Noncompetitive inhibition by L-cysteine and activation by L-glutamate of the iron-oxidizing activity of a mixotrophic iron-oxidizing bacterium strain OKM-9

A mesophilic, mixotrophic iron-oxidizing bacterium strain OKM-9 uses ferrous iron as a sole source of energy and L-glutamate as a sole source of cellular carbon. Uptake of L-glutamate into OKM-9 cells is absolutely dependent on ferrous iron oxidation. Thus, the Fe(2+)-dependent L-glutamate uptake system of strain OKM-9 is crucial for the bacterium to grow mixotrophically in iron medium with L-glutamate. The relationship between iron oxidation and L-glutamate transport activities was studied. Iron oxidase containing cytochrome a was purified 9-fold from the plasma membrane of OKM-9. A purified iron oxidase showed one rust-colored band following disc gel electrophoresis after incubation with Fe(2+). The Fe(2+)-dependent L-glutamate transport system was also purified 14.5-fold from the plasma membrane using the same purification steps as for iron oxidase. Fe(2+)-dependent L-glutamate and L-cysteine uptake activities of OKM-9 were 0.36 and 0.24 nmol/mg/min, respectively, when a concentration of 18 mM of these amino acids was used as a substrate. Both uptake activities were completely inhibited by potassium cyanide (KCN), suggesting that cytochrome a in the iron oxidase is involved in the transport process. The iron-oxidizing activity of strain OKM-9 was activated 1.7-fold by 80 mM L-glutamate. In contrast, the activity was noncompetitively inhibited by L-cysteine. The Michaelis constant of iron oxidase for Fe(2+) was 12.6 mM and the inhibition constant for L-cysteine was 41.6 mM. A marked inhibition of iron oxidase by 50 mM L-cysteine was completely reversed by the addition of 60 mM L-glutamate. The results suggest the possibility that iron oxidase has a binding site for L-cysteine and the cysteine first bound to the iron oxidase was replaced by the added L-glutamate.     

Mass balance and kinetic analysis of anaerobic microbial dechlorination of pentachlorophenol in a continuous flow column

A mass balance and kinetic investigation of anaerobic dechlorination of pentachlorophenol (PCP) was undertaken using an enriched microbial consortium in a laboratory scale continuous flow column, as a model microbial permeable reactive barrier. The chlorine balance showed that 50 µM PCP was largely dechlorinated to phenol with the formation of a small quantity of 3-chlorophenol as an intermediate metabolite (hydraulic retention time 7.6 days), and the chlorine removal efficiency reached 36 µM d^-1. When the initial PCP concentration was increased to 100 µM the chlorine removal efficiency increased 1.5 times. However, the dechlorination activity disappeared after 7.4 pore volumes (58 days), demonstrating the susceptibility of the dechlorination culture to high concentrations of PCP. Lactate released hydrogen as an electron donor during PCP dechlorination, with acetate, propionate, CO₂ and CH₄ as byproducts. The carbon balance showed that some of the organic carbon source (PCP, lactate) in the influent was converted to gas and utilized for biomass growth in addition to organic metabolites. The kinetic study was conducted in a batch culture and yielded 1.99 mg l^-1 biomass growth per unit of chlorine consumption (µM). The Monod equation was well fitted to the specific growth rate of 1.38 d^-1 and a half saturation constant of 0.29 µM. The organic chlorine removal rate in the batch culture was consistent with the results in the flow column, indicating the feasibility of and potential for in situ estimation and prediction through batch culture studies.     

Multiresidue determination of quinolones in animal and fishery products by HPLC

A simple and rapid multiresidue method was developed for the determination of twelve quinolones (ciprofloxacin, danofloxacin, difloxacin, enrofloxacin, flumequine, marbofloxacin, nalidixic acid, norfloxacin, ofloxacin, orbifloxacin, oxolinic acid and sarafloxacin) in muscle, liver, chicken eggs, milk, prawn and rainbow trout.The quinolones were extracted from a sample with acetonitrile-water (95 : 5). A fifth part of the filtered extract was diluted with water to keep the acetonitrile ratio at ca. 60%, and passed through a C18 mini-column. The eluate was evaporated to dryness, and the residues were dissolved in methanol-water (30 : 70) for HPLC analysis.The quinolones were separated on a Inertsil ODS-3V column (4.6 mm i.d.x250 mm) with a gradient system of 0.1% phosphoric acid-acetonitrile as the mobile phase, with fluorescence detection.No interfering peak was found on the chromatograms of animal and fishery products, except for milk. The recoveries of the quinolones were over 60% from the animal and fishery products fortified at 0.1 microg/g, and the quantification limits of the quinolones were 0.005 microg/g. This proposed method was found to be effective and suitable for the screening of the quinolones in animal and fishery products.     

Dimethylamine-modified waste paper for the recovery of precious metals

Waste newsprint paper was modified with dimethylamine (DMA) to obtain a tertiary amine type adsorption gel called DMA-paper gel. This new derivative was investigated for adsorption, from hydrochloric acid medium, of gold, palladium, and platinum as well as some base metals. The gel exhibited selectivity only for precious metals with a remarkably high capacity for Au(III), i.e., 4.6 mol/kg dry gel and comparable capacities, i.e., 2.1 and 0.9 mol/kg for Pd(II) and Pt(IV), respectively. Also, Au(III) was reduced to the elemental form during adsorption. Furthermore, column adsorption and subsequent elution of the adsorbed metal ions by acidic thiourea revealed encouraging recoveries (approximately 90%), thus enhancing the scope of the gel for effective preconcentration, separation, and recovery of precious metals. The effectiveness of recovery of precious metals from real industrial liquor was also tested, and it showed highly encouraging results with respect to the stability of the gel in the harsh medium, and selectivity for the targeted metal ions in the presence of excess of other metal ions.     

Polysulfide reduction using sulfate-reducing bacteria in a photocatalytic hydrogen generation system

A hydrogen generation process using photocatalytic reactions has been proposed. In this process, hydrogen sulfide is a source of hydrogen and is turned into polysulfide. In order to establish the cyclic operation of a photocatalytic hydrogen generation system, it is necessary to convert polysulfide back into hydrogen sulfide with a small energy input. This paper proposes the use of sulfate-reducing bacteria (SRB) for the regeneration of hydrogen sulfide. Batch cultivation of natural source SRB samples were carried out using a culture medium containing polysulfide as the only sulfur compound source. SRB produced hydrogen sulfide from several kinds of polysulfide sources, including a photocatalytic hydrogen generation-produces solution. Production lag phase and production rate of hydrogen sulfide were affected by initial polysulfide concentration. SRB activity was inhibited at high initial polysulfide concentrations. SRB enrichment culture T2, exhibited the highest hydrogen sulfide production rate, and was able to utilize several kinds of organic matter as the electron donor. The results suggest the possibility of using large biomass sources, such as sewage sludge and the raw garbage in a hydrogen generation system. We developed speculative estimates that an SRB based hydrogen generation system is feasible.     

Inactivation of Bacillus spores by the combination of moderate heat and low hydrostatic pressure in ketchup and potage

The combination effect of moderate heat and low hydrostatic pressure (MHP) on the reduction of Bacillus subtilis, Bacillus coagulans and Geobacillus stearothermophilus spores in food materials (potage and ketchup) was investigated. These bacterial spores were suspended in potage (pH 7), acidified potage (pH 4), neutralized ketchup (pH 7) and ketchup (pH 4). The suspensions were treated with and without pressure (100 MPa) and temperatures of 65-85 degrees C for 3 to 12 h. The bacterial spores were inactivated by 4-8 log cycles during MHP treatment in potage, acidified potage and ketchup, whereas the spores were highly resistant to long time heat treatment in potage and neutralized ketchup. The degrees of spore destruction were mostly dependent on pH and medium composition during MHP treatment. The inactivation effect in MHP treatment was higher at the pH 7 than at pH 4 both in ketchup and potage. The bacterial spores showed higher inactivation in potage than ketchup during MHP treatment.     

Screening method for the determination of 199 pesticides in agricultural products by gas chromatography/ion trap mass spectrometry (GC/MS/MS)

A screening method is described for determining 200 pesticides, except dimethipin, divided into four groups by means of gas chromatography/tandem mass spectrometry (GC/MS/MS) using an ion trap mass spectrometer equipped with automated gain control (AGC). The quantitation limit for 194 pesticides was 0.01 mg/kg on a crop basis, except for allidochlor, dimethoate, hexythiazox, methamidophos and triadimenol. The calibration curve of each pesticide was linear in the range of 0.04-5.0 microg/mL. One hundred and ninety-nine pesticides were added to matrix of potato, spinach, cabbage, apple, orange, soybean and unpolished rice at twice the limits of quantitation. The recoveries of 194 pesticides from all crops were satisfactory (50-150%) for screening purposes. Although some pesticides in apple and orange were not determined by selected ion monitoring (SIM) analysis at the limits of quantitation, all of them were identified by ion-trap GC/MS/MS at the same concentration. Thus, the ion trap GC/MS/MS technique is useful for the screening of residual pesticides present at low levels in agricultural products.     

Water‐saving approaches for improving wheat production

The greatest fear of global climate change is drought. World‐wide, 61% of countries receive rainfall of less than 500 mm annually; domestication of wheat first occurred in such a semiarid region of southwestern Asia, and it seems that wheat foods originally came from dryland gardens. Wheat plants respond to drought through morphological, physiological and metabolic modifications in all plant parts. At the cellular level, plant responses to water deficit may result from cell damage, whereas other responses may correspond to adaptive processes. Although a large number of drought‐induced genes have been identified in a wide range of wheat varieties, a molecular basis for wheat plant tolerance to water stress remains far from being completely understood. The rapid translocation of abscissic acid (ABA) in shoots via xylem flux, and the increase of ABA concentration in wheat plant parts correlate with the major physiological changes that occur during plant response to drought. It is widely accepted that ABA mediates general adaptive responses to drought. For a relatively determinate target stress environment, and with stable genotype × environment interaction, the probability for achieving progress is high. This approach will be possible only after we learn more about the physiology and genetics of wheat plant responses to water stress and their interactions. The difficulties encountered by molecular biologists in attempting to improve crop drought tolerance are due to our ignorance in agronomy and crop physiology and not to lack of knowledge or technical expertise in molecular biology. Copyright © 2005 Society of Chemical Industry     

Inhibitory effect of high concentrations of ferric ions on the activity of Acidithiobacillus ferrooxidans

The influence of high concentrations of ferric ions on the biochemical activity of Acidithiobacillus ferrooxidans was studied using intact cells. The specific oxidation rate of ferrous ions decreased with increasing ferric ion concentration. Lineweaver-Burk plots revealed typical competitive inhibition kinetics, because the slopes varied with the ferric ion concentration. A linear relationship between the slope and the square of the ferric ion concentration revealed that the iron-oxidizing enzyme system of A. ferrooxidans was competitively inhibited by about two molecules of ferric ion. The kinetic equation based on this inhibition model agreed with the experimental observation at a high ferric ion concentration where the bacterium is usually exposed in bioleaching and biooxidation plants.