Bioconversion of synthesis gas to hydrogen using a light-dependent photosynthetic bacterium,Rhodospirillum rubrum

Biological hydrogen production from synthesis gas was carried out in batch culture. The phototrophic anaerobic bacterium, Rhodospirillum rubrum was used to oxidize CO and water to CO₂ and hydrogen. The bacteria were grown under anaerobic conditions in liquid medium; also acetate was used as carbon source in presence of synthesis gas. Biological hydrogen production was catalysed by R. rubrum via the water–gas shift reaction. A light-dependent cell growth modelled with a desired rate of hydrogen production and CO uptake was determined. The effect of light intensity on microbial cell growth was also studied at 500, 1,000 and 1,500 m.cd. A complete conversion of CO to hydrogen and maximum light efficiency were obtained with an acetate concentration of 1 g/l and light intensity of 500 m.cd. Utilization of the carbon monoxide from the gas phase was often considered as a mass transfer limited process, which needed to diffuse through the gas–liquid interface and then further diffuse into liquid medium prior to reaction. The results from this study showed that maximum cell propagation and hydrogen production were achieved with a limited light intensity of 1,000 m.cd. It was also found that high-light intensity may interfere with cell metabolism. In low-light intensity and substrate concentration, no inhibition was observed, however at extreme conditions, non-competitive inhibition was identified. The adverse effect of high-light intensity was shown at 5,000 m.cd, where the CO conversion drastically dropped to as low as 21%. Maximum CO conversion of 98% and maximum yield of 86% with an acetate concentration of 1.5 g/l and a light intensity of 1,000 m.cd were achieved.     

Hydrogen production by photoreactive nanoporous latex coatings of nongrowing Rhodopseudomonas palustris CGA009

Nonuniform light distribution is a fundamental limitation to biological hydrogen production by phototrophic bacteria. Numerous light distribution designs and culture conditions have been developed to reduce self-shading and nonuniform reactivity within bioreactors. In this study, highly concentrated (2.0 x 108 CFU/muL formulation) nongrowing Rhodopseudomonas palustris CGA009 were immobilized in thin, nanoporous, latex coatings. The coatings were used to study hydrogen production in an argon atmosphere as a function of coating composition, thickness, and light intensity. These coatings can be generated aerobically or anaerobically and are more reactive than an equivalent number of suspended or settled cells. Rhodopseudomonas palustris latex coatings remained active after hydrated storage for greater than 3 months in the dark and over 1 year when stored at -80 degrees C. The initial hydrogen production rate of the microphotobioreactors containing 6.25 cm2, 58.4 mum thick Rps. palustris latex coatings illuminated by 34.1 PAR mumol photons m-2 s-1 was 6.3 mmol H2 m-2 h-1 and had a final yield of 0.55 mol H2 m-2 in 120 h. A dispersible latex blend has been developed for direct comparison of the specific activity of settled, suspended, and immobilized Rps. palustris.     

Gaining electricity from in situ oxidation of hydrogen produced by fermentative cellulose degradation

AIM: To exploit the fermentative hydrogen generation and direct hydrogen oxidation for the generation of electric current from the degradation of cellulose. METHODS AND RESULTS: Utilizing the metabolic activity of the mesophilic anaerobe Clostridium cellulolyticum and the thermophilic Clostridium thermocellum we show that electricity generation is possible from cellulose fermentation. The current generation is based on an in situ oxidation of microbially synthesized hydrogen at platinum-poly(tetrafluoroaniline) (Pt-PTFA) composite electrodes. Current densities of 130 mA l(-1) (with 3 g cellulose per litre medium) were achieved in poised potential experiments under batch and semi-batch conditions. Conclusions: The presented results show that electricity generation is possible by the in situ oxidation of hydrogen, product of the anaerobic degradation of cellulose by cellulolytic bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: For the first time, it is shown that an insoluble complex carbohydrate like cellulose can be used for electricity generation in a microbial fuel cell. The concept represents a first step to the utilization of macromolecular biomass components for microbial electricity generation.     

Effect of ammonia on the growth of Bacillus species and some other bacteria

The tolerance of 26 Bacillus species isolated from alkaline fermented foods, five other bacilli and nine non spore-forming bacteria to alkaline pH and ammonia was determined. All grew at pH 7, 8 and 9 in the presence of 930 mmol l-1 NH4 + at pH 7.0, and in the presence of NH3 concentrations up to 5 mmol l-1 at pH 7.0 and 8.0. At higher NH3 concentrations, growth of some of the bacteria was inhibited and at 500 mmol l-1 only B. pasteurii and B. pumilus grew. Bacteria from alkaline food fermentations included strains relatively sensitive to NH3 (inhibited by 50 mmol l-1) and relatively tolerant strains (grew in the presence of 300 mmol l-1) and there was no evidence that they were more tolerant to NH3 than bacteria not associated with these fermentations.     

Effects of thioridazine on mechanical responses of human vas deferens induced by noradrenaline or potassium.

Thioridazine (0.1-10 mumol l-1) inhibited shortening of specimens of human vasa deferentia induced by noradrenaline (100 mumol l-1) or high extracellular potassium (136 mmol l-1). Thioridazine did not inhibit the lengthening response. In Ca(2+)-free media with EGTA (0.5 mmol l-1) similar results were obtained with responses to noradrenaline, but exposure to potassium elicited small contractions that were potentiated by thioridazine. Both shortening and lengthening responses to noradrenaline were antagonized by the alpha-adrenoceptor blockers prazosin (1-10 mumol l-1) and phentolamine (1-10 mumol l-1) and by the Ca2+ antagonists verapamil (10 mumol l-1) and diltiazem (10 mumol l-1). Responses to potassium were virtually abolished by the Ca2+ antagonists. These results show that thioridazine specifically inhibits longitudinal muscle of the human vas deferens and that its action cannot be entirely accounted for by a blockade of voltage-gated Ca2+ channels.     

Flocculation and cell surface characterization of Kloeckera apiculata from wine

AIMS: To characterize and analyze the flocculation phenomenon of Kloeckera apiculata mc1 from Argentinian wine to understand the cell-cell interaction pattern. METHODS AND RESULTS: Kloeckera apiculata mc1 possess intense cell-cell interactions in MYPG medium (0.5% malt extract, 1% yeast extract, 2% glucose, 2% peptone), pH 5.5 by shaking at 25 degrees C. Optimum flocculation is observed at pH 4.5 in the presence of 3 mmol l-1 Ca2+. The flocculation is induced by peptone and malt extract and not by yeast extract and is reversed by 50 mmol l-1 galactose or lactose. The flocculation is highly susceptible to pronase, chymotrypsine and proteases types IV and XXVII and is partially resistant to trypsin. The electronic microscopy shows that the cells are attached to each other along their sides by fine hair-like threads. CONCLUSIONS: The mechanism of flocculation of K. apiculata mc1 is mediated by protein-carbohydrate interaction, stabilized by Ca2+. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of selected pure yeast inocula of known ability is preferred to wine elaboration, therefore the indigenous flora must be avoided and the flocculation of K. apiculata could be an economic method to do it.     

Monitoring of potassium-stimulated catecholamine changes in striatal synaptosomal preparations and in corpus striatum of rats: a comparative voltammetric study.

Voltammetric techniques were used to compare the effects of K(+)-induced depolarization on catecholamine levels in in vitro synaptosomal preparations of the corpus striatum with those in the in vivo corpus striatum of anaesthetized animals. In vitro, the catechol-oxidation currents could be recorded only in dopamine-preloaded synaptosomes. In isolated synaptosomes prepared in the presence of elevated concentrations of Ca2+ (1 mmol.l-1) and Na+ (135 mmol.l-1), K(+)-induced depolarization had variable effects on catechol-oxidation current. The stimulatory effect of K(+)-induced depolarization (a short transient increase of catechol-oxidation current lasting for 30 s) could be observed after the addition of dopamine loaded synaptosomes in EGTA into the medium with elevated K+ concentration (90 mmol.l-1) and decreased concentrations of Na+ (75 mmol.l-1) and Ca2+ (0.75 mmol.l-1). These results suggest that experimental procedures and parameters of ionic composition of incubation media have to be carefully controlled, owing to an enhanced in vitro permeability of membranes of isolated synaptosomes for Ca2+ and Na+. In in vivo experiments, microinjection of KCl (3 microliters of 0.5 mol.l-1 KCl in 10 mmol.l-1 HEPES, pH 7.4) resulted in the appearance of several phases of catechol-oxidation current: the current increased (to severalfold of the control values) followed by a decrease or even total disappearance, with a gradual return to control values. Under conditions of depletion of extracellular calcium by EGTA (5 microliters of 0.5 mol.l-1 KCl + 0.25 mol.l-1 EGTA in 10 mmol.l-1 HEPES, pH 7.4) K(+)-induced depolarization confirmed the key role of calcium in the release of catecholamine transmitters as well as that in processes regulating the uptake and metabolism of these transmitters. The voltammetric techniques used in the present study may be a useful tool in extending of our knowledge about the cellular mechanisms of stimulus-response coupling in nerve cells.     

ATP dependence of anion uptake by isolated vacuoles: requirement for excess Mg2+

Vacuoles were isolated from barley mesophyll protoplasts. [14C]Malate or 36Cl- were taken up from the surrounding medium. Uptake was only slightly increased in the presence of equimolar levels of ATP and Mg2+ (as magnesium gluconate). In the presence of excess Mg2+ in the medium, ATP-stimulated uptake of malate and chloride increased several-fold. Stimulation by excess Mg2+ was not observed for ATP-stimulated amino acid uptake by isolated vacuoles. Stimulation of uptake by excess Mg2+ was observed at all malate concentrations upto 10 mmol.l-1. The content of Mg2+ needed for half-maximum stimulation was about 3.5 mmol.l-1 in the presence of 1 mmol.l-1 ATP. The increase in Mg2+ concentration had no effect on the tonoplast ATPase activity.     

The changes in conformation of (Na+ K+)-ATPase from rat brain membranes are accompanied by changes of protein segment movements in the nanosecond range

Differential polarized phase fluorometry of fluorescein-5-isothiocyanate (FITC) showed that the activation of (Na,K)-ATPase in crude plasma membranes from rat brain by 10 mmol.l-1 K+ and 100 mmol.l-1 Na+ significantly increased the rotational relaxational rate (R) of enzyme-bound FITC. This increase was blocked by both ouabain (0.1 mmol.l-1) and vanadate (0.1 mmol.l-1). In the absence of ATP, R was increased less after adding of 10 mmol.l-1 K+ to the membranes. The shifts in the nanosecond movements of the protein segments measured as R during the activation of (Na,K)-ATPase suggest that this type of movement might be of some functional importance.     

Combined action of S-carvone and mild heat treatment on Listeria monocytogenes Scott A

The combined action of the plant-derived volatile, S-carvone, and mild heat treatment on the food-borne pathogen, Listeria monocytogenes, was evaluated. The viability of exponential phase cultures grown at 8 degrees C could be reduced by 1.3 log units after exposure to S-carvone (5 mmol l-1) for 30 min at 45 degrees C, while individual treatment with S-carvone or exposure to 45 degrees C for 30 min did not result in a loss in viability. Other plant-derived volatiles, namely carvacrol, cinnamaldehyde, thymol and decanal, were also found to reduce the viability of L. monocytogenes in combination with the same mild heat treatment at concentrations of 1.75 mmol l-1, 2.5 mmol l-1, 1.5 mmol l-1 and 2 mmol l-1, respectively. These findings show that essential oil compounds can play an important role in minimally processed foods, and can be used in the concept of Hurdle Technology to reduce the intensity of heat treatment or other individual hurdles.     

Bioenergetic Investigation of Action of Lithium to <Emphasis Type="Italic">Tetrahymena thermophila</Emphasis> bF5 by Microcalorimetry

Microcalorimetry was employed to investigate the action of Li(I) to aquatic ecosystem from the point view of bioenergetics. Tetrahymena thermophila BF5 was chosen as the model organism. The power-time curves of T. thermophila BF5 growth metabolism in the absence and presence of Li(I) were obtained. The corresponding thermokinetic parameters were derived. The generation time was calculated as 592.3 min, which was consistent with the biomass values. Low concentration of Li(I) (1-20 mmol l-1) stimulated the growth of T. thermophila BF5, whereas the inhibition effect was observed in high concentration (30-100 mmol l-1). The value of IC50 was 52.8 mmol l-1. In the concentration range of 30-100 mmol l-1, the growth rate constants (k) and the maximum heat out power (P max) decrease with the concentration of Li(I), whereas the heat output (Q) increases slightly compared to the control. Other than the classic mechanism of inositol-phosphate cycle, the involvement of mitochondria mechanism was discussed and suggested.     

Evaluating in vitro and in vivo the interference of ascorbate and acetaminophen on glucose detection by a needle-type glucose sensor.

The aim of this work was to assess, in vitro and in vivo, the interference of ascorbate and acetaminophen on glucose measurements by a needle-type glucose sensor detecting hydrogen peroxide generated during the enzymatic oxidation of glucose, and to ascertain whether the protection against interference by the membranes used in the construction of the electrode is feasible. The oxidation of ascorbate and acetaminophen on a platinum electrode set at a 650 mV potential yielded a current representing 75 +/- 5% and 25 +/- 6% of that generated by the oxidation of an equimolar concentration of hydrogen peroxide, respectively. The bias introduced by the presence of 100 mumol l-1 ascorbate on the reading of 5 mmol l-1 glucose by the complete sensor (electrode + membranes) would be minimal (approximately 0.4 mmol l-1). By contrast, the bias introduced by 200 mumol l-1 of acetaminophen (a plasma concentration easily reached in clinical practice) was about 7 mmol l-1. The sensor was implanted subcutaneously in anaesthetized rats (n = 3). Using the current generated in the presence of a plasma acetaminophen concentration of about 200 mumol l-1 for glucose monitoring would lead to a major underestimation (approx. 6 mmol l-1) of subcutaneous glucose concentrations.     

Investigations on the photosynthetic sulfur bacterium Chlorobium phaeobacteroides causing seasonal blooms in Lake Kinneret

Between May and December, the annual stratification period in Lake Kinneret, sulfide is formed and accumulates in the hypolimnion. In July-August a large population (up to 10(6) cells/mL) of green, photosynthetic, sulfur bacteria develops at the boundary of the oxidative and reductive zones of the water column lasting for 3--8 weeks. These bacteria were isolated from the lake and identified as Chlorobium phaeobacteroides. Optimal growth conditions included 160 mg S=L-1 and light intensities of 5--30 micron Einstein (micron E) m-2s-1. Glucose and acetate augmented growth when added to the mineral medium. The lowest light intensity which still supported growth was 0.3 micron E m-2s-1 when acetate was present and 1.0 micron E m-2s-1 when no organic substrate was present. Under complete darkness, either with or without organic substrate, the bacteria die. Photosynthetic activity was higher when no organic compound was added to the medium. Uptake of acetate was light-dependent. In the lake the photosynthetic activity of the bacteria is low because of the limited light intensity (0.3 micron E m-2s-1) at the bloom layer. It is suggested that the appearance and the disappearance of the bloom are caused by the influence of the daily internal seiche.     

Overproduction of laccase by a monokaryotic strain of Pycnoporus cinnabarinus using ethanol as inducer

AIMS: Laccase production by the monokaryotic strain Pycnoporus cinnabarinus ss3 was studied using ethanol as inducer in the culture medium. METHODS AND RESULTS: The effect of ethanol was tested at 10, 20, 30, 35 and 45 g l-1 and compared with that of ferulic acid, known until now as the most efficient inducer for laccase expression by P. cinnabarinus ss3. In the presence of 35 g l-1 ethanol, laccase activity (266 600 U l-1) and productivity (19 000 U l-1 day-1) were nine and fivefold higher compared with ferulic acid-induced cultures, and 155- and 65-fold higher compared with non-induced cultures, respectively. In vivo, ethanol added to the culture medium of P. cinnabarinus ss3 favoured a continuous and high expression of laccase gene. Under these conditions, P. cinnabarinus ss3 produced preferentially the isoenzyme LAC I. Ethanol added in vitro to the purified P. cinnabarinus ss3 laccase typically inhibited the enzymatic activity. CONCLUSIONS: In spite of an initial inhibitory effect on mycelial growth, ethanol was shown to be a very strong inducer for laccase expression by P. cinnabarinus ss3 allowing an average yield of 1-1.5 g l-1 laccase. SIGNIFICANCE AND IMPACT OF THE STUDY: This study identified P. cinnabarinus ss3 as an outstanding producer of laccase in the presence of ethanol as inducer. Ethanol is an inexpensive agricultural by-product and the process is simple to scale-up for industrial production.     

Utilization of carbonate and ammonia-based treatments to eliminate Escherichia coli O157:H7 and Salmonella Typhimurium DT104 from cattle manure

AIMS: The objective of this study was to investigate alkaline treatments of cattle manure to kill coliforms, Escherichia coli O157:H7 and Salmonella Typhimurium DT104 based on their inhibition by carbonate ion and ammonia. METHODS AND RESULTS: Pure cultures of S. Typhimurium DT104 and E. coli O157:H7 strains were treated with sodium carbonate and ammonia to determine threshold inhibitory concentrations. Fresh cattle manure samples were inoculated with the same strains and their survival was determined after addition of sodium hydroxide, ammonium sulphate, sodium carbonate and/or urea. Control of CO and NH3 concentrations in manure by pH adjustment to 9.5 with sodium hydroxide to more than 5 and 30 mmol l-1, respectively, killed more than 106 cells g-1 in 7 days. Addition of sodium carbonate enhanced the killing effect of NaOH by increasing the CO and NH3 concentrations. Addition of 100 mmol l-1 urea, produced high levels of CO and NH3 and decreased all bacterial counts by at least 106 cells g-1 after 7 days. CONCLUSIONS: Reduction of food-borne pathogens in manure can be achieved by a combination of high concentrations of CO and NH3 which are pH-dependent parameters. SIGNIFICANCE AND IMPACT OF STUDY: Addition of urea could provide a simple manure treatment by combining both antimicrobial factors.     

Enhancement of biocontrol activity of yeasts by adding sodium bicarbonate or ammonium molybdate to control postharvest disease of jujube fruits

AIMS: To assess the potential of sodium bicarbonate and ammonium molybdate as additives in enhancing the biocontrol efficacy of Rhodotorula glutinis and Cryptococcus laurentii against blue mould in jujube fruits. METHODS AND RESULTS: Two yeasts at a concentration of 107 CFU ml-1, in combination with 238 mmol l-1 sodium bicarbonate or 15 mmol l-1 ammonium molybdate, showed a significant inhibition effect on blue mould of jujube fruits stored at 20 degrees C for 5 days. The colonizing ability of the yeasts in wounded sites was significantly decreased in the presence of ammonium molybdate. CONCLUSIONS: Combining R. glutinis or C. laurentii with sodium bicarbonate or ammonium molybdate provided a more effective control of postharvest disease than using the antagonistic yeasts or the chemicals alone. SIGNIFICANCE AND IMPACT OF THE STUDY: The addition of sodium bicarbonate or ammonium molybdate reduced the number of antagonists required to efficiently control disease of postharvest fruits, which could result in the reduction of costs.     

The Prospect of Purple Non-Sulfur (PNS) Photosynthetic Bacteria for Hydrogen Production: The Present State of the Art

Hydrogen is the fuel for the future, mainly due to its recyclability and nonpolluting nature. Biological hydrogen production processes are operated at ambient temperature and atmospheric pressures, thus are less energy intensive and more environmentally friendly as compared to thermochemical and electrochemical processes. Biohydrogen processes can be broadly classified as: photofermentation and dark fermentation. Two enzymes namely, nitrogenase and hydrogenase play an important role in biohydrogen production. Photofermentation by Purple Non-Sulfur bacteria (PNS) is a major field of research through which the overall yield for biological hydrogen production can be improved significantly by optimization of growth conditions and immobilization of active cells. The purpose of this paper is to review various processes of biohydrogen production using PNS bacteria along with several current developments. However, suitable process parameters such as carbon and nitrogen ratio, illumination intensity, bioreactor configuration and inoculum age may lead to higher yields of hydrogen generation using PNS bacteria.     

Inorganic compounds have dual effect on recombinant protein production: influence of anions and cations on serine alkaline protease production

AIMS: Investigation of concerted effects of cations, i.e. Mg2+ and Mn2+, in combination with their anions, i.e. sulphate, chloride and acetate (Ac), on the physiology of Bacillus licheniformis carrying pHV1431::subC to improve the fermentation medium for serine alkaline protease (SAP) production, whereupon, determination of the acid that can be used in pH control. METHODS AND RESULTS: The cell concentrations increased with the increase in MnSO4 and Mn(Ac)2 concentrations, and the highest values were obtained at Co(MnSO4) = 0.20 mmol l-1 and Co(Mn(CH3COO)2) = 4.0 mmol l-1, as 2.3 and 2.2 g l-1, respectively. However, Co(MnCl2) did not influence biomass concentration. SAP production was inhibited with MnCl2 after Co(MnCl2) = 0.60 mmol l-1, but with MnSO4 SAP production was inhibited drastically. Whereas, at high concentrations of Mn(Ac)2 SAP production increased and the highest activity was obtained as ASAP = 1285 U ml-1 at t = 65 h. With the Mg compounds, cell concentrations increased with the increase in the concentrations of MgSO4, MgCl2 and Mg(Ac)2; and the anions did not show any influence on the cell growth. Similar to the results of Mn compounds, the glucose consumption rate increased with the increase in MgSO4 and MgCl2 concentrations; contrariwise, decreased with the increase in Mg(Ac)2 concentrations, due to the use of acetate as the second carbon source. Co(MgSO4) = 0.40 mmol l-1, Co(MgCl2) = 1.60 mmol l-1 and Co(Mg(Ac)2) = 0.40 mmol l-1 were the optimum concentrations separately, and the highest SAP activity was obtained with Mg(Ac)2 as ASAP = 1338 U ml-1 at t = 47 h. Consequently, ion acetate and its acid HAc appear, respectively, as the superior anion for the essential cations and the control agent for pH control in the bioreactor. Finally, optimum initial concentrations and the concerted effects of Mg(Ac)2 and Mn(Ac)2 were investigated, and the optimum concentrations were found respectively as 0.40 and 0.80 mmol l-1, while the maximum activity was obtained as ASAP = 1010 U ml-1 at a shortened cultivation time of t = 39 h. CONCLUSIONS: Mn(Ac)2 and Mg(Ac)2 together enhanced the cell formation and SAP synthesis rates, moreover, SAP synthesis started at an earlier cultivation time. SIGNIFICANCE AND IMPACT OF THE STUDY: Each inorganic compound with its cation and anion has dual effect on the metabolism. Mg2+ and Mn2+ at their specific concentrations influence the regulation of the pathways that might cause better coupling of supply and demand for the amino acids on the basis of the amino acid composition of the enzyme molecule.     

Improvement of xylitol production by Candida guilliermondii FTI 20037 previously adapted to rice straw hemicellulosic hydrolysate

AIMS: To evaluate a simple and economical technique to improve xylitol production using concentrated xylose solutions prepared from rice straw hemicellulosic hydrolysate. METHODS AND RESULTS: Experiments were carried out with rice straw hemicellulosic hydrolysate containing 90 g l-1 xylose, with and without the addition of nutrients, using the yeast Candida guilliermondii previously grown on the hydrolysate (adapted cells) or on semi-defined medium (unadapted cells). By this method, the yield of xylitol increased from 17 g l-1 to 50 g l-1, and xylose consumption increased from 52% to 83%, after 120 h of fermentation. The xylitol production rates were very close to that (0.42 g l-1 h-1) attained in a medium simulating hydrolysate sugars. CONCLUSION: Yeast strain adaptation to the hydrolysate showed to be a suitable method to alleviate the inhibitory effects of the toxic compounds. Adapted cells of Candida guilliermondii can efficiently produce xylitol from hydrolysate with high xylose concentrations. SIGNIFICANCE AND IMPACT OF THE STUDY: Yeast adaptation helps the bioconversion process in hydrolysate made from lignocellulosic materials. This low-cost technique provides an alternative to the detoxification methods used for removal of inhibitory compounds. In addition, the use of adapted inocula makes it possible to schedule a series of batch cultures so that the whole plant can be operated almost continuously with a concomitant reduction in the overall operation time.