Aqueous-biphasic organometallic catalysis is, as illustrated by the industrial hydroformylation of propene and butene, one of the most promising ways to overcome the intrinsic problem of catalyst separation in organometallic catalysis. However, for poorly water-soluble substrates, mass transfer limitations bring the reaction rate below any that could be economically viable, greatly limiting the scope of this elegant technology. We have studied three different strategies to overcome this limitation. We developed additives that speed up the reaction whilst retaining fast phase separation and good metal retention. Evidence suggests that those additives affect the reaction by forming emulsions with poor stability under the reaction conditions These emulsions increase the interfacial surface area but break after settling for a short time. We also developed ligands that allow the catalyst to be reversibly transported between an aqueous and an organic phase upon addition and removal of carbon dioxide. This allows the reaction to be carried out under homogeneous conditions, only limited by intrinsic kinetics, and the catalyst to be separated by aqueous extraction triggered by carbon dioxide. The catalyst can be returned to a fresh organic phase by flushing out the carbon dioxide. By applying this methodology for the hydroformylation of medium chain length alkenes, very high reaction rates were obtained and the catalyst could be recycle three times with excellent retention of activity and low metal leaching. This methodology could also be reversed with the reaction being carried out in an aqueous phase in the presence of carbon dioxide and extracting the catalyst into an organic solvent using nitrogen flushing. Finally, we briefly investigated the use of an oscillatory baffled reactor as a mean for mass transfer improvement for aqueous-biphasic hydroformylation. This new type reactor did not improve the performance of the system under the investigated conditions, but may require less energy input for equivalent agitation and mixing.

published_or_final_version / Environmental Management / Master / Master of Science in Environmental Management

published_or_final_version / Urban Planning / Master / Master of Science in Urban Planning

No description available.

Bacteria are enclosed by a peptidoglycan sacculus, an exoskeleton-like polymer composed of glycan strands cross-linked by short peptides. The sacculus surrounds the cell in a closed bag-like structure and forms the main structural component of the bacterial cell wall. As bacteria grow and divide, cell wall remodelling by peptidoglycan hydrolases results in the release of peptidoglycan fragments from the sacculus. In Gram-negative bacteria, these fragments are efficiently trapped and recycled. Gram-positive bacteria however shed large quantities of peptidoglycan fragments into the environment. For nearly five decades, Gram-positive bacteria were thus assumed not to recycle peptidoglycan and this process has remained enigmatic until recently. In this thesis, the occurrence and physiological role of peptidoglycan recycling in the Gram-positive pathogen Staphylococcus aureus was investigated. S. aureus is an important pathogen, and is becoming increasingly resistant to many antibiotics. Through bioinformatic and experimental means it was determined that S. aureus may potentially recycle components of peptidoglycan and novel peptidoglycan recycling components were identified and characterised. Though disruption of putative peptidoglycan recycling in S. aureus appears not affect growth or gross morphology of this bacterium, potential roles for peptidoglycan recycling in cell wall homeostasis and in virulence were identified. This is to my knowledge the first demonstration of a potential role of peptidoglycan recycling in either of these aspects of bacterial physiology in any Gram-positive bacterium. This is an important step forward in understanding the basic biology of Gram-positive bacteria, and in understanding the mechanisms of virulence in S. aureus. Future study of this process in S. aureus and other Gram-positive bacteria promises to reveal yet further facets of this process and its functions, potentially leading to the identification of novel therapeutic approaches to combat infections.

In this thesis we investigate advanced techniques for the readout and control of various interferometers. In particular, we present experimental investigations of interferometer configurations and control techniques to be used in second generation interferometric gravitational wave detectors. We also present a new technique, tilt locking, for the readout and control of optical interferometers. ¶ We report the first experimental demonstration of a Sagnac interferometer with resonant sideband extraction (RSE). We measure the frequency response to modulation of the length of the arms and demonstrate an increase in signal bandwidth of by a factor of 6.5 compared to the Sagnac with arm cavities only. We compare Sagnac interferometers based on optical cavities with cavity-based Michelson interferometers and find that the Sagnac configuration has little overall advantage in a cavity-based system. ¶ A system for the control and signal extraction of a power recycled Michelson interferometer with RSE is presented. This control system employs a frontal modulation scheme requiring a phase modulated carrier field and a phase modulated subcarrier field. The system is capable of locking all 5 length degrees of freedom and allows the signal cavity to be detuned over the entire range of possibilities, in principle, whilst maintaining lock. We analytically investigate the modulation/demodulation techniques used to obtain these error signals, presenting an introductory explanation of single sideband modulation/demodulation and double demodulation. ¶ This control system is implemented on a benchtop prototype interferometer. We discuss technical problems associated with production of the input beam modulation components and present several solutions. Operation of the interferometer is demonstrated for a wide range of detunings. The frequency response of the interferometer is measured for various detuned points and we observe good agreement with theoretical predictions. The ability of the control system to maintain lock as the interferometer is detuned is experimentally demonstrated. ¶ Tilt locking, a new technique to obtain an error signal to lock a laser to an optical cavity, is presented. This technique produces an error signal by efficient measurement of the interference between the TEM00 and TEM10 modes. We perform experimental and theoretical comparisons with the widely used Pound-Drever-Hall (PDH) technique. We derive the quantum noise limit to the sensitivity of a measurement of the beam position, and using this result calculate the shot noise limited sensitivity of tilt locking. We show that tilt locking has a quantum efficiency of 80%, compared to 82% for the PDH technique. We present experimental demonstrations of tilt locking in several applications including frequency stabilisation, continuous-wave second harmonic generation, and injection locking of a Nd:YAG slab laser. In each of these cases, we demonstrate that the performance of tilt locking is not the limiting factor of the lock stability, and show that it achieves similar performance to the PDH based system. ¶ Finally, we discuss how tilt locking can be effectively applied to two beam interferometers. We show experimentally how a two beam interferometer typically gives excellent isolation against errors arising from changes in the photodetector position, and experimentally demonstrate the use of tilt locking as a signal readout system for a Sagnac interferometer.

No description available.

The studies reported in this thesis address, firstly, aspects of ribosome recycling in eubacteria, and secondly, a preliminary characterization of an EFG-like locus from Mycobacterium smegmatis. A hitherto unsuspected role of the ribosome recycling factor in governing the fidelity of initiation has been discovered during the course of this work. A summary of the relevant literature is presented in chapter 1. Section I of the ‘General Introduction’ provides a brief review of the current understanding of protein biosynthesis, with a special emphasis on ribosome recycling and the fidelity of translation initiation. Section II provides a brief introduction to mycobacterial translation, and known deviations from the E. coli prototype are highlighted. This is followed by three chapters containing experimental work, as summarized below. (i) Role of elongation factor G in governing specificity of ribosome recycling In eubacteria and the eukaryotic organelles, the post-termination ribosome complexes are recycled by the combined action of ribosome recycling factor (RRF) and elongation factor G (EFG). Earlier studies both from our laboratory and other laboratories have revealed the existence of specific interactions between RRF and EFG that are crucial for ribosome recycling, using ribosomes from E. coli and factors from both E. coli and heterologous sources such as Mycobacterium tuberculosis, Thermus thermophilus etc. In this study, to further understand the mechanism of ribosome recycling, we employed polysomes from both E. coli and M. smegmatis and monitored ribosome recycling in in vitro assays using RRF and EFG from both these sources; in addition, in vivo assays were performed in E. coli using either temperature-sensitive strains or strains carrying a deletion in frr (encoding RRF) or fusA (encoding EFG) genes. It was found that, in E. coli, RRF from Mycobacterium tuberculosis and M. smegmatis function with MtuEFG or MsmEFG but not with EcoEFG. In vitro assays revealed that the mycobacterial EFGs facilitate recycling of both the mycobacterial and E. coli polysomes not only with mycobacterial RRFs but also with EcoRRF. In contrast, although EcoEFG binds to mycobacterial polysomes, carries out GTP hydrolysis and is reported to sustain translocation on mycobacterial ribosomes, its activity in recycling mycobacterial polysomes was undetectable with EcoRRF, as well as with the mycobacterial RRFs. Such an observation allowed us to infer that EFG establishes specific interactions with the ribosome that are crucial for ribosome recycling but not for translocation, suggesting that translocation and ribosome recycling are distinct functions of EFG. In addition, a number of EFG chimeras generated by swapping corresponding domains between Msm- and Eco-EFGs were analyzed for their ability to sustain translocation and/or ribosome recycling in E. coli and M. smegmatis, using a combination of in vivo (for E. coli) and in vitro (for both E. coli and M. smegmatis) approaches. Our observations reveal that a dual set of specific interactions of EFG with RRF and ribosome is essential for ribosome recycling. While the RRF-EFG specific interactions are predominantly localized to the domains IV and V of EFG, the EFG-ribosome specific interactions that are crucial for ribosome recycling are not localized to a specific region of EFG but are found throughout the molecule. Our novel observations also emphasize the importance of using ribosomes from heterologous sources to understand the mechanism of this crucial process. (ii) Impact of rRNA methylations on ribosome recycling and fidelity of initiation in Escherichia coli Ribosomal RNA (rRNA) contains a number of modified nucleosides in functionally important regions including the intersubunit bridge regions; however, very little is known about the role of these rRNA modifications in ribosome function. As the activity of ribosome recycling factor (RRF) in separating the large and the small subunits of the ribosome involves disruption of the intersubunit bridges, we investigated the impact of rRNA methylations on ribosome recycling. The isolation of a folD122 mutant strain of E. coli with a deficiency in rRNA methylations, as well as the availability of E. coli strains deficient for various individual methyltransferases that modify specific rRNA residues, provided us with a genetic tool to assay the role of rRNA methylations in ribosome recycling. We observed that deficiency of rRNA methylations, especially at positions 1518 and 1519 of 16S rRNA near the interface with the 50S subunit and in the vicinity of the IF3 binding site, adversely affects the efficiency of RRF-mediated ribosome recycling. In addition, a compromise in the RRF activity was found to afford increased initiation with a mutant tRNAfMet wherein the three consecutive G-C base pairs (29GGG31:39CCC41), a highly conserved feature of the initiator tRNAs, were mutated to those found in the elongator tRNAMet (29UCA31:39ψGA41). This observation has allowed us to uncover a new role of RRF as a factor that contributes to fidelity of initiator tRNA selection on the ribosome. In addition, it was also found that IF3 and rRNA methylations, both of which are known to affect fidelity of initiation, exert their effects through distinct mechanisms, despite the proximity of a cluster of methylated rRNA residues to the IF3 binding site on the 30S subunit. (iii) Characterization of the role of EFG2, an EFG-like locus in Mycobacterium smegmatis Several bacteria, including various species of mycobacteria (with the exception of Mycobacterium leprae) contain a second EFG-like locus, denoted as fusA2, which shows considerable homology to fusA (encoding EFG). A comparison of the sequences of EFG and EFG2 from various bacteria reveals that EFG2 contains a GTPase domain and domains with significant homology to EFG domains IV and V, suggesting that it may function as an elongation factor. With the single exception of a recent study on Thermus thermophilus EFG2, this class of EFG-like protein factors has not been studied so far. Hence, it was of interest to characterize EFG2. In the current study, EFG2 from M. smegmatis was characterized both by in vitro biochemical assays as well as by in vivo experiments targeted to investigate the biological significance of EFG2 in mycobacteria. It was found that, unlike EFG, MsmEFG2 could not sustain either translocation or ribosome recycling in E. coli. Despite the fact that the purified MsmEFG2 could bind guanine nucleotides, it lacked the ribosome-dependent GTPase activity characteristic of EFG and other translation GTPases, suggesting that it was unlikely to function as an elongation factor. However, EFG2 was found to be expressed in stationary phase cultures of M. smegmatis. To understand the biological significance of EFG2, fusA2 was disrupted in M. smegmatis. The viability of the M. smegmatis mc2155 fusA2::kan derivative indicates that MsmfusA2 is a non-essential gene. While disruption of the fusA2 gene (encoding EFG2) in M. smegmatis does not appear to affect its growth and survival in log phase or stationary phase or under hypoxic conditions, preliminary experiments indicate that disruption of fusA2 confers a fitness disadvantage to M. smegmatis when competed against M. smegmatis mc2155 (with wild type fusA2 locus).

Die eingereichte Dissertation mit dem Thema „Heterogen katalysierte Hydrodehalogenierung von Borhalogeniden im Rahmen eines in sich geschlossenen BNHx-Recyclingkonzepts“ beinhaltet drei für das BNHx-Recycling zentrale Reaktionsschritte: Supersäureaufschluss, Hydrodehalogenierung und den Basenaustausch. Mit Hilfe dieser drei Schritte ist es möglich, die Zersetzungsrückstände von BH3NH3(Polyaminoboran, Borazin und Polyborazylen) wieder in BH3NH3 zu überführen, ohne teure Reduktionsmittel wie LiAlH4 oder Hydrazin verwenden zu müssen. Das Verfahren ermöglichte in einem Durchlauf die Gewinnung von 60 % der eingesetzten Menge an BH3NH3 ohne eine Erzeugung von Abfallprodukten. Mit Hilfe der Chloralkalielelektrolyse kann das Verfahren an Wind- bzw. Solarkraftwerke gekoppelt und über diese alle benötigten Ausgangsstoffe hergestellt werden. Die katalytische Erzeugung von B-H-Spezies auf der Grundlage der Aktivierung von molekularem Wasserstoff ist als Schwerpunkt dieser Arbeit zu sehen. Die mechanistischen Studien lieferten ein genaueres Verständnis über fundamentale Zusammenhänge zwischen dem Lösungsmittel Et3N und der Wasserstoffbereitstellung über die N-CH2-Gruppierung sowie der Wirkungsweise des Katalysators.

There has been considerable interest in finding and explaining the basic elements that can drive product quality up. In the literature this is largely done by modelling the effects of investing in learning and process improvement, and of cost reduction. In the first essay, demand is modelled as a function of price and quality. With this demand function, the firm should produce output of higher quality, the increase in quality being dependent on consumers’ sensitivity to quality and to price, and the effect of technological improvement on product price and quality are very different from those when the demand is a function of price alone. Some twenty states in the U.S. have passed recycling laws which mandate consumption of old newspaper by the newsprint industry. To study the effect of regulation, a model is used in which two firms compete under the regulatory constraint—one firm producing the recycled product, the other the virgin product. Assuming the regulatory constraint is binding, and the demand for the recycled product is derived solely from the legislation, interesting results such as the two firms share equal profits, and consumers pay higher average price in competitive equilibrium than the cartel price, are obtained in the second essay. The two firm model is generalized to include n firms which compete under the same kind of regulatory constraint in the third essay. Results similar to the two firm case are obtained. When the recycled product and the virgin product are partially substitutable, regulation that mandates consumption of the recycled product results in infinitely many equilibria. A dominating equilibrium exists if the demand parameters satisfy a certain condition, otherwise it is not clear how to select an equilibrium. On the other hand, a suitable tax on the virgin product, or its producer, serves to induce compliance with the recycling policy and equilibrium selection. The equilibrium prices and profits of the two firms under the schemes of production tax, excessive consumption tax and progressive profit tax are examined and compared in the fourth essay. It is interesting to find that the tax rate for excessive consumption is comparatively low and, in equilibrium, this tax scheme collects no tax payment.