Role of cis-cis muconic acid in the catalysis of Pseudomonas putida chlorocatechol 1,2-dioxygenase

Chlorocatechol 1,2-dioxygenase (1,2-CCD) is a non-heme iron protein involved in the intradiol cleavage of aromatic compounds that are recalcitrant to biodegradation. In particular, 1,2-CCD catalyzes the conversion of catechol and its halogenated derivatives to cis-cis muconic acid. In this study we describe a series of experiments concerning the interaction of chlorocatechol 1,2-dioxygenase from Pseudomonas putida (Pp1,2-CCD) with cis-cis muconic acid. We used single-injection ITC to show that the reaction product inhibits enzyme kinetics. DSC and EPR measurements probed whether this was accomplished by a direct binding of the product to the enzyme active site. DSC shows that cis-cis muconic acid affects the thermal unfolding of the protein and allowed us to estimate a binding constant. Furthermore, EPR spectra of the Fe(III) center demonstrate that, upon product binding, a significant decrease in resonance intensity is observed, indicating that cis-cis muconic acid binds directly to the active site. Based on the increasing interest for understanding dioxygenases mechanism of action and, moreover, how to control such process, our data indicate that the product of the reaction does play a relevant role in the catalysis and should therefore be taken into account when one thinks about ways of regulating enzyme activity. (C) 2010 Elsevier B.V. All rights reserved.

Useful Strategies for Algal Volatile Analysis

The production of volatile organic compounds (VOC) by plants is well known. However, few scientific groups have studied VOC produced by green, brown and red algae. Headspace collection of volatiles and solid phase microextraction, as well as the traditional extraction by hydrodistillation combined with analytical chromatographic techniques (i.e., GC-MS), have significantly improved the investigation of VOC from plants and algae. The major volatile compounds found in seaweeds are hydrocarbons, terpenes, phenols, alcohols, aldehydes, ketones, esters, fatty acids and halogen or sulfur-containing compounds. This article presents an overview of VOC isolated from and identified in marine macro-algae. Focus is given to non-halogenated and non-sulfur volatile compounds, as well as strategies to analyze and identify algal VOC by GC-MS.

Atmospheric pressure photoionization mass spectrometry as a tool for the investigation of the hydrolysis reaction mechanisms of phosphite antioxidants

The hydrolysis reaction mechanism of phosphite antioxidants is investigated by liquid chromatography-mass spectrometry (LC/MS). The phosphites were chosen because they differed in chemical structure and phosphorus content. Dopant assisted-atmospheric pressure photoionization (DA-APPI) is chosen as the ion source for (lie ionization of the compounds. [it our previous work, DA-APPI was shown to offer an attractive alternative to atmospheric pressure chemical ionization (APCI) since it provided background-ion free mass spectra and higher sensitivity [M. Papanastasiou, et al., Polymer Degradation and Stability 91 (11) (2006) 2675-2682]. In positive ion mode, the molecules are generally detected in their protonated form. In negative ion mode, the phosphites are unstable and only fragment ions are observed: these however, are characteristic of each phosphite and may be used for the identification of the analytes in complex mixtures. The analytes under investigation are exposed to accelerated humid ageing conditions and their hydrolytic pathway and stability is investigated. Different substituents around the phosphorus atom are shown to have a significant effect on the stability of the phosphites, with phenol substituents producing very hydrolytically stable structures. Alkanox P24 and PEP-36 follow a similar hydrolytic pathway via the scission of the first and then the second P-O-phenol bonds, eventually leading to the formation of phenol, Phosphorous acid and pentaerythritol as end products. HP-10 exhibits a rather different Structure and the products detected suggest scission of either the P-O-hydrocarbon or one of the P-O-phenol bonds. A phenomenon similar to that of autocatalysis is observed for all phosphites and is attributed to the formation of dialkyl phosphites as intermediate products. (C) 2008 Elsevier B.V. All rights reserved.

The effects of hydrogen sulfide on the polymer electrolyte membrane fuel cell anode catalyst: H(2)S-Pt/C interaction products

The performance of a polymer electrolyte membrane fuel cell (PEMFC) operating on a simulated hydrocarbon reformate is described. The anode feed stream consisted of 80% H(2),similar to 20% N(2), and 8 ppm hydrogen sulfide (H(2)S). Cell performance losses are calculated by evaluating cell potential reduction due to H(2)S contamination through lifetime tests. It is found that potential, or power, loss under this condition is a result of platinum surface contamination with elemental sulfur. Electrochemical mass spectroscopy (EMS) and electrochemical techniques are employed, in order to show that elemental sulfur is adsorbed onto platinum, and that sulfur dioxide is one of the oxidation products. Moreover, it is demonstrated that a possible approach for mitigating H(2)S poisoning on the PEMFC anode catalyst is to inject low levels of air into the H(2)S-contaminated anode feeding stream. (C) 2011 Elsevier B.V. All rights reserved.

Emissions Characterisation of residential pellet boilers during start-up and stop periods

In this study, gaseous emissions and particles are measured during start-up and stop periods for an over-fed boiler and an under-fed boiler. Both gaseous and particulate matter emissions are continuously measured in the laboratory. The measurement of gaseous emissions includes oxygen (O2), carbon dioxide (CO2), carbon monoxide (CO), nitrogen oxide and (NO). The emissions rates are calculated from measured emissions concentrations and flue gas flow. The behaviours of the boilers during start-up and stop periods are analysed and the emissions are characterised in terms of CO, NO, TOC and particles (PM2.5 mass and number). The duration of the characterised periods vary between two boilers due to the difference in type of ignition and combustion control. The under-fed boiler B produces higher emissions during start-up periods than the over-fed boiler A. More hydrocarbon and particles are emitted by the under-fed boiler during stop periods. Accumulated mass of CO and TOC during start-up and stop periods contribute a major portion of the total mass emitted during whole operation. However, accumulated mass of NO and PM during start-up and stop periods are not significant as the duration of emission peak is relatively short.

Emissions from realistic operation of residential wood pellets heating systems

Emissions from residential combustion appliances vary significantly depending on the firing behaviours and combustion conditions, in addition to combustion technologies and fuel quality. Although wood pellet combustion in residential heating boilers is efficient, the combustion conditions during start-up and stop phases are not optimal and produce significantly high emissions such as carbon monoxide and hydrocarbon from incomplete combustion. The emissions from the start-up and stop phases of the pellet boilers are not fully taken into account in test methods for ecolabels which primarily focus on emissions during operation on full load and part load. The objective of the thesis is to investigate the emission characteristics during realistic operation of residential wood pellet boilers in order to identify when the major part of the annual emissions occur. Emissions from four residential wood pellet boilers were measured and characterized for three operating phases (start-up, steady and stop). Emissions from realistic operation of combined solar and wood pellet heating systems was continuously measured to investigate the influence of start-up and stop phases on total annual emissions. Measured emission data from the pellet devices were used to build an emission model to predict the annual emission factors from the dynamic operation of the heating system using the simulation software TRNSYS. Start-up emissions are found to vary with ignition type, supply of air and fuel, and time to complete the phase. Stop emissions are influenced by fan operation characteristics and the cleaning routine. Start-up and stop phases under realistic operation conditions contribute 80 – 95% of annual carbon monoxide (CO) emission, 60 – 90% total hydrocarbon (TOC), 10 – 20% of nitrogen oxides (NO), and 30 – 40% particles emissions. Annual emission factors from realistic operation of tested residential heating system with a top fed wood pelt boiler can be between 190 and 400 mg/MJ for the CO emissions, between 60 and 95 mg/MJ for the NO, between 6 and 25 mg/MJ for the TOC, between 30 and 116 mg/MJ for the particulate matter and between 2x10-13 /MJ and 4x10-13 /MJ for the number of particles. If the boiler has the cleaning sequence with compressed air such as in boiler B2, annual CO emission factor can be up to 550 mg/MJ. Average CO, TOC and particles emissions under realistic annual condition were greater than the limits values of two eco labels. These results highlight the importance of start-up and stop phases in annual emission factors (especially CO and TOC). Since a large or dominating part of the annual emissions in real operation arise from the start-up and stop sequences, test methods required by the ecolabels should take these emissions into account. In this way it will encourage the boiler manufacturers to minimize annual emissions. The annual emissions of residential pellet heating system can be reduced by optimizing the number of start-ups of the pellet boiler. It is possible to reduce up to 85% of the number of start-ups by optimizing the system design and its controller such as switching of the boiler pump after it stops, using two temperature sensors for boiler ON/OFF control, optimizing of the positions of the connections to the storage tank, increasing the mixing valve temperature in the boiler circuit and decreasing the pump flow rate. For 85 % reduction of start-ups, 75 % of CO and TOC emission factors were reduced while 13% increase in NO and 15 % increase in particle emissions was observed.

Baseline assessment of metals and hydrocarbons in the sediments of Lake Mulwala, Australia

An assessment of hydrocarbon and metal/metalloids (arsenic, cadmium, copper, lead, mercury, selenium, zinc) contamination in sediments from Lake Mulwala, Australia, was undertaken. The objectives of the study were: (i) to determine the extent of contamination in the lake sediments, compared to Australian and international sediment quality guidelines, and (ii) to attempt to identify the contaminant sources to the lake. With the exception of a few samples containing elevated levels of arsenic and/or mercury, the levels of all contaminants in the sediment samples taken from the lake were below the 'lower trigger' of the Australian Sediment Quality Guidelines that would warrant further investigation. High molecular weight hydrocarbons (up to 700 mg kg−1) were found in most sediment samples. Non-metric statistical analysis indicated that the contaminant distribution was different in different parts of the lake, with the lowest concentrations generally found at the influent to the lake. No definitive source(s) of contamination could be identified for either metalloids or hydrocarbons.

Late Paleozoic depositional history of the Tarim basin, northwest China: an integration of biostratigraphic and lithostratigraphic constraints

This study provides the first detailed lithostratigraphic and biostratigraphic constraints for improving stratigraphic resolution for hydrocarbon prospecting and exploration in the Tarim basin. A total of 49 stratigraphic units (38 formations and 11 members), ranging in age from the latest Devonian to Permian, are reviewed or redefined in terms of nomenclatures, lithology, age constraints, and lateral distributions based on the detailed field works or newly published data. Of these, the Piqiang Formation (new formation) is proposed to include the reefal carbonates of Asselian-Sakmarian age from the northern Tarim. The subsurface upper Paleozoic stratigraphic framework of the desert areas of the basin is also established for the first time. The high-resolution, basinwide stratigraphic correlations reveal that the sedimentation of the basin in the late Paleozoic was extremely uneven. Of these, the Famennian to Changhsingian successions are completely recorded in the south-western margin areas of the basin. Here, five eustatic sedimentary cycles are well recognizable, suggesting the sedimentation was more eustatically controlled and little affected by local tectonism. The late Paleozoic successions of both Kalpin and Taklimakan regions are commonly interrupted by major hiatuses at various horizons, suggesting that the sedimentation was apparently modified by local tectonism. Of these, the northward movement of the Tarim block and its subsequent collision with the Yili microcontinent (part of the Kazakhstan plate) may be principally accountable for the discrepancy in the sedimentation of the various regions in the basin in the late Paleozoic.

Synthesis of 2-oxo-16-(3', 4'-methylenedioxyphenyl)-trans-15-hexadecene

This work presents the progress made towards synthesizing 2-oxo-16-(3', 4'methylenedioxyphenyl)-trans-15-hexadecene, an antimycobacterial compound that was originally isolated from the leaves of Piper Sanctum. The hydrocarbon chain of the molecule was synthesized first by opening a 15-pentadecanolactone ring by means of HI, and performing an E2 elimination reaction on the molecule followed by an organolithium reaction with CH3Li. Hexadec-15-en-2-one that was afforded this way was later reacted with 5-bromobenzo[d][1,3]dioxole following the appropriate Heck reaction protocol that allows for the formation of a palladium catalyzed carbon-carbon bond. The modes of action of 2-oxo-16-(3', 4'-methylenedioxyphenyl)-trans-15hexadecene are comparable to the ones of rifampicin, a marketable drug that has been successfully used in the treatment of tuberculosis in the past. Additionally, this compound can serve as an intermediate towards the synthesis of 2-oxo-16-(3', 4' methylenedioxyphenyl)-hexadecane and 2-oxo-14-(3', 4' -methylenedioxyphenyl) tetradecane, both strong inhibitors of the growth of Mycobacterium tuberculosis. Lastly, due to Multi-Drug Resistant tuberculosis, there has been an increasing need to find alternative cures for tuberculosis. Therefore, the work on 2-qxo-16-(3', 4'methylenedioxyphenyl)-trans-15-hexadecene is not only chemically interesting but it is also biologically important.

Androgenic activity of effluent from forty-five municipal waste water treatment plants in Victoria, Australia

It is well known that waste water treatment plant (WWTP) effluents are estrogenic. There has been much less consideration of the androgenic activity of WWTP effluents. To partly address the shortage of information on androgens in Australian WWTP effluents, in August 2006, and again in 2007, we collected discharges from up to 45 Victorian WWTPs (~25% of all WWTPs in Victoria), grouped by treatment process, i.e. activated sludge, extended aeration, and lagoon based treatment, and measured the total estrogenic, androgenic, retinoic acid, and aromatic hydrocarbon hydrogenase activity of the effluents using a hybrid yeast bioassay. This paper will concentrate on the androgenic activity and male hormone concentrations.

Organotin-oxo clusters

This thesis reports on the development and expansion of reliable synthetic di-and multi-tin precursors for the assembly of oligomeric organotin-oxo compounds in which the shape, dimension and tin nuclearity can be controlled. The reaction of polymeric diorganotin oxides, (R2SnO)m (R = Me, Et, n-Bu, n-Oct, c-Hex, i-Pr, Ph), with saturated aqueous NH4X solutions (X = F, Cl, Br, I, OAc) in refluxing 1,4-dioxane afforded in high yields dimeric tetraorganodistannoxanes, [R2(X)SnOSn(X)R2]2, and in a few cases diorganotin dihalides or diacetates, R2SnX2. This method appears to be particularly good for the synthesis of halogenated tetraorganodistannoxanes but a less suitable method for the preparation of dicarboxylato tetraorganodistannoxanes. Identification of [R2(OH)SnOSn(X)R2]2 (R = n-Bu; X = Cl, Br) and [R2(OH)SnOSn(X)R2][R2(X)SnOSn(X)R2] suggest a serial substitution mechanism starting from [R2(OH)SnOSn(OH)R2]2. A series of α, ω -bis(triphenylstannyl)alkanes, [Ph3Sn]2(CH2)n (n = 3-8, 10, 12) and some of their derivatives were synthesised and characterised. These α, ω-bis(triphenylstannyl)alkanes, [Ph3Sn]2(CH2)n were converted to the corresponding halides [R(Cl)2Sn]2(CH2)n (R = CH2SiMe3) and subsequently to the polymeric oxides {[R(0)Sn]2(CH2)n}m. Reaction of {[R(O)Sn]2(CH2)n}m with [R(Cl)2Sn]2(CH2)n. (n = 3, n' = 4 and n = 4, n' = 3) in toluene at 100°C results in a mixture of symmetric and asymmetric double ladders, where different spacer chain lengths (n and n') provide the source of asymmetry. The coexistence at high temperature of separate 119Sn NMR signals belonging to symmetric and asymmetric double ladders suggests an equilibrium that is slow on the 119Sn NMR time scale and the position of which is temperature dependent. However, 119Sn NMR spectroscopic experiments of {[R(0)Sn]2(CH2)3}m with [R(Cl)2Sn]2(CH2)n for longer spacers (n - 5, 6, 8, 10, 12) reveal that molecular self-assembly of symmetric spacer-bridged di-tin precursors of equal chain length is preferred over asymmetric species. An ether-bridged di-tin tetrachloride [R(Cl)2Sn(CH2)3]2O (R = CH2SiMe3) and its corresponding polymeric oxide {[R(O)Sn(CH2)3]2O}m were synthesised and characterised. Reaction of [R(Cl)2Sn(CH2)3]2O with {[R(O)Sn(CH2)3]2O}m results in a unique functionalised double ladder {{[RSn(Cl)](CH2)3O(CH2)3[RSn(Cl)]}O}4 whose structure in the solid state was determined by X-ray analysis. Identification of tetrameric functionalised double ladder as well as dimeric and monomeric species suggest the existence of an equilibrium in solution. The feasibility of the functionalised double ladder to form host-guest complexes with a variety of metal cations is investigated using electrospray mass spectrometry (ESMS). Evidence for such complexes is found only for sodium cations. The reaction between {[R(O)Sn]2(CH2)n}m (n = 3, 4, 8, 10) and triflic acid is described. The initial formed products [RSn(CH2)nSnR](OTf)4 are easily hydrolysed. For n = 3, self-assembly leads to a discrete double ladder type structure, {{[RSn(OH)](CH2)3[RSn(H2O)]}O}44OTf, which is the first example of a cationic double ladder. For n ≥ 3, hydrolysis gives polymeric products, as demonstrated by the crystal structure of {[(H2O)(OH)RSn]2(CH2)4-2OTf2H2O}m. Two spacer-bridged terra-tin octachlorides [R(Cl)2Sn(CH2)3Sn(Cl)2]2(CH2)n (R = CH2SiMes; n = 1, 8) and their corresponding polymeric oxides {[R(O)Sn(CH2)3Sn(O)]2(CH2)n}m were successfully synthesised and characterised. Attempts were made to synthesise quadruple ladders from these precursors. Reactions of [R(Cl)2Sn(CH2)3Sn(Cl)2]2CH2 with {[R(O)Sn(CH2)3Sn(O)]2CH2}m or (Y-Bu2SnO)3 result in, mostly insoluble, amorphous solids. Reactions of [R(Cl)2Sn(CH2)3Sn(Cl)2]2(CH2)8 with {[R(O)Sn(CH2)3Sn(O)]2(CH2)8}m or (t-Bu2SnO)s result in new tin-containing species which are presumably oligomeric. The synthesis of a series of alkyl-bridged di-tin hexacarboxylates [(RCO2)3Sn]2(CH2)n (n = 3, 4; R = Ph, c-C6H11, CH3, C1CH2) is also reported. The hydrolysis of these compounds is facile and complex. There appears to be no correlation between spacer chain length and hydrolysis product. However, the conjugate acid strength of the carboxylate does appear to be important. In general only insoluble amorphous polymeric organotin-oxo compounds were obtained.

Tribology of lubricated nitrocarburised and titanium carbonitride surfaces

In the current work, two different coatings, nitrocarburised (CN) and titanium carbonitride (TiCN) on M2 grade high speed tool steel, were prepared by commercial diffusion and physical vapour deposition (PVD) techniques, respectively. Properties of the coating were characterised using a variety of techniques such as Glow-Discharge Optical Emission Spectrometry (GD-OES) and Scanning Electron Microscopy (SEM). Three non-commercial, oil-based lubricants with simplified formulations were used for this study. A tribological test was developed in which two nominally geometrically-identical crossed cylinders slide over each other under selected test conditions. This test was used to evaluate the effectiveness of a pre-applied lubricant film and a surface coating for various conditions of sliding wear. Engineered surface coatings can significantly improve wear resistance of the tool surface but their sliding wear performances strongly depend on the type of coating and lubricant combination used. These coating-lubricant interactions can also have a very strong effect on the useful life of the lubricant in a tribological system. Better performance of lubricants during the sliding wear testing was achieved hen used with the nitrocarburised (CN) coating. To understand the nature of the interactions and their possible effects on the coating-lubricant system, several surface analysis techniques were used. The molecular level investigation of Fourier Transform Infrared Spectroscopy (FTIR) revealed that oxidative degradation occurred in all used oil-based lubricants during the sliding wear test but the degradation behaviour of oil-based lubricants varied with the coating-lubricant system and the wear conditions. The main differences in the carbonyl oxidation region of the FTIR spectra (1900-1600 cm-1) between different coating-lubricant systems may relate to the effective lifetime of the lubricant during the sliding wear test. Secondary Ion Mass Spectrometry (SIMS) depth profiling shows that the CN coating has the highest lubricant absorbability among the tested tool surfaces. Diffusion of chlorine (C1), hydrogen (H) and oxygen (O) into the surface of subsurface of the tool suggested that strong interactions occurred between lubricant and tool surface during the sliding wear test. The possible effects of the interactions on the performance of whole tribological system are also discussed. The study of Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) indicated that the envelope of hydrocarbons (CmHn) of oil lubricant in the positive TOF-SIMS spectra shifted to lower mass fragment after the sliding wear testing due to the breakage of long-chain hydrocarbons to short-chain ones during the degradation of lubricant. The shift of the mass fragment range of the hydrocarbon (CmHn) envelope caries with the type of both tool surface and lubricant, again confirming that variation in the performance of the tool-lubricant system relates to the changes in surface chemistry due to tribochemical interactions at the tool-lubricant interface under sliding wear conditions. The sliding wear conditions resulted in changes not only in topography of the tool surface due to mechanical interactions, as outlined in Chapter 5, but also in surface chemistry due to tribochemical interactions, as discussed in Chapters 6 and 7.

Investigation of the postcure reaction and surface energy of epoxy resins using time-of-flight secondary ion mass spectrometry and contact-angle measurements

Time-of-flight secondary ion mass spectrometry (ToF-SIMS) was used to investigate correlations between the molecular changes and postcuring reaction on the surface of a diglycidyl ether of bisphenol A and diglycidylether of bisphenol F based epoxy resin cured with two different amine-based hardeners. The aim of this work was to present a proof of concept that ToF-SIMS has the ability to provide information regarding the reaction steps, path, and mechanism for organic reactions in general and for epoxy resin curing and postcuring reactions in particular. Contact-angle measurements were taken for the cured and postcured epoxy resins to correlate changes in the surface energy with the molecular structure of the surface. Principal components analysis (PCA) of the ToFSIMS positive spectra explained the variance in the molecular information, which was related to the resin curing and postcuring reactions with different hardeners and to the surface energy values. The first principal component captured information related to the chemical phenomena of the curing reaction path, branching, and network density based on changes in the relative ion density of the aliphatic hydrocarbon and the C₇H₇O⁺ positive ions. The second principal component captured information related to the difference in the surface energy, which was correlated to the difference in the relative intensity of the C_xH_yN_z⁺ ions of the samples. PCA of the negative spectra provided insight into the extent of consumption of the hardener molecules in the curing and postcuring reactions of both systems based on the relative ion intensity of the nitrogen-containing negative ions and showed molecular correlations with the sample surface energy.

Time-of-flight secondary ion mass spectrometry investigation of epoxy resin curing behavior in real time

Time-of-flight secondary ion mass spectrometry and principal components analysis were used in real time to monitor the progress of curing reactions on the surface of a diglycidyl ether of bisphenol A (DGEBA) and diglycidyl ether of bisphenol F (DGEBF) epoxy resin blend reacted with the diamine hardener isophorone diamine at different time intervals. Molecular ions in the mass spectra that characterized the curing reactions steps, including blocking, coupling, branching, and crosslinking, were identified. The aliphatic hydrocarbon ions were correlated to the curing reaction rate, and this indicated that coupling and branching occurred much faster than the blocking and crosslinking curing reactions steps. The total conversion of the coupling and branching reaction steps were followed on the basis of changes with time in the relative ion intensity of molecular ions assigned to the DGEBA/DGEBF, aliphatic hydrocarbon, epoxide, and aromatic ring structures. Indicative measures of crosslinking density were monitored through the observation of changes in the ratio of the relative intensities of the aliphatic hydrocarbon and hydroxyl molecular ions over time. The curing reaction conversion was established by the observation of the changes in the relative ion intensity of the molecular ions that were related to the DGEBA/ DGEBF molecules.

Domino Heck-Aza-Michael reactions : efficient access to 1-substituted tetrahydro-β-carbolines

A simple and efficient palladium-catalyzed domino reaction for the synthesis of a series of C1-substituted tetrahydro-β-carbolines is described. This domino process involves a Heck reaction at the indole 2-position of a halogenated tryptamine precursor, followed by intramolecular aza-Michael addition.