Ferric iron-reducing Shewanella putrefaciens and N2-fixing Bradyrhizobium japonicum with uptake hydrogenase are unable to oxidize atmospheric H2

Abstract Bradyrhizobium japonicum and Shewanella putrefaciens were unable to oxidize hydrogen at atmospheric concentrations (0.55 ppmv), neither in suspension nor when added to sterile soil. The K _m-value of S. putrefaciens for H₂ (39 ppmv in gas phase, 0.22 μM in aqueous phase), using Fe(III) as electron acceptor, showed a 4–5-fold higher affinity for H₂ than that of B. japonicum (1200 ppmv; 0.84 μM) or other hydrogen-oxidizing bacteria. However, the V _max (4.54 fmol H₂ h⁻¹ cell ⁻¹) and threshold (> 0.5 ppmv; 0.35 nM) of S. putrefaciens and the V _max (7.19 fmol H₂ h⁻¹ cell⁻¹) and threshold (> 0.5 ppmv; 0.35 nM) of B. japonicum were in the same order of magnitude as data for Knallgas bacteria from relevant literature. To enable hydrogen oxidation in soil the soil-samples with S. putrefaciens even had to be supplemented with Fe(III). Fresh soil, on the other hand, oxidized hydrogen very efficiently below atmospheric mixing ratios, demonstrating that there must be other oxidation activities in soil.     

从两类不同培养基上分离的土壤细菌的特征比较

通过对两类不同培养基上分离的土壤细菌的生理特征的比较,发现两组细菌的种群有所不同.芽孢杆菌主要生长在宫营养的培养基上,而很难在低营养的培养基上生长.假单孢杆菌和棒杆类型的细菌则在两类培养基上均可生长.这与两类细菌对环境的适应能力有关.     

氢氧化细菌分离、筛选及促生机制研究进展

氢氧化细菌是一类以氢作为电子供体,通过氧化H2获得能量并同化CO2的无机化能自养菌。近年来,发现作为豆科作物根际主要生理类群的氢氧化细菌促生作用明显,因而受到各国学者的广泛关注。氢氧化细菌在农业、环境保护和人类健康等领域有巨大的应用潜力。目前,由于人们对氢氧化细菌的分类、分离和筛选技术的认识不足,严重影响了氢氧化细菌的研究进程和水平。概述氢氧化细菌的分离及筛选方法,并着重论述了氢氧化细菌促生作用及促生机理的最新研究进展,最后探讨其应用前景。随着研究的深入,氢氧化细菌将受到各国学者的广泛关注。     

固定化光合细菌产氢过程的基质利用动力学

研究了固定化荚膜红假单胞菌386、红假单胞菌D两菌株产氢过程基质利用的动力学特性。基质利用与产氢过程以不同的速率进行．琼脂包埋固定化细胞的产氢能力高于海藻酸钙固定化细胞,但最大产氢活性的进程迟于后者。固定化D菌株利用葡萄糖的动力学遵循一级反应,其反应常数K值为1．2×10　2h-1。宏观动力学分析表明,利用基质的产氢过程属于反应控制,扩散传质过程不构成控速步骤。386和D两个菌株固定化细胞生物反应器连续产氢系统,在以乳酸作基质时．平均产氢量分别可达到0．659L／d和0．457L／d,容积(液相)产氢率接近1．OL／L·d。     

Characterization of populations of aerobic hydrogen-oxidizing soil bacteria

Abstract Freshly isolated soil bacteria were screened for different characteristics of the H₂ metabolism without prior selection for growth on H₂. The bacteria were isolated from different grain size fractions of a neutral meadow cambisol and an acidic forest cambisol, and then tested (1) for the ability to oxidize H₂, (2) for chemolithoautotrophic growth on H₂ as sole electron donor and energy source, (3) for DNA-DNA-hybridization with two hydrogenase gene fragments from Alcaligenes eutrophus and Rhizobium leguminosarum , and (4) for reduction of 2,3,5-triphenyl-2H-tetrazoliumchloride (TTC) in the presence of H₂. Many (65–90%) of the isolates were able to reduce TTC, but only 30–65% were actually able to oxidize H₂ indicating that the TTC test was not a specific characteristic for H₂ oxidation ability. The TTC test was only reliable in pure cultures of known bacteria with optimized test conditions, here shown for Alcaligenes eutrophus, Bradyrhizobium japonicum and Nocardia opaca , but not in mixed cultures of unknown bacteria. Still less (< 30%) of the isolates were able to grow chemolithoautotrophically indicating that culturable aerobic bacteria with the ability for H₂ oxidation are more abundant than bacteria with the ability for chemolithoautotrophic growth. The DNA-DNA-hybridization test failed to detect many of the bacteria with H₂ oxidation activity, probably since the hydrogenase genes present in the isolates were too diverse to be all detected by the DNA probes applied.     

Kinetic studies on elemental sulfur oxidation by Acidithiobacillus ferrooxidans: sulfur limitation and activity of free and adsorbed bacteria

The kinetics of sulfur oxidation by Acidithiobacillus ferrooxidans in shaking flasks and a 10-L reactor was studied. The observed linearity of growth and sulfur oxidation was explained by sulfur limitation. Total cell yield was not significantly different for exponential growth as compared to growth during the sulfur-limiting phase. Kinetic studies of sulfur oxidation by growing and nongrowing bacteria indicated that both free and adsorbed bacteria oxidize sulfur. Changes in the number of free bacteria rather than cells adsorbed on sulfur were better predictors of the kinetics of sulfur oxidation, indicating that the free bacteria were performing sulfur oxidation. The active growth phase always followed adsorption of bacteria on sulfur; however, the special metabolic role of adsorbed bacteria was unclear. Their activity in sulfur solubilization was considered.     

Methionine regulates copper/hydrogen peroxide oxidation products of Abeta.

Metal-catalysed oxidation (MCO) may play a causative role in the pathogenesis of Alzheimer's disease (AD). Amyloid beta peptide (Abeta), the major biomarker of AD, in the presence of copper ions reduces Cu(2+) to Cu(+) and catalyses the formation of H(2)O(2) that subsequently induces radicals through Fenton chemistry. Abeta is also subject to attack by free radicals, where the presence of Cu(2+) in conjunction with H(2)O(2) catalyses oxygenation, primarily at the methionine sulfur atom. This work investigates MCO of Abeta, to gain further insight into the role of oxidative stress in AD. By combining a fluorescence assay with gel electrophoresis to monitor MCO reactions of Abeta (1-28) in the presence and absence of methionine it was determined that methionine can both protect some residues against MCO and promote the oxidation of Tyr(10) specifically. Electrospray ionization mass spectrometric analysis of methionine MCO products indicated the formation of methionine sulfoxide, methionine sulfone and related hydroxylated products. Similar products could be formed from the oxidation of Met(35) of Abeta and may relate to changes in properties of the peptide following MCO.     

植物次生物质对烟青虫和棉铃虫食物利用及中肠解毒酶活性的影响

用分别添加0-5%(干重比)棉酚、烟碱、番茄苷和辣椒素4种植物次生物质的人工饲料饲养烟青虫Helicoverpa assulta和棉铃虫H. armigera 5龄幼虫48 h,测定这些次生物质对烟青虫和棉铃虫的营养效应和中肠谷胱甘肽S-转移酶（GST）及羧酸酯酶（CarE）活性的影响。结果表明：在实验浓度下,棉酚可显著降低烟青虫的相对消化率,但对棉铃虫却有助食作用;番茄苷抑制烟青虫的取食和生长,对其近似消化率和食物利用率也有显著的抑制作用,但食物转化率有明显升高,对棉铃虫的各营养指标无显著影响;烟碱对烟青虫和棉铃虫的相对生长率均无影响;辣椒素使烟青虫的取食量有大幅度的提高,对棉铃虫的取食量无影响,但引起其相对消化率的提高。由此可见,棉铃虫对4种次生物质有普遍的适应性,而烟青虫只对寄主植物所含的烟碱和辣椒素有较好的适应性。烟青虫和棉铃虫幼虫中肠CarE活性不受4种次生物质的影响,烟碱和辣椒素对烟青虫GST有显著的诱导作用,番茄苷对烟青虫GST活性则有抑制作用,4种次生物质对棉铃虫GST均无显著影响。     

Kinetics of iron oxidation by Leptospirillum ferriphilum dominated culture at pH below one

The kinetics of ferrous iron oxidation by Leptospirillum ferriphilum (L. ferriphilum) dominated culture was studied in the concentration range of 0.1-20 g Fe(2+)/L and the effect of ferric iron (0-60 g Fe(3+)/L) on Fe(2+) oxidation was investigated at pH below one. Denaturing gradient gel electrophoresis of PCR amplified 16S rRNA genes followed by partial sequencing confirmed that the bacterial community was dominated by L. ferriphilum. In batch assays, Fe(2+) oxidation started without lag phase and the oxidation was completed within 1 to 60 h depending on the initial Fe(2+) concentration. The specific Fe(2+) oxidation rates increased up to around 4 g/L and started to decrease at above 4 g/L. This implies substrate inhibition of Fe(2+) oxidation at higher concentrations. Haldane equation fitted the experimental data reasonably well (R(2) = 0.90). The maximum specific oxidation rate (q(m)) was 2.4 mg/mg VS . h, and the values of the half saturation (K(s)) and self inhibition constants (K(i)) were 413 and 8,650 mg/L, respectively. Fe(2+) oxidation was competitively inhibited by Fe(3+) and the competitive inhibition constant (K(ii)) was 830 mg/L. The time required to reach threshold Fe(2+) concentration was around 1 day and 2.3 days with initial Fe(3+) concentration of 5 and 60 g/L, respectively. The threshold Fe(2+) concentration, below which no further Fe(2+) oxidation occurred, linearly increased with increasing initial Fe(2+) and Fe(3+) concentrations. Fe(2+) oxidation proceeds by L. ferriphilum dominated culture at pH below 1 even in the presence of 60 g Fe(3+)/L. This indicates potential of using and biologically regenerating concentrated Fe(3+) sulfate solutions required, for example, in indirect tank leaching of ore concentrates.     

Copper (II) complex-catalyzed oxidation of NADH by hydrogen peroxide

Among various metal ions of physiological interest, Cu²⁺ is uniquely capable of catalyzing the oxidation of NADH by H₂O₂. This oxidation is stimulated about fivefold in the presence of imidazole. A similar activating effect is found for some imidazole derivatives (1-methyl imidazole, 2-methyl imidazole, andN-acetyl-L-histidine). Some other imidazole-containing compounds (L-histidine,L-histidine methyl ester, andL-carnosine), however, inhibit the Cu²⁺-catalyzed peroxidation of NADH. Other chelating agents such as EDTA andL-alanine are also inhibitory. Stoichiometry for NADH oxidation per mole of H₂O₂ utilized is 1, which excludes the possibility of a two-step oxidation mechanism with a nucleotide free-radical intermediate. About 92% of the NADH oxidation product can be identified as enzymatically active NAD⁺. D₂O, 2,5-dimethylfuran, and 1,4-diazabicyclo [2.2.2]-octane have no significant effect on the oxidation, thus excluding¹O₂ as a mediator. Similarly, OH· is also not a likely intermediate, since the system is not affected by various scavengers of this radical. The results suggest that a copper-hydrogen peroxide intermediate, when complexed with suitable ligands, can generate still another oxygen species much more reactive than its parent compound, H₂O₂.     

Biological sulfide oxidation using autotrophic Thiobacillus sp.: evaluation of different immobilization methods and bioreactors

Aims:  Evaluation of various immobilization methods and bioreactors for sulfide oxidation using Thiobacillus sp. was studied.
Methods and Results:  Ca-alginate, K-carrageenan and agar gel matrices (entrapment) and polyurethane foam and granular activated carbon (adsorption) efficacy was tested for the sulfide oxidation and biomass leakage using immobilized Thiobacillus sp. Maximum sulfide oxidation of 96% was achieved with alginate matrix followed by K-carrageenan (88%). Different parameters viz. alginate concentration (1%, 2%, 3%, 4% and 5%), CaCl₂ concentration (1%, 2%, 3%, 4% and 5%), bead diameter (1, 2, 3, 4 and 5 mm), and curing time (1, 3, 6, 12 and 18 h) were studied for optimal immobilization conditions. Repeated batch experiments were carried out to test reusability of Ca-alginate immobilized beads for sulfide oxidation in stirred tank reactor and fluidized bed reactor (FBR) at different sulfide concentrations.
Conclusions:  The results proved to be promising for sulfide oxidation using Ca-alginate gel matrix immobilized Thiobacillus sp. for better sulfide oxidation with less biomass leakage.
Significance and Impact of the Study:  Biological sulfide oxidation is gaining more importance because of its simple operation. Present investigations will help in successful design and operation of pilot and industrial level FBR for sulfide oxidation.     

Soil microbial activities and litter decomposition related to altitude

Summary On a southern slope in the Austrian Central Alps (Hohe Tauern Mountains) at altitudes of 2550 m, 1920 m and 1650 m above sea level, respectively, microbial activities were investigated by measuring the decomposition of litter, the cellulase and xylanase activities, CO₂-evolution and the cell counts of viable non differentiated and cellulolytic bacteria. After one year 46% of litter exposed was decomposed at an altitude of 2550 m, 76% at 1920 m and 86% at 1560 m. Investigations with litter bags of different mesh sizes (25 μm and 1000 μm) revealed that small soil animals (<1 mm) did not significantly influence the decomposition of litter at different altitudes and in different vegetation types. The enzymatic activities and the CO₂-evolution of soils decreased with increasing altitude. Plate counts of bacteria from soils at the alpine zone (2550 m) and the tall grass meadow (1650 m) indicated that in some cases the lower metabolic activities caused by bad climatic conditions were compensated by an increase in cell numbers. This work was supported by the Austrian MaB-Hochgebirgsprogramm, Alpine ?kosysteme     

Microbial manganese(II) oxidation in the marine environment: a quantitative study

A great number of important chemical reactions that occur in the environment are microbially mediated. In order to understand the kinetics of these reactions it is necessary to develop methods to directly measure in situ reaction rates and to develop models to help elucidate the mechanisms of microbial catalysis. The oxidation of Mn(II) in a zone above the O₂/H₂S interface in Saanich Inlet, B.C., Canada is one such reaction. We present here a method by which in situ rates of microbial Mn(II) oxidation are measured and a model based on our experimental results to describe the general mechanism of Mn(H) oxidation. We propose a two step process in which Mn(II) is first bound by a site on the bacterial surface and then oxidized. The model is analogous to the Langmuir isotherm model for surface catalyzed gas reactions or the Michaelis-Menten model for enzyme kinetics. In situ Mn(II) oxidation rates were measured during five cruises to Saanich Inlet during the summers of 1983 and 1984. We use the model to calculate the apparent equilibrium binding constant (K_s 0.18 M), the apparent half saturation constant for biological Mn(H) oxidation (K_m = 0.22 to 0.89 M), the maximum rate of Mn(II) oxidation (V_max = 3.5 to 12.1 nM·h^-1) and the total microbial surface binding site concentration ( E 51 nM). V_max for Mn(II) oxidation agrees with the rates calculated from the value of the flux of Mn(II) to the oxidizing zone using the Mn(II) gradient and estimates of the eddy diffusion coefficient. This consistancy verifies our methodology and indicates that the rate of Mn(II) oxidation is nearly equal to the (V_max for the reaction. We conclude that in this environment the Mn(II) oxidation rate is more a function of the total number of surface binding sites than the Mn(H) concentration.Contribution #1601 from the School of Oceanography, Univ. of Washingtoncorresponding author     

The oxidative products of methionine as site and content biomarkers for peptide oxidation

Biomarkers for peptide/protein oxidation under oxidative stress (OS) hold both incredible application potential as well as significant challenges. In this article, liquid chromatography and mass spectrometry were applied to establish a new method for evaluating the oxidation site and degree of peptide oxidized, with its oxidative product serving as biomarker. In the three model peptides, peptide FMRF (containing a methionine) was prone to undergo oxygen addition under UV/H₂O₂ oxidization, forming a sulfoxide (FM(O)RF) with a stable chromatographic peak separate from the model peptides. The oxidation content of FMRF, expressed as S_FM(O)RF/(S_FM(O)RF + S_FMRF), is positively correlated with oxidation time. Based on sequence analysis of FM(O)RF, the oxidation mechanism (site and extent) of FMRF under UV/H₂O₂ oxidization was explicitly clarified. By comparing the specific injury to each model peptide, we found that the oxidative products of Met‐containing peptides are good biomarkers for OS. This research not only expands the range of biomarkers for OS, but also provides an efficient and accurate method for evaluating oxidation damage to peptides and even proteins. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd.     

Determination of sulphite in mineral soils

Summary Two methods were evaluated to determine the concentration of sulphite in sandy loam soils. Use of p-rosaniline-formaldehyde reagent and sodium tetrachloromercurate as extractant proved effective only if a wide soil to extractant ratio was employed. Use of Ellman's reagent in conjunction with this extractant was however, unsatisfactory. Soils exposed to heavy atmospheric pollution differed in sulphite concentration only marginally compared with relatively unpolluted soils. The effect of soil water content on the stability of sulphite in soil was also determined.     

Capacity of Irpex lacteus and Pleurotus ostreatus for decolorization of chemically different dyes

The rate and efficiency of decolorization of poly R-478- or Remazol Brilliant Blue R (RBBR)-containing agar plates (200 μg g⁻¹) were tested to evaluate the dye degradation activity in a total of 103 wood-rotting fungal strains. Best strains were able to completely decolorize plates within 10 days at 28 °C. Irpex lacteus and Pleurotus ostreatus were selected and used for degradation of six different groups of dyes (azo, diazo, anthraquinone-based, heterocyclic, triphenylmethane, phthalocyanine) on agar plates. Both fungi efficiently degraded dyes from all groups. Removal of RBBR, Bromophenol blue, Cu-phthalocyanine, Methyl red and Congo red was studied with I. lacteus also in liquid medium. Within 14 days, the following color reductions were attained: RBBR 93%, Bromophenol blue 100%, Cu-phthalocyanine 98%, Methyl red 56%, Congo red 58%. The ability of I. lacteus to degrade RBBR spiked into sterile soil was checked, the removal being 77% of the dye added within 6 weeks. The capacity of selected white rot fungal species to remove efficiently diverse synthetic dyes from water and soil environments is documented.     

Requirement of intracellular free thiols for hydrogen peroxide-induced hypertrophy in cardiomyocytes

Reactive oxygen species (ROS) are by-products of aerobic metabolism and are implicated in the pathogenesis of several diseases. H(2)O(2) produces oxidative stress and acts as a second messenger in several cell types. We tested whether the effect of H(2)O(2) on cellular events could be altered by changes in the intracellular redox status in a cardiomyocyte cell line. Using flow cytometric measurements, we found that adding H(2)O(2) induced hypertrophy in control cells in a time-dependent manner. Pre-incubation of the cells with buthionine sulfoximine (BSO), an inhibitor of de novo GSH synthesis, induced increase in the number of cells of small sizes by the addition of H(2)O(2) as compared to non-BSO pre-incubated control cells, and exacerbated the decrease in viability. Total thiol and GSH levels in H9c2 cells pre-incubated with BSO were about 75 and 30% of control, respectively, and GSH levels fell to below the limitation of detection after the addition of H(2)O(2), although total thiol levels were not markedly decreased. In the cells pre-incubated with BSO, hypertrophy was not observed by the addition of H(2)O(2) at any level of concentration. N-acetyl-L-cysteine and cysteine not only prevented increase in the number of cells of small sizes caused by H(2)O(2) but also induced hypertrophy in cells pre-incubated with BSO. These results suggest that the intracellular free thiol levels determine whether cell death or hypertrophy occurs in cardiomyocytes in the presence of H(2)O(2). On the other hand, the hypertrophied cells did not become larger by adding H(2)O(2), but had high levels of cellular GSH, suggesting the possibility that the hypertrophied cells have tolerance to oxidative stress.     

Growth kinetics of the photosynthetic bacterium Chlorobium thiosulfatophilum in a fed-batch reactor

Hydrogen sulfide dissolved in water can be converted to elementary sulfur or sulfate by the photosynthetic bacterium Chlorobium thiosulfatophilum. Substrate inhibition occurred at sulfide concentrations above 5.7 mM. Light inhibition was found at average light intensities of 40,000 lux in a sulfide concentration of 5 mM, where no substrate inhibition occurred. Light intensity, the most important growth parameter, was attenuated through both scattering by sulfur particles and absorption by the cells. Average cell and sulfur particle sizes were 1.1 and 9.4 mum, respectively. Cells contributed 10 times as much to the turbidity as sulfur particles of the same weight concentration. The light attenuation factor was mathematically modeled, considering both the absorption and scattering effects based on the Beer-Lambert law and the Rayleigh theory, which were introduced to the cell growth model. Optimal operational conditions relating feed rate vs. light intensity were obtained to suppress the accumulation of sulfate and sulfide and save light energy for 2- and 4-L fed-batch reactors. Light intensity should be greater for the same performance (H(2)S removal rate/unit cell concentration) in larger reactors due to the scaleup effect on light transmission. Knowledge of appropriate growth kinetics in photosynthetic fed-batch reactors was essential to increase feed rate and light intensity and therefore cell growth. A mathematical model was developed that describes the cell growth by considering the light attenuation factor due to scattering and absorption and the crowding effect of the cells. This model was in good agreement with the experimental results. (c) 1992 John Wiley & Sons, Inc.