipase encoding novel thermophillic bacilli isolated from Armenian geothermal springs.

BACKGROUND: Among the huge diversity of thermophilic bacteria mainly bacilli have been reported as active thermostable lipase producers. Geothermal springs serve as the main source for isolation of thermostable lipase producing bacilli. Thermostable lipolytic enzymes, functioning in the harsh conditions, have promising applications in processing of organic chemicals, detergent formulation, synthesis of biosurfactants, pharmaceutical processing etc. RESULTS: In order to study the distribution of lipase-producing thermophilic bacilli and their specific lipase protein primary structures, three lipase producers from different genera were isolated from mesothermal (27.5-70 °C) springs distributed on the territory of Armenia and Nagorno Karabakh. Based on phenotypic characteristics and 16S rRNA gene sequencing the isolates were identified as Geobacillus sp., Bacillus licheniformis and Anoxibacillus flavithermus strains. The lipase genes of isolates were sequenced by using initially designed primer sets. Multiple alignments generated from primary structures of the lipase proteins and annotated lipase protein sequences, conserved regions analysis and amino acid composition have illustrated the similarity (98-99%) of the lipases with true lipases (family I) and GDSL esterase family (family II). A conserved sequence block that determines the thermostability has been identified in the multiple alignments of the lipase proteins. CONCLUSIONS: The results are spreading light on the lipase producing bacilli distribution in geothermal springs in Armenia and Nagorno Karabakh. Newly isolated bacilli strains could be prospective source for thermostable lipases and their genes.

The Effects of Different Carbon Sources on the Antifungal Activity by Lactic Acid Bacteria.

Lactobacillus rhamnosus R-2002 strain isolated from Armenian pickled cheese sample has a fungicide effect, which was carried out by the first-time designed method. The fungicide effect of LAB strain was expressed after 5 days of co-cultivation with Mucor plumbeus and 6 days of co-cultivation with Penicillium aurantioviolaceum. The growth as well as the antifungal activity of L. rhamnosus R-2002 was shown to directly depend on the nature of the medium's carbon source. The antifungal activity of this strain against both moulds was inhibited when the combination of 10 g l-1 glucose and 10 ml l-1 ethanol in the medium was used as a carbon source. The results could be applied in food industry, medicine, and veterinary.

Antimicrobial activity of some plant materials used in Armenian traditional medicine.

BACKGROUND: Antibiotic resistance has become one of the major problems facing humanity. The need for new antimicrobials has been increased dramatically. Plants are considered as one of the most promising sources for new antimicrobials discovery. Despite relatively small area, Armenia has large diversity of flora with many endemic species. In Armenian folk medicine plant materials have been used to treat various microbial diseases since ancient times. The goal of our research was to evaluate antimicrobial efficiency of different parts of five wild plants species which are commonly used in Armenian traditional medicine. METHODS: Plant crude extracts were obtained with maceration technique using five solvents separately: distilled water, methanol, chloroform, acetone, and hexane. Agar well diffusion assay was used for initial evaluation of antimicrobial properties of plant materials against five bacterial and two yeast strains. Minimum inhibitory concentrations of the most active plant parts were determined by broth microdilution method. RESULTS: Crude extracts of all five tested plants expressed antimicrobial activity against at least four test strains at 500 µg ml-1 concentration. Minimum inhibitory and bactericidal/fungicidal concentrations of selected plant parts were determined. Crude acetone and hexane extracts of Hypericum alpestre and acetone extract of Sanguisorba officinalis inhibited the growth of P. aeruginosa even at 64 µg ml-1 concentration. Chloroform and acetone extracts of Sanguisorba officinalis exhibited cidal activity against P. aeruginosa till 256 µg ml-1. Acetone was the most effective solvent for solubilizing antimicrobial compounds for almost all tested plant materials. CONCLUSIONS: Thus, antimicrobial activity of some medicinal plants used in Armenian traditional medicine was evaluated. Some of the plants had rather low minimum bacteriostatic/bactericidal concentrations and therefore they have prospective for further more inclusive studies.

Chemical composition and some biological activities of the essential oils from basil Ocimum different cultivars.

BACKGROUND: The plants belonging to the Ocimum genus of the Lamiaceae family are considered to be a rich source of essential oils which have expressed biological activity and use in different area of human activity. There is a great variety of chemotypes within the same basil species. Essential oils from three different cultivars of basil, O. basilicum var. purpureum, O. basilicum var. thyrsiflora, and O. citriodorum Vis. were the subjects of our investigations. METHODS: The oils were obtained by steam distillation in a Clevenger-type apparatus. The gas chromatography mass selective analysis was used to determine their chemical composition. The antioxidant activities of these essential oils were measured using 1,1-diphenyl-2-picrylhydrazyl assays; the tyrosinase inhibition abilities of the given group of oils were also assessed spectophotometrically, and the antimicrobial activity of the essential oils was determined by the agar diffusion method, minimal inhibitory concentrations were expressed. RESULTS: According to the results, the qualitative and quantitative composition of essential oils was quite different: O. basilicum var. purpureum essential oil contained 57.3% methyl-chavicol (estragol); O. basilicum var. thyrsiflora oil had 68.0% linalool. The main constituents of O. citriodorum oil were nerol (23.0%) and citral (20.7%). The highest antioxidant activity was demonstrated by O. basilicum var. thyrsiflora essential oil. This oil has also exhibited the highest tyrosinase inhibition level, whereas the oil from O. citriodorum cultivar demonstrated the highest antimicrobial activity. CONCLUSIONS: The results obtained indicate that these essential oils have antioxidant, antibacterial and antifungal activity and can be used as natural antioxidant and antimicrobial agents in medicine, food industry and cosmetics.

Millimeter waves or extremely high frequency electromagnetic fields in the environment: what are their effects on bacteria?

Millimeter waves (MMW) or electromagnetic fields of extremely high frequencies at low intensity is a new environmental factor, the level of which is increased as technology advance. It is of interest that bacteria and other cells might communicate with each other by electromagnetic field of sub-extremely high frequency range. These MMW affected Escherichia coli and many other bacteria, mainly depressing their growth and changing properties and activity. These effects were non-thermal and depended on different factors. The significant cellular targets for MMW effects could be water, cell plasma membrane, and genome. The model for the MMW interaction with bacteria is suggested; a role of the membrane-associated proton FOF1-ATPase, key enzyme of bioenergetic relevance, is proposed. The consequences of MMW interaction with bacteria are the changes in their sensitivity to different biologically active chemicals, including antibiotics. Novel data on MMW effects on bacteria and their sensitivity to different antibiotics are presented and discussed; the combined action of MMW and antibiotics resulted with more strong effects. These effects are of significance for understanding changed metabolic pathways and distinguish role of bacteria in environment; they might be leading to antibiotic resistance in bacteria. The effects might have applications in the development of technique, therapeutic practices, and food protection technology.

Mechanisms for hydrogen production by different bacteria during mixed-acid and photo-fermentation and perspectives of hydrogen production biotechnology.

H2 has a great potential as an ecologically-clean, renewable and capable fuel. It can be mainly produced via hydrogenases (Hyd) by different bacteria, especially Escherichia coli and Rhodobacter sphaeroides. The operation direction and activity of multiple Hyd enzymes in E. coli during mixed-acid fermentation might determine H2 production; some metabolic cross-talk between Hyd enzymes is proposed. Manipulating the activity of different Hyd enzymes is an effective way to enhance H2 production by E. coli in biotechnology. Moreover, a novel approach would be the use of glycerol as feedstock in fermentation processes leading to H2 production. Mixed carbon (sugar and glycerol) utilization studies enlarge the kind of organic wastes used in biotechnology. During photo-fermentation under limited nitrogen conditions, H2 production by Rh. sphaeroides is observed when carbon and nitrogen sources are supplemented. The relationship of H2 production with H(+) transport across the membrane and membrane-associated ATPase activity is shown. On the other hand, combination of carbon sources (succinate, malate) with different nitrogen sources (yeast extract, glutamate, glycine) as well as different metal (Fe, Ni, Mg) ions might regulate H2 production. All these can enhance H2 production yield by Rh. sphaeroides in biotechnology Finally, two of these bacteria might be combined to develop and consequently to optimize two stages of H2 production biotechnology with high efficiency transformation of different organic sources.