Tuning of the FMN binding and oxido-reduction properties by neighboring side chains in Anabaena flavodoxin

Contribution of three regions (phosphate-binding, 50’s and 90’s loops) of Anabaena apoflavodoxin to FMN binding and reduction potential was studied. Thr12 and Glu16 did not influence FMN redox properties, but Thr12 played a role in FMN binding. Replacement of Trp57 with Glu, Lys or Arg moderately shifted E_ox/sq and E_sq/hq and altered the energetic of the FMN redox states binding profile. Our data indicate that the side chain of position 57 does not modulate E_ox/sq by aromatic stacking or solvent exclusion, but rather by influencing the relative strength of the H-bond between the N(5) of the flavin and the Asn58-Ile59 bond. A correlation was observed between the isoalloxazine increase in solvent accessibility and less negative E_sq/hq. Moreover, E_sq/hq became less negative as positively charged residues were added near to the isoalloxazine. Ile59 and Ile92 were simultaneously mutated to Ala or Glu. These mutations impaired FMN binding, while shifting E_sq/hq to less negative values and E_ox/sq to more negative. These effects are discussed on the bases of the X-ray structures of some of the Fld mutants, suggesting that in Anabaena Fld the structural control of both electron transfer steps is much more subtle than in other Flds.     

Drought’s impact on Ca,Fe, Mg,Mo and S concentration and accumulation patterns in the plants and soil of a Mediterranean evergreen <Emphasis Type="Italic">Quercus </Emphasis><Emphasis Type="Italic">ilex</Emphasis> forest

We conducted a 6-year field manipulation drought experiment in an evergreen Quercus ilex forest where we simulated the drought predicted by GCM and ecophysiological models for the coming decades (an average of 15% soil moisture reduction). We thereby tested the hypothesis that enhanced drought will change Ca, Fe, Mg, Mo and S availability, concentrations and accumulation patterns in Mediterranean ecosystems. The strongest effects of drought occurred in the soil. Drought increased the total soil concentrations of S, the soil extract concentrations of Fe, Mg and S, the Mg saturation in the soil exchangeable complex and tended to increase the percentage base saturation of the soil exchangeable complex. These increased soil concentrations were related to a decrease of plant uptake capacity and not to an increase of soil enzyme activity, which in fact decreased under drier conditions. Drought increased leaf Mg concentrations in the three dominant species although only significantly in Quercus ilex and Arbutus unedo (20 and 14%, respectively). In contrast, drought tended to decrease Ca in Phillyrea latifolia (18%) and Ca and Fe concentrations in the wood of all three species. Drought increased Ca and Fe concentrations in the roots of Quercus ilex (26 and 127%). There was a slight general trend to decrease total biomass accumulation of nutrients that depend on water flux such as Mg, Fe and S. This effect was related to a decrease of soil moisture that reduced soil flow, and to a decrease in photosynthetic capacity, sap flow, transpiration and growth, and therefore plant uptake capacity under drought observed in Quercus ilex and Arbutus unedo. On the contrary, drought increased Mo accumulation in aboveground biomass in Phillyrea latifolia and reduced Mo accumulation in Arbutus unedo by reducing growth and wood Mo concentrations (51%). Phillyrea latifolia showed a great capacity to adapt to drier conditions, with no decrease in growth, an increase of Mo uptake capacity and a decrease in leaf Ca concentration, which was related to a decrease in transpiration under drought. The results indicate asymmetrical changes in species capacity to accumulate these elements, which are likely to produce changes in inter-specific competitive relations among dominant plant species and in their nutritional quality as food sources. The results also indicate that drought tended to decrease nutrient content in aboveground biomass, mainly through the decrease in growth and transpiration of the most sensitive species and caused an increase in the availability of these nutrients in soil. Thus, drought decreased the ecosystem’s capacity to retain Mg, Fe and S, facilitating their loss in torrential rainfalls.     

Effectiveness of sulfur with Acidithiobacillus and gypsum in chemical attributes of a Brazilian sodic soil

Gypsum and sulfur have been used as amendments for application in sodic and saline sodic soils, although gypsum is not effective in soil pH reduction. In this study the combined effects of elemental sulfur inoculated with Acidithiobacillus (S*) and gypsum (G) in chemical attributes of a Brazilian solodic soil was evaluated. The treatments consisted in addition of S* and G in various levels (0, 0.8, 1.6, 2.4, and 3.2 t ha⁻¹) and different mixing proportions (100:0, 75:25, 50:50, 25:75, and 100:0), acting during 15, 30, and 45 days. Sulfur inoculated with Acidithiobacillus (S*) markedly reduced soil pH in the leaching solution, especially when applied in the highest levels. Gypsum or sulfur applied individually was not satisfactory for soil reclamation. At 15 days of incubation Na⁺, Ca²⁺, and Mg²⁺ showed higher values in the leaching solution, and a marked decrease was observed in the leaching solution at 30 days. Reduction in soil electrical conductivity and in exchangeable Na⁺, Ca²⁺, and Mg²⁺ was observed and in a general way best results were achieved with S* : G in the ratio 50:50, using 2.4 and 3.2 t ha⁻¹. Sulfur with Acidithiobacillus was more effective than gypsum in decreasing soil pH, and sulfur applied with gypsum in the proportion 50:50 showed the best results in relation to exchangeable sodium and electrical conductivity and showed values below those used for classification as sodic soils.     

Bioleaching of heavy metals from contaminated soil using <Emphasis Type="Italic">Acidithiobacillus thiooxidans</Emphasis>: effect of sulfur/soil ratio

Bioleaching of heavy metals from contaminated soil was carried out using indigenous sulfur oxidizing bacterium Acidithiobacillus thiooxidans. Experiments were carried out by varying sulfur/soil ratio from 0.03 to 0.33 to evaluate the optimum ratio for efficient bioleaching of heavy metals from soil. The influence of sulfur/soil ratio on the bioleaching efficiency was assessed based on decrease in pH, increase in oxidation–reduction potential, sulfate production and solubilization of heavy metals from the soil. Decrease in pH, increase in oxidation–reduction potential and sulfate production was found to be better with the increase in sulfur/soil ratio. While the final pH of the system with different sulfur/soil ratio was in the range of 4.1–0.7, oxidation reduction potential varied from 230 to 629 mV; sulfate production was in the range of 2,786–8,872 mg/l. Solubilization of chromium, zinc, copper, lead and cadmium from the contaminated soil was in the range of 11–99%. Findings of the study will help to optimize the ratio of sulfur/soil to achieve effective bioleaching of heavy metals from contaminated soils.     

The role of sulfur assimilation and sulfur-containing compounds in trace element homeostasis in plants

Sulfur assimilation and production of sulfur-containing compounds are essential biological activities that play critical roles in many biological processes, including the role of sulfur containing compounds such as glutathione and phytochelatin in trace element homeostasis in plants. This review will discuss the role of sulfur assimilation and the biosynthesis of sulfur containing compounds in both mechanisms of trace element hyperaccumulation and heavy metal stress responses in plants.     

Newly Detected Specific Hydrogenation of the Conjugated Double Bond of Unsaturated Alkaloid Lactones by Aspergillus sp.

A new isolate of Aspergillus sp. hydrogenated the γ,δ-double bond of securinine (143 mg l⁻¹) to give 14,15-dihydrosecurinine at over 98% (w/w) yield after 8 h. It also hydrogenated the C11(13) double bond of 3-hydroxy-1(10),3,11(13)-guaiatriene-12,6-olide-2-one (HGT) (200 mg l⁻¹) to give 3-hydroxy-1(10),3-guaiadiene-12,6-olide-2-one with over 98% (w/w) conversion after 24 h.     

Deciphering the role of histidine 252 in mycobacterial adenosine 5'-phosphosulfate (APS) reductase catalysis

Mycobacterium tuberculosis adenosine 5'-phosphosulfate reductase (APR) catalyzes the first committed step in sulfate reduction for the biosynthesis of cysteine and is essential for survival in the latent phase of tuberculosis infection. The reaction catalyzed by APR involves the nucleophilic attack by conserved Cys-249 on adenosine 5'-phosphosulfate, resulting in a covalent S-sulfocysteine intermediate that is reduced in subsequent steps by thioredoxin to yield the sulfite product. Cys-249 resides on a mobile active site lid at the C terminus, within a K(R/T)ECG(L/I)H motif. Owing to its strict conservation among sulfonucleotide reductases and its proximity to the active site cysteine, it has been suggested that His-252 plays a key role in APR catalysis, specifically as a general base to deprotonate Cys-249. Using site-directed mutagenesis, we have changed His-252 to an alanine residue and analyzed the effect of this mutation on the kinetic parameters, pH rate profile, and ionization of Cys-249 of APR. Interestingly, our data demonstrate that His-252 does not perturb the pK(a) of Cys-249 or play a direct role in rate-limiting chemical steps of the reaction. Rather, we show that His-252 enhances substrate affinity via interaction with the α-phosphate and the endocyclic ribose oxygen. These findings were further supported by isothermal titration calorimetry to provide a thermodynamic profile of ligand-protein interactions. From an applied standpoint, our study suggests that small-molecules targeting residues in the dynamic C-terminal segment, particularly His-252, may lead to inhibitors with improved binding affinity.     

Estimating the prevalence and strength of non-independent predator effects

Understanding whether multiple predator species have independent effects on shared prey is critical for understanding community dynamics. We describe the prevalence and strength of non-independence between predators by quantifying the prey’s risk of predation and the degree to which it deviates from the risk predicted from a null model of independent predator effects. Specifically, we document how frequently non-independent effects occur among ten different multiple predator combinations with mayfly larvae as prey. These predator combinations vary both predator density and predator species richness. Overall, the predator effects were non-independent and translated to an average of 27% fewer prey being consumed compared to independent predator effects. Non-independence of this magnitude is likely to have population level consequences for the prey and influence the distribution or prey preference of predators. Closer inspection shows that much of the risk reduction in this system is weak, to the point of being indistinguishable from independent predator effects, while few effects are strong. This pattern of many weak interactions and few strong ones parallels the pattern of interaction strengths documented previously in intertidal communities. Consequently, understanding strong interactors in multiple predator systems may help us understand the importance of a species.     

Effect of circular permutation on the structure and function of type 1 blue copper center in azurin

Type 1 copper (T1Cu) proteins are electron transfer (ET) proteins involved in many important biological processes. While the effects of changing primary and secondary coordination spheres in the T1Cu ET function have been extensively studied, few report has explored the effect of the overall protein structural perturbation on active site configuration or reduction potential of the protein, even though the protein scaffold has been proposed to play a critical role in enforcing the entatic or “rack‐induced” state for ET functions. We herein report circular permutation of azurin by linking the N‐ and C‐termini and creating new termini in the loops between 1^st and 2^nd β strands or between 3^rd and 4^th β strands. Characterization by electronic absorption, electron paramagnetic spectroscopies, as well as crystallography and cyclic voltammetry revealed that, while the overall structure and the primary coordination sphere of the circular permutated azurins remain the same as those of native azurin, their reduction potentials increased by 18 and 124 mV over that of WTAz. Such increases in reduction potentials can be attributed to subtle differences in the hydrogen‐bonding network in secondary coordination sphere around the T1Cu center.     

城市绿地的生态环境效应研究进展

工业化的快速发展和城市化进程的加剧,使得城市生态环境的破坏和污染愈来愈严重,加速了城市的“生态环境危机”.城市绿地能够通过植被的光合作用、吸收作用、隔离阻挡作用、蒸腾以及蒸散作用改善城市的生态环境.目前研究主要集中在降温、增湿、固碳释氧、降噪、抗污染、生物多样性保护等6个方面.关于城区绿地对6种生态环境要素的改善程度、影响范围、机理问题以及不同植被种类生态环境效应的差异情况等,国内外学者都进行了大量详细的研究.基于传统实地观测数据进行城市绿地的生态环境效应研究是目前主要使用的研究方法,部分学者开始使用遥感技术手段研究城区绿地的降温增湿效应.在总结和分析前人研究成果的基础上,从研究内容的不同着手,主要总结并系统评估了目前关于城市绿地6种生态环境效应的研究,为今后相关研究提供参考.