Catalytic defects in mutants of class II histidyl-tRNA synthetase from Salmonella typhimurium previously linked to decreased control of histidine biosynthesis regulation

The expression of histidine biosynthetic genes in enteric bacteria is regulated by an attenuation mechanism in which the level of histidyl-tRNA serves as a key sensor of the intracellular histidine pool. Among the early observations that led to the formation of this model for Salmonella typhimurium were the identification of mutants in the gene (hisS) encoding histidyl-tRNA synthetase. We report here the detailed biochemical characterization of five of these S. typhimurium bradytrophic mutants isolated by selection for resistance to histidine analogs, including identification of the deduced amino acid substitutions and determination of the resulting effects on the kinetics of adenylation and aminoacylation. Using the crystal structure of the closely related Escherichia coli histidyl-tRNA synthetase (HisRS) as a guide, two mutants were mapped to a highly conserved proline residue in motif 2 (P117S, P117Q), and were correlated with a fivefold decrease in the kcat for the pyrophosphate exchange reaction, as well as a tenfold increase in the Km for tRNA in the aminoacylation reaction. Another mutant substitution (A302T) mapped to a residue adjacent to the histidine binding pocket, leading to a tenfold increase in Km for histidine in the pyrophosphate exchange reaction. The remaining two mutants (S167F, N254T) substitute residues in or directly adjacent to the hinge region, which joins the insertion domain between motif 2 and motif 3 to the catalytic core, and cause the Km for tRNA to increase four- to tenfold. The kinetic analysis of these mutants establishes a direct link between critical interactions within the active site of HisRS and regulation of histidine biosynthesis, and provides further evidence for the importance of local conformational changes during the catalytic cycle.     

Novel potentiometric sensors for the selective determination of domperidone

The fabrication and electrochemical response characteristics of two novel potentiometric sensors for the selective determination of domperidone (DOM) are described. The two fabricated sensors incorporate DOM–PTA (phosphotungstic acid) ion pair as the electroactive material. The sensors include a PVC membrane sensor and a carbon paste sensor. The sensors showed a linear, stable, and near Nernstian slope of 56.5 and 57.8 mV/decade for PVC membrane and carbon paste sensors, respectively over a relatively wide range of DOM concentration (1.0 × 10^?1–1.0 × 10^?5 and 1.0 × 10^?1–3.55 × 10^?6 M). The response time of DOM–PTA membrane sensor was less than 25 s and that in the case of carbon paste sensor was less than 20 s. A useful pH range of 4–6 was obtained for both types of sensors. A detection limit of 7.36 × 10^?5 M was obtained for PVC membrane sensor and 1.0 × 10^?6 M was obtained for carbon paste sensor. The proposed sensors showed very good selectivity to DOM in the presence of a large number of other interfering ions. The analytical application of the developed sensors in the determination of the drug in pharmaceutical formulations such as tablets was investigated. The results obtained are in good agreement with the values obtained by the standard method. The sensors were also applied for the determination of DOM in real samples such as urine by the standard addition method.     

Selective Conversion of Glycerol to Propylene Glycol Over Fixed Bed Raney® Cu Catalysts

Net propylene glycol (1,2 propanediol) yields of up to 94% at 100% glycerol conversion have been achieved over a fixed bed Raney^® Cu catalyst in trickle bed mode, at relatively low total pressure, 14 bar (200 psig), and minimal feedstock dilution (20 wt% water). The main identified byproducts are ethylene glycol and ethanol (each <2%), with methanol and 1,3 diol both <1%. The other key operating parameters for high yields are a narrow optimum in temperature (near 205 °C), and a high H₂/liquid flow ratio, about 375/0.05 by volume. The effects of chromium promotion have also been studied for effects on side reactions and rates. Our evidence points to initial dehydrogenation as the rate-limiting step in a likely three step mechanism.     

Agro-ecological nitrogen management in soils vulnerable to nitrate leaching: a case study in the Lower Suwannee Watershed

Environmental benefits associated with reduced rates of nitrogen (N) application, while maintaining economically optimum yields have economic and social benefits. Although N is an indispensable plant nutrient, residual soil N could leach out to contaminate groundwater and surface water resources, particularly in sandy soils. A 2-year field study was conducted in an established bermudagrass (Cynodon dactylon) pasture in the Lower Suwannee Watershed, Florida, to evaluate N application rates on forage yield, forage quality, and nitrate (NO₃-N) leaching in rapidly permeable upland sandy soils. Four N application rates (30, 50, 70, and 90 kg N ha⁻¹ harvest⁻¹) corresponding to 0.33, 0.55, 0.77 and IX, respectively, of recommended N rate (90 kg N ha⁻¹ harvest⁻¹) for bermudagrass hay production in Florida were evaluated vis-à-vis an unfertilized (0 N) control. Suction cups were installed near the center of each plot at two depths (30 and 100 cm) to monitor NO₃-N leaching. The grass was harvested at 28 days intervals to determine dry matter yield, N uptake, and herbage nutritive value. Nitrogen application at the recommended rate produced the greatest total dry matter yield (~18.4 Mg ha⁻¹ year⁻¹), but a modeled economically optimum N rate of ~57 kg N ha⁻¹ harvest⁻¹ (~60% of the recommended N rate) projected an average dry matter yield of ~17.3 Mg ha⁻¹ year⁻¹, which represents >90% of the observed maximum yield. Nitrogen application increased nutritive quality of the grass, but increases in N application rate above 30 kg N ha⁻¹ did not result in significant increases in in vitro digestible organic matter concentration, and tissue crude protein was not significant above 50 kg N ha⁻¹. Across the sampling period, treatments with N rates ≤50 kg N ha⁻¹ harvest⁻¹ had leachate NO₃-N concentration below the maximum contaminant limit of <10 mg l⁻¹. Conversely, applying N at rates ≥70 kg N ha⁻¹ harvest⁻¹ resulted in leachate N concentration that exceeded the maximum contaminant limit, and suggest high risk of impacting groundwater quality, if such rates are applied to soils with coarse (sand) textures. The study demonstrates that recommendation of a single N application rate may not be appropriate under all agro-climatic conditions and, thus, a site-specific evaluation of best N management strategy is critical.     

Recovered oil from palm-pressed fiber: A good source of natural carotenoids,vitamin E,and sterols

Recovered fiber from pressed palm fruits, which is normally burned as fuel to provide energy for the palm oil mills, has now been found to be a rich source of carotenoids, vitamin E (tocopherol and tocotrienols), and sterols. Residual oil (5–6% on dry basis) extracted from palm press fibers contains a significant quantity of carotenoids (4000–6000 ppm), vitamin E (2400–3500 ppm), and sterols (4500–8500 ppm). The major identified carotenoids are α-carotene (19.5%), β-carotene (31.0%), lycopene (14.1%), and phytoene (11.9%). In terms of vitamin E, α-tocopherol constitutes about 61% of the total vitamin E present, the rest being tocotrienols (α-, γ-, and δ-). The major sterols present are β-sitosterol (47%), campesterol (24%), and stigmasterol (15%). The oil extracted from palm-pressed fiber is contaminated with about 30% of palm kernel oil. The quality of this fiber oil is slightly lower than that of crude palm oil in terms of the content of free fatty acids, peroxide value, and anisidine value.     

One Pot Synthesis and Characterization of Cesium Doped SnO2 Nanocrystals via a Hydrothermal Process

The physico-chemical properties of cesium doped SnO2 nanocrystals synthesized by wet chemical method have been investigated. Scanning electron microscopy (SEM), X-ray diffraction (XRD), energy dispersive X-ray analysis (EDAX), inductively coupled plasma (ICP), atomic absorption spectroscopic (AAS) analyses, UV-vis-NIR spectral studies and dielectric studies were carried out for both pure SnO2 and cesium doped SnO2 nano-samples. All samples of SnO2 did not show any metallic cluster, but the sample containing cesium as a dopant displayed significant activity. The products formed were chloride and water representing a competitive advantage from the stand point of environmental protection.