Classification of a Haemophilus influenzae ABC Transporter HI1470/71 through Its Cognate Molybdate Periplasmic Binding Protein, MoIA

molA (HI1472) from H. influenzae encodes a periplasmic binding protein (PBP) that delivers substrate to the ABC transporter MolB_2C_2 (formerly HI1470/71). The structures of MolA with molybdate and tungstate in the binding pocket were solved to 1.6 and 1.7 Å resolution, respectively. The MolA-binding protein binds molybdate and tungstate, but not other oxyanions such as sulfate and phosphate, making it the first class III molybdate-binding protein structurally solved. The ~100 μM binding affinity for tungstate and molybdate is significantly lower than observed for the class II ModA molybdate-binding proteins that have nanomolar to low micromolar affinity for molybdate. The presence of two molybdate loci in H. influenzae suggests multiple transport systems for one substrate, with molABC constituting a low-affinity molybdate locus

Silicate determination in sea water: toward a reagentless electrochemical method

ilicate has been determined in sea water by four different electrochemical methods based on the detection of the silicomolybdic complex formed in acidic media by the reaction between silicate and molybdenum salts. The first two methods are based on the addition of molybdate and protons in a seawater sample in an electrochemical cell. Cyclic voltammetry presents two reduction and two oxidation peaks giving four values of the concentration and therefore increasing the precision. Then chronoamperometry is performed on an electrode held at a constant potential. A semi-autonomous method has been developed based on the electrochemical anodic oxidation of molybdenum, the complexation of the oxidation product with silicate and the detection of the complex by cyclic voltammetry. This method is tested and compared with the classical colorimetric one during ANT XXIII/3 cruise across Drake Passage (January–February 2006). The detection limit is 1 μM and the deviation between both methods is less than 3% for concentrations higher than 10 μM. Finally a complete reagentless method with a precision of 2.6% is described based on the simultaneous formation of the molybdenum salt and protons in a divided electrochemical cell. This latter method should be very useful for developing a reagentless sensor suitable for long term in situ deployments on oceanic biogeochemical observatories

Phosphate Contaminant Detection in Water Through a Paper-based Microfluidic Device

This report describes a project aimed at developing a low-cost, portable, on-site, user-friendly system for detecting different concentrations of phosphate in drinking water. Phosphate is a natural chemical, but toxic in large concentrations; detection is therefore important to avoid drinking contaminated water. Despite this fact, no cheap, and/or nontoxic system for phosphate detection is yet on the market. The detection system utilizes a paper-based microfluidic device to automate the electrochemical detection process, which normally requires expert use of lab equipment. When combined with a portable potentiostat that works with a mobile app, the device will allow untrained users to determine if any source of drinking water contains unsafe levels of phosphate without equipment or training, and to communicate that information to a central database for further analysis. Those of any background, particularly in developing countries, will be able to maintain health and raise awareness about clean water. Microfluidic devices are useful tools for the detection of water contaminants, but there is a gap in technology for the detection of phosphate. Our phosphate detection system is a paper-based microfluidic device with an already-developed voltammetry device that automates the detection process so that any user can safely find phosphate in water. The system will provide a binary analysis about whether the water is safe to consume or not. Completion of the project provides a valuable tool to both average customers in developing countries and scientific researchers in determining the safety of drinking water

Precipitation of molybdenum from homogeneous solution by alpha-benzoinoxime

Thesis (M.A.)--Boston UniversityThere are two possible approaches to the precipitation of molybdenum from homogeneous solution by benzoinoxime. One involves the preparation of a derivative of benzoin which can be converted under either neutral or alkaline conditions to benzoinoxime. If free molybdenum is present in solution, complexation and deposition of the molybdenum-benzoinoxime complex will take place. The other approach involves the preparation of benzoinoxime in the presence of molybdenum. As soon as sufficient quantities of the complexing agent have been generated molybdenumbenzoinoxime complex deposition occurs. The former path was attempted first through synthesis of methylbensoinether and methylbenzoinetheraxime in hopes that the latter material could be converted to benzoinoxime under conditions which are to exist in a later precipitation procedure. It was found, however, tbat the methylbenzoinoxime does not deccmpose either under neutral or basic conditions. Under acidic conditions (which we do not want to use since an acidic procedure for the direct quantitative precipitation ot molybdenum by alpha-benzoinoxime is already on record) the methylbenzoinetheroxime is found to decompose into its components -- methylbensoinether and hydroxylamine [TRUNCATED

Novel Inducers of the Envelope Stress Response BaeSR in Salmonella Typhimurium: BaeR Is Critically Required for Tungstate Waste Disposal

The RpoE and CpxR regulated envelope stress responses are extremely important for SalmonellaTyphimurium to cause infection in a range of hosts. Until now the role for BaeSR in both the Salmonella Typhimurium response to stress and its contribution to infection have not been fully elucidated. Here we demonstrate stationary phase growth, iron and sodium tungstate as novel inducers of the BaeRregulon, with BaeR critically required for Salmonella resistance to sodium tungstate. We show that functional overlap between the resistance nodulation-cell division (RND) multidrug transporters, MdtA, AcrD and AcrB exists for the waste disposal of tungstate from the cell. We also point to a role for enterobactinsiderophores in the protection of enteric organisms from tungstate, akin to the scenario in nitrogen fixing bacteria. Surprisingly, BaeR is the first envelope stress response pathway investigated in S. Typhimurium that is not required for murine typhoid in either ityS or ityR mouse backgrounds. BaeR is therefore either required for survival in larger mammals such as pigs or calves, an avian host such as chickens, or survival out with the host altogether where Salmonella and related enterics must survive in soil and water

Ultratrace determination of phosphorus in ultrapurified water by a slope comparison method

The analytical method for the determination of phosphorus in ultrapurified water was developed. Ultrapurified water was evaporated to concentrate phosphorus and the final sample volume for analysis was 10 ml. In 0.55 mol 1(-1) HCl, orthophosphate forms molybdophosphate, and then the molybdophosphate forms ion associate with Malachite Green (MG), which can be collected on a tiny membrane filter (diameter: 5 mm, and effective filtering diameter: 2 mm). After the ion associate on the membrane filter is dissolved together with the membrane filter in I ml of methyl cellosolve (MC), the absorbance of MC solution is measured at 627 nm by a flow injection-spectrophotometric detection technique. When 10 ml of the sample solution was used for the procedures and absorbance measurement, the calibration graph is linear up to about 500 ng 1(-1) of phosphorus and the detection limit was 8 ng 1(-1) (S/N = 3). For the determination of phosphorus in an ultrapurified water, 10-40 ml of sample solutions were transferred into poly(tetrafluoroethylene) (PTFE) beaker and evaporated to 5 ml or to dryness. To them, 0.003 mol 1(-1) HCl was added to get 10 ml of final solution, which was used as sample. Phosphate is determined by comparing the slope of the varied sample volume after evaporation/concentration with a slope of the standard calibration graph (a slope comparison method: SCM). The SCM enables to evaluate the concentration of phosphate in ultrapurified waters more sensitively and accurately

Thermochemistry of iron manganese oxide spinels

Oxide melt solution calorimetry has been performed on iron manganese oxide spinels prepared at high temperature. The enthalpy of formation of (MnxFe1−x)3O4 at 298 K from the oxides, tetragonal Mn3O4 (hausmannite) and cubic Fe3O4 (magnetite), is negative from x=0 to x=0.67 and becomes slightly positive for 0.670.6) spinels of intermediate compositions. The enthalpies of formation are discussed in terms of three factors: oxidation–reduction relative to the end-members, cation distribution, and tetragonality. A combination of measured enthalpies and Gibbs free energies of formation in the literature provides entropies of mixing. ΔSmix, consistent with a cation distribution in which all trivalent manganese is octahedral and all other ions are randomly distributed for x>0.5, but the entropy of mixing appears to be smaller than these predicted values for x<0.4

Transformation of 1,1,1-trichloroethane in an anaerobic packed-bed reactor at various concentrations of 1,1,1-trichloroethane, acetate and sulfate

Biotransformation of 1,1,1-trichloroethane (CH3CCl3) was observed in an anaerobic packed-bed reactor under conditions of both sulfate reduction and methanogenesis. Acetate (1 mM) served as an electron donor. CH3CCl3 was completely converted up to the highest investigated concentration of 10 µM. 1,1-Dichloroethane and chloroethane were found to be the main transformation products. A fraction of the CH3CCl3 was completely dechlorinated via an unknown pathway. The rate of transformation and the transformation products formed depended on the concentrations of CH3CCl3, acetate and sulfate. With an increase in sulfate and CH3CCl3 concentrations and a decrease in acetate concentration, the degree of CH3CCl3 dechlorination decreased. Both packed-bed reactor studies and batch experiments with bromoethanesulfonic acid, an inhibitor of methanogenesis, demonstrated the involvement of methanogens in CH3CCl3 transformation. Batch experiments with molybdate showed that sulfate-reducing bacteria in the packed-bed reactor were also able to transform CH3CCl3. However, packed-bed reactor experiments indicated that sulfate reducers only had a minor contribution to the overall transformation in the packed-bed reactor.