Gold nanoparticles activate hydrogenase synthesis and improve heterotrophic growth of Ralstonia eutropha H16.

Ralstonia eutropha is a facultative chemolithoautotrophic aerobic bacterium that grows using organic substrates or H2 and CO2. Hydrogenases (Hyds) are synthesized under lithoautotrophic, or energy-limited heterotrophic conditions and are used in enzyme fuel cells (EFC) as anodic catalysts. The effects of chemically synthesized gold nanoparticles (Au-NPs) on R. eutropha H16 growth, oxidation-reduction potential (ORP) kinetics, and H2-oxidizing Hyd activity were investigated in this study. Atomic force microscopy showed that thin, plate-shaped Au-NPs were in the nanoscale range with an average size of 5.68 nm. Compared with growth in medium without Au-NPs (control), the presence of Au-NPs stimulated growth, and resulted in a decrease in ORP to negative values. H2-oxidizing activity was not detected in the absence of Au-NPs, but activity was significantly induced (12 U/g CDW) after 24 h of growth with 18 ng/ml, increasing a further 4-fold after 72 h of growth. The results demonstrate that Au-NPs primarily influence the membrane-bound Hyd. In contrast to R. eutropha, Au-NPs had a negligible or negative effect on the growth, Hyd activity, and H2 production of Escherichia coli. The findings of this study offer new perspectives for the production of oxygen-tolerant Hyds and the development of EFCs.

Ribes nigrum L. Extract-Mediated Green Synthesis and Antibacterial Action Mechanisms of Silver Nanoparticles.

Silver nanoparticles (Ag NPs) represent one of the most widely employed metal-based engineered nanomaterials with a broad range of applications in different areas of science. Plant extracts (PEs) serve as green reducing and coating agents and can be exploited for the generation of Ag NPs. In this study, the phytochemical composition of ethanolic extract of black currant (Ribes nigrum) leaves was determined. The main components of extract include quercetin rutinoside, quercetin hexoside, quercetin glucuronide, quercetin malonylglucoside and quercitrin. The extract was subsequently employed for the green synthesis of Ag NPs. Consequently, R. nigrum leaf extract and Ag NPs were evaluated for potential antibacterial activities against Gram-negative bacteria (Escherichia coli ATCC 25922 and kanamycin-resistant E. coli pARG-25 strains). Intriguingly, the plant extract did not show any antibacterial effect, whilst Ag NPs demonstrated significant activity against tested bacteria. Biogenic Ag NPs affect the ATPase activity and energy-dependent H+-fluxes in both strains of E. coli, even in the presence of N,N'-dicyclohexylcarbodiimide (DCCD). Thus, the antibacterial activity of the investigated Ag NPs can be explained by their impact on the membrane-associated properties of bacteria.

Biosynthesis of silver nanoparticles using extracts of Stevia rebaudiana and evaluation of antibacterial activity.

The present study reveals a simple, non-toxic and eco-friendly method for the "green" synthesis of Ag-NPs using hydroponic and soil medicinal plant Stevia rebaudiana extracts, the characterization of biosynthesized nanoparticles, as well as the evaluation of their antibacterial activity. Transmission electronic microscopy (TEM) and Dynamic Light Scattering (DLS) analysis confirmed that biosynthesized Ag-NPs are in the nano-size range (50-100 nm) and have irregular morphology. Biogenic NPs demonstrate antibacterial activity against Escherichia coli BW 25,113, Enterococcus hirae ATCC 9790, and Staphylococcus aureus MDC 5233. The results showed a more pronounced antibacterial effect on E. coli growth rate, in comparison with Gram-positive bacteria, which is linked to the differences in the structure of bacterial cell wall. Moreover, the Ag-NPs not only suppressed the growth of bacteria but also changed the energy-dependent H+-fluxes across the bacterial membrane. The change of H+-fluxes in presence of H+-translocating systems inhibitor, N,N'-dicyclohexylcarbodiimide (DCCD), proves the effect of Ag-NPs on the structure and permeability of the bacterial membrane. Overall, our findings indicate that the Ag-NPs synthesized by medicinal plant Stevia extracts may be an excellent candidate as an alternative to antibiotics against the tested bacteria.

Antibacterial activity of royal jelly-mediated green synthesized silver nanoparticles.

The application of green synthesis in nanotechnology is growing day by day. It's a safe and eco-friendly alternative to conventional methods. The current research aimed to study raw royal jelly's potential in the green synthesis of silver nanoparticles and their antibacterial activity. Royal jelly served as a reducing and oxidizing agent in the green synthesis technology of colloidal silver nanoparticles. The UV-Vis maximum absorption at ~ 430 nm and fluorescence emission peaks at ~ 487 nm confirmed the presence of Ag NPs. Morphology and structural properties of Ag NPs and the effect of ultrasound studies revealed: (i) the formation of polydispersed and spherical particles with different sizes; (ii) size reduction and homogeneity increase by ultrasound treatment. Antibacterial activity of different concentrations of green synthesized Ag NPs has been assessed on Gram-negative S. typhimurium and Gram-positive S. aureus, revealing higher sensitivity on Gram-negative bacteria.

Systemic Sclerosis: Current State and Survival After Lung Transplantation.

Systemic sclerosis (SSc) is an autoimmune disorder characterized by the involvement of skin and internal organs. With the introduction of angiotensin-converting enzyme inhibitors (ACEIs), scleroderma renal crisis (SRC) is no longer considered a leading cause of death in affected patients. In fact, pulmonary manifestations [interstitial lung disease (ILD) and pulmonary arterial hypertension (PAH)] are currently the major cause of death in patients with SSc. Historically, many centers have been reluctant to offer lung transplantation to patients with SSc due to multiple extrapulmonary manifestations and the assumption of poor post-transplant survival. The purpose of this review is to highlight the recent advances in the evaluation and management of patients with pulmonary manifestations of SSc. We also engage in a systematic literature review to assess all the available data on the survival of patients with SSc after lung transplantation.

Behavioral Deficits and Brain α-Synuclein and Phosphorylated Serine-129 α-Synuclein in Male and Female Mice Overexpressing Human α-Synuclein.

BACKGROUND: Alpha-synuclein (α-syn) is involved in pathology of Parkinson's disease, and 90% of α-syn in Lewy bodies is phosphorylated at serine 129 (pS129 α-syn). OBJECTIVE: To assess behavior impairments and brain levels of α-syn and pS129 α-syn in mice overexpressing human α-syn under Thy1 promoter (Thy1-α-syn) and wild type (wt) littermates. METHODS: Motor and non-motor behaviors were monitored, brain human α-syn levels measured by ELISA, and α-syn and pS129 α-syn mapped by immunohistochemistry. RESULTS: Male and female wt littermates did not show differences in the behavioral tests. Male Thy1-α-syn mice displayed more severe impairments than female counterparts in cotton nesting, pole tests, adhesive removal, finding buried food, and marble burying. Concentrations of human α-syn in the olfactory regions, cortex, nigrostriatal system, and dorsal medulla were significantly increased in Thy1-α-syn mice, higher in males than females. Immunoreactivity of α-syn was not simply increased in Thy1-α-syn mice but had altered localization in somas and fibers in a few brain areas. Abundant pS129 α-syn existed in many brain areas of Thy1-α-syn mice, while there was none or only a small amount in a few brain regions of wt mice. The substantia nigra, olfactory regions, amygdala, lateral parabrachial nucleus, and dorsal vagal complex displayed different distribution patterns between wt and transgenic mice, but not between sexes. CONCLUSION: The severer abnormal behaviors in male than female Thy1-α-syn mice may be related to higher brain levels of human α-syn, in the absence of sex differences in the altered brain immunoreactivity patterns of α-syn and pS129 α-syn.

Growth properties and hydrogen yield in green microalga Parachlorella kessleri: Effects of low-intensity electromagnetic irradiation at the frequencies of 51.8 GHz and 53.0 GHz.

The current research reports the effects of low-intensity extremely high frequency electromagnetic irradiation (EMI) of 51.8 GHz and 53.0 GHz on green microalga Parachlorella kessleri RA-002 isolated in Armenia. EMI demonstrated different effects on the growth properties of microalgae under various conditions. Under aerobic conditions a positive effect of EMI on the growth rate of P. kessleri and the content of photosynthetic pigments were observed. The data obtained indicates a significant role of O2, since the enhancing effect of EMI was determined only under aerobic conditions. Meanwhile under anaerobic conditions EMI with both frequencies caused inhibition of algal growth and a decrease in the amount of photosynthetic pigments. EMI also inhibited the yield of H2 production in P. kessleri, which was partially restored after 5-day cultivation due to the existence of protective mechanisms in this alga. The results might indicate membrane-bound mechanisms of EMI action on algae, which can be associated with the effects on photosynthetic pigments and membrane-associated enzymes responsible for H2 production. The results are useful for the development of algae biotechnology and the possibility of using EMI as a factor which regulates the production of biomass and biohydrogen by green microalgae.

Comparable antibacterial effects and action mechanisms of silver and iron oxide nanoparticles on Escherichia coli and Salmonella typhimurium.

The current research reports the antibacterial effects of silver (Ag) and citric acid coated iron oxide (Fe3O4) NPs on Escherichia coli wild type and kanamycin-resistant strains, as well as on Salmonella typhimurium MDC1759. NPs demonstrated significant antibacterial activity against these bacteria, but antibacterial effect of Ag NPs is more pronounced at low concentrations. Ag NPs inhibited 60-90% of S. typhimurium and drug-resistant E. coli. The latter is more sensitive to Fe3O4 NPs than wild type strain: the number of bacterial colonies is decreased ~ 4-fold. To explain possible mechanisms of NPs action, H+-fluxes through the bacterial membrane and the H+-translocating FOF1-ATPase activity of bacterial membrane vesicles were studied. N,N'-Dicyclohexylcarbodiimide (DCCD)-sensitive ATPase activity was increased up to ~ 1.5-fold in the presence of Fe3O4 NPs. ATPase activity was not detected by Ag NPs even in the presence of DCCD, which confirms the bactericidal effect of these NPs. The H+-fluxes were changed by NPs and by addition of DCCD. H2 yield was inhibited by NPs; the inhibition by Ag NPs is stronger than by Fe3O4 NPs. NPs showed antibacterial effect in bacteria studied in concentration-dependent manner by changing in membrane permeability and membrane-bound enzyme activity. The FOF1-ATPase is suggested might be a target for NPs.

Silver ion bioreduction in nanoparticles using Artemisia annua L. extract: characterization and application as antibacterial agents.

The biological synthesis of metal nanoparticles using plant extracts with defined size and morphology is a simple, nontoxic and environmentally friendly method. The present study focused on the synthesis of silver nanoparticles (Ag NPs) by Artemisia annua L. extract as reducing and stabilising agent. The Ag NPs function, as antibacterial agents, is with that they are further used in human therapy. The effects of pH and temperature on the synthesis of NPs were characterized by UV-absorption spectroscopy and shown by surface plasmon resonance (SPR) band at 410 nm. NPs' size and morphology were measured by transmission electron microscopy (TEM) and dynamic light scattering (DLS). TEM images showed that Ag NPs were in a nano-sized range (20-90 nm) and had spherical shape. Our findings demonstrated that lower concentration (100 µg mL-1) of the biogenic Ag NPs exhibited antibacterial activity against Gram-negative Escherichia coli BW 25113 and Gram-positive Enterococcus hirae ATCC 9790.

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.

Regulation of biohydrogen production by protonophores in novel green microalgae Parachlorella kessleri.

The green microalgae Parachlorella kessleri RA-002 isolated in Armenia can produce biohydrogen (H2) during oxygenic photosynthesis. Addition of protonophores, carbonyl cyanide m-chlorophenylhydrazone (CCCP) and 2,4-dinitrophenol (DNF) enhances H2 yield in P. kessleri. The maximal H2 yield of ~2.20 and 2.08 mmol L-1 was obtained in the presence of 15 µM CCCP and 50 µM DNF, respectively. During dark conditions H2 production by P. kessleri was not observed even in the presence of protonophores, indicating that H2 formation in these algae was mediated by light conditions. The enhancing effect of protonophores can be coupled with dissipation of proton motive force across thylakoid membrane in P. kessleri, facilitating the availability of protons and electrons to [Fe-Fe]-hydrogenase, which led to formation of H2. At the same time H2 production was not observed in the presence of diuron (3-(3,4-dichlorophenyl)-1,1-dimethylurea), a specific inhibitor of PS II. Moreover, diuron inhibits H2 yield in P. kessleri in the presence of protonophores. The inhibitory effect of diuron coupled with suppression of electron transfer from PS II. The results showed that in these algae operates PS II-dependent pathway of H2 generation. This study is important for understanding of the mechanisms of H2 production by green microalgae P. kessleri and developing of its biotechnology.

Multiple Beneficial Effects of Ghrelin Agonist, HM01 on Homeostasis Alterations in 6-Hydroxydopamine Model of Parkinson's Disease in Male Rats.

Background and objective: Developing therapy for non-motor symptoms of Parkinson's disease (PD) is important for improving patients' quality of life. Previously, we reported that the ghrelin receptor agonist, HM01 normalized the decreased 4-h fecal output and levodopa-inhibited gastric emptying in 6-OHDA rats, and activated selective areas in brain and spinal cord. In this study, we evaluated whether chronic HM01 treatment influences motor functions and/or has beneficial effects on non-motor symptoms including alterations of body weight and composition, defecation, feeding and water intake in 6-OHDA rats. Methods: Male rats were microinjected unilaterally into the medial forebrain bundle with either vehicle or 6-OHDA. Three weeks later, we assessed basal body weight, and 24-h fecal output (pellets, weight, dry weight and water content), water intake and food intake (ingested and spillage). Then, HM01 (3 mg/kg) or vehicle was given per gavage daily for 10-12 days and the same parameters were re-assessed daily. Motor behavior (stepping and rotations tests), body composition were monitored before and after the HM01 treatment. Results: 6-OHDA rats showed motor deficits in rotation test induced by apomorphine and stepping test. They also displayed a significant reduction in body weight, water consumption, fecal weight and water content and an increase in food spillage compared to vehicle microinjected rats. Daily oral treatment of HM01 did not modify motor alterations compared to vehicle but significantly increased the body weight, fat mass, and 24-h fecal weight, fecal water content, food and water intake in 6-OHDA rats, while HM01 had no significant effect in vehicle microinjected rats. Fecal weight and water content were both correlated with water intake, but not with food intake. Fat mass, but not body weight, was correlated with food intake. HM01 effects were significant after 24 h and remained similar during the treatment. Conclusions: Chronic treatment with ghrelin agonist, HM01 improved several non-motor symptoms in the rat PD model induced by 6-OHDA lesion including the decrease in body weight, water consumption, fecal weight and water content, and increased food intake while not improving the motor deficits. These findings provide pre-clinical evidence of potential benefits of ghrelin agonists to alleviate non-motor symptoms in PD patients.

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.

Characterization of light-dependent hydrogen production by new green microalga Parachlorella kessleri in various conditions.

Nowadays, hydrogen (H2) production by green microalgae seems to be a very perspective, as stocks of water and solar energy are practically inexhaustible and renewable. The aim of this study was the optimization of conditions (organic carbon sources and lighting regime), which can provide light-dependent H2 production by green microalga Parachlorella kessleri RA-002 newly isolated in Armenia. The results indicated that carbon sources and lighting regimes affected H2 production. In the presence of used carbon sources H2 production was observed, but the highest yield of H2 was obtained in the presence of acetate. It was 2-fold higher than the H2 yield determined in the presence of glucose. The increase of H2 production might be connected with the stimulation of H2-producing enzyme - [Fe]-hydrogenase synthesis. The data obtained show that acetate can be used as an effective carbon source in H2 production. H2 production by microalga (in the presence of acetate and glucose) was enhanced by 1.5-2.5-fold in comparison with continuously illuminated algal cells, when P. kessleri was illuminated during 24h, and then was moved in the darkness. H2 yield increase is possible due to hydrogenase activation and the creation of anaerobic conditions. This study was significant to find out available effective substrates and optimal lighting regime to provide with light-dependent H2 production by microalgae.

The Gastric and Intestinal Microbiome: Role of Proton Pump Inhibitors.

PURPOSE OF REVIEW: The discovery of Helicobacter pylori and other organisms colonizing the stomach and the intestines has shed some light on the importance of microbiome in maintaining overall health and developing pathological conditions when alterations in biodiversity are present. The gastric acidity plays a crucial role in filtering out bacteria and preventing development of enteric infections. In this article, we discuss the physiology of gastric acid secretion and bacterial contribution to the composition of gastric and intestinal barriers and review the current literature on the role of proton pump inhibitors (PPIs) in the microbial biodiversity of the gastrointestinal tract. RECENT FINDINGS: Culture-independent techniques, such as 16S rRNA sequencing, have revolutionized our understanding of the microbial biodiversity in the gastrointestinal tract. Luminal and mucosa-associated microbial populations are not identical. Streptococcus is overrepresented in the biopsies of patients with antral gastritis and may also be responsible for the development of peptic ulcer disease. The use of PPIs favors relative streptococcal abundance irrespective of H. pylori status and may explain the persistence of dyspeptic symptoms in patients on PPI therapy. Increased risk of enteric infections has also been seen in patients taking PPIs. The overuse of PPIs leads to significant shift of the gastrointestinal microbiome towards a less healthy state. With the advent of PPIs, many studies have demonstrated the significant changes in the microbial composition of both gastric and intestinal microbiota. Although they are considered relatively safe over-the-counter medications, PPIs in many cases are over- and even inappropriately used. Future studies assessing the safety of PPIs and their role in the development of microbiome changes should be encouraged.

Comparative effects of Ni(II) and Cu(II) ions and their combinations on redox potential and hydrogen photoproduction by Rhodobacter sphaeroides.

The aim of the present work was the study of comparative effects of Cu(II) and Ni(II) and their mixture on growth, redox potential, hydrogen (H2) yield and ATPase activity in phototrophic purple bacteria R. sphaeroides MDC6522 from Jermuk mineral spring in Armenia. It was ascertained, that Cu2+ and Ni2+ have different effects on bacterial specific growth rate: in the presence of 5µM Cu2+ growth rate was ~3.2-fold lower in comparison with control (no addition), and increased ~1.5-fold in medium with 5µM Ni2+. These changes may be resulted by action of the ions on redox potential (Eh). Low concentrations of Ni2+ had an enhancing effect on the Eh drop and H2 production. The increase of concentration from 1 to 5µM enhanced the stimulatory effect of Ni2+. H2 yield in R. sphaeroides (72h of growth) was enhanced ~3-fold with 5µM Ni2+, whereas in the presence of 5µM Cu2+ H2 yield was ~1.2 fold lower in comparison with control. Cu2++Ni2+ combinations effects were differed from the effect when ions used separately. When Cu2+ and Ni2+ were added together, the Ni2+ stimulatory effect disappeared, which indicated that heavy metal ions mixture may have different action mechanisms. Moreover, N,N'-dicyclohexylcarbodiimide-sensitive ATPase activity of R. sphaeroides membrane vesicles has been increased in the presence of both ions, but in the presence of Сu2+ the influence was feebly marked in comparison with Ni2+. The results suggest an interaction between these ions and the FOF1-ATPase. Thus, the results obtained point out discrimination between Cu2+ and Ni2+ and their combinations effects and reveal new regulatory pathways to enhance H2 yield in R. sphaeroides.

Biohydrogen production by purple non-sulfur bacteria Rhodobacter sphaeroides: Effect of low-intensity electromagnetic irradiation.

The present work was focused on the effects of low-intensity (the flux capacity was of 0.06mWcm(-2)) electromagnetic irradiation (EMI) of extremely high frequencies or millimeter waves on the growth and hydrogen (H2) photoproduction by purple non-sulfur bacteria Rhodobacter sphaeroides MDC6521 (from Armenian mineral springs). After exposure of R. sphaeroides, grown under anaerobic conditions upon illumination, to EMI (51.8GHz and 53.0GHz) for 15min an increase of specific growth rate by ~1.2-fold, in comparison with control (non-irradiated cells), was obtained. However, the effect of EMI depends on the duration of irradiation: the exposure elongation up to 60min caused the delay of the growth lag phase and the decrease specific growth rate by ~1.3-fold, indicating the bactericidal effect of EMI. H2 yield of the culture, irradiated by EMI for 15min, determined during 72h growth, was ~1.2-fold higher than H2 yield of control cells, whereas H2 production by cultures, irradiated by EMI for 60min was not observed during 72h growth. This difference in the effects of extremely high frequency EMI indicates a direct effect of radiation on the membrane transfer and the enzymes of these bacteria. Moreover, EMI increased DCCD-inhibited H(+) fluxes across the bacterial membrane and DCCD-sensitive ATPase activity of membrane vesicles, indicating that the proton FoF1-ATPase is presumably a basic target for extremely high frequency EMI related to H2 production by cultures.

Novel properties of photofermentative biohydrogen production by purple bacteria Rhodobacter sphaeroides: effects of protonophores and inhibitors of responsible enzymes.

BACKGROUND: Biohydrogen (H2) production by purple bacteria during photofermentation is a very promising way among biological H2 production methods. The effects of protonophores, carbonyl cyanide m-chlorophenylhydrazone (CCCP), 2,4-dinitrophenol (DNP), and inhibitors of enzymes, involved in H2 metabolism, metronidazole (Met), diphenyleneiodonium (DPI), and dimethylsulphoxide (DMSO) on H2 production by Rhodobacter sphaeroides MDC6522 isolated from Jermuk mineral springs in Armenia have been investigated in both nitrogen-limited and nitrogen-excess conditions. RESULTS: With the increase of inhibitors concentrations H2 yield gradually decreased. The complete inhibition of H2 production was observed in the presence of DPI and CCCP. DPI's solvent-DMSO in low concentration did not significantly affect H2 yield. N,N'-dicyclohexylcarbodiimide (DCCD)-inhibited the FOF1-ATPase activity of bacterial membrane vesicles was analyzed in the presence of inhibitors. Low concentrations of DPI and DMSO did not affect ATPase activity, whereas Met and CCCP stimulated enzyme activity. The effect of DNP was similar to CCCP. CONCLUSIONS AND SIGNIFICANCE: The results have shown the low concentration or concentration dependent effects of protonophores and nitrogenase and hydrogenase inhibitors on photofermentative H2 production by Rh. sphaeroides in nitrogen-limited and nitrogen-excess conditions. They would be significant to understand novel properties in relationship between nitrogenase, hydrogenase and the FOF1-ATPase in Rh. sphaeroides, and regulatory pathways of photofermentation. The inhibitors of nitrogenase and hydrogenase can be used in biotechnology for regulation of H2 production in different technology conditions and development of scale-up applications, for biomass and energy production using purple bacterial cells.

Relationship of proton motive force and the F(0)F (1)-ATPase with bio-hydrogen production activity of Rhodobacter sphaeroides: effects of diphenylene iodonium, hydrogenase inhibitor, and its solvent dimethylsulphoxide.

Rhodobacter sphaeroides MDC 6521 was able to produce bio-hydrogen (H(2)) in anaerobic conditions under illumination. In this study the effects of the hydrogenase inhibitor-diphenylene iodonium (Ph(2)I) and its solvent dimethylsulphoxide (DMSO) on growth characteristics and H(2) production by R. sphaeroides were investigated. The results point out the concentration dependent DMSO effect: in the presence of 10 mM DMSO H(2) yield was ~6 fold lower than that of the control. The bacterium was unable to produce H(2) in the presence of Ph(2)I. In order to examine the mediatory role of proton motive force (∆p) or the F(0)F(1)-ATPase in H(2) production by R. sphaeroides, the effects of Ph(2)I and DMSO on ∆p and its components (membrane potential (∆ψ) and transmembrane pH gradient), and ATPase activity were determined. In these conditions ∆ψ was of -98 mV and the reversed ∆pH was +30 mV, resulting in ∆p of -68 mV. Ph(2)I decreased ∆ψ in concentrations of 20 µM and higher; lower concentrations of Ph(2)I as DMSO had no valuable effect on ∆ψ. The R. sphaeroides membrane vesicles demonstrated significant ATPase activity sensitive to N,N'-dicyclohexylcarbodiimide. The 10-20 µM Ph(2)I did not affect the ATPase activity, whereas 40 µM Ph(2)I caused a marked inhibition (~2 fold) in ATPase activity. The obtained results provide novel evidence on the involvement of hydrogenase and the F(0)F(1)-ATPase in H(2) production by R. sphaeroides. Moreover, these data indicate the role of hydrogenase and the F(0)F(1)-ATPase in ∆p generation. In addition, DMSO might increase an interaction of nitrogenase with CO(2), decreasing nitrogenase activity and affecting H(2) production.