<p>The present Master thesis describes the development and optimization of a fed-batch process for production of recombinant proteins in Escherichia coli BL21(DE3) in a multi-fermentor system. The system consists of six 1-liter fermentors, capable of producing 500-1500 μg/mL with present protocol.</p><p>Response surface methodology (RSM) was used for multivariable optimization regarding cultivation time, pH, temperature and feed rate. Optimal protein expression conditions were found out to be 17.8 h cultivation time, 36.7 ºC, pH 6.8 and a feed rate corresponding to specific growth of 0.23 h-1, on glucose substrate. The aggregation of expressed proteins to inclusion bodies, could not be affected by the various growth conditions employed during cultivations.</p><p>A study was conducted regarding growth conditions effect on phosphogluconoylation of expressed proteins. In ten fed-batch cultivations on glucose, LC/MS analysis showed a gluconoylated fraction with additional 178 Da mass, but no correlation between growth conditions and gluconoylation could be found. In two fed-batch cultivations on glycerol-feed, a lower feed rate resulted in no gluconoylation, while a higher did. An explanation would be that the lower amount of available intra-cellular carbon limits formation of gluconoylation precursors.</p>

The aim of this thesis work is to increase the knowledge of phenomena taking place during the initial stage in a top blown converter. The work has been done in a few steps resulting in four different supplements. Water model experiments have been carried out using particle image velocimetry (PIV) technology. The system investigated was a fundamental top blown converter where an air jet was set to impinge on a water surface. The flow field of the combined blown case, where an air jet was introduced through a bottom nozzle, was also captured by the PIV. The work clearly showed that the flow field caused by an impinging top blown jet alone could not match that of the bottom blown case. The main re-circulation loop (or vortex) was investigated with respect to position and it was found that an increased flow rate pushes the center of the re-circulation loop downwards into the bath. However, for the top-blown case there is a point when the flow rate is too large to cause a distinguishable re-circulation loop since the jet becomes more plunging (i.e. penetrates deep into the bath) than impinging, with large surface agitation and splashing as a result.A numerical model with the same dimensions as the experimental system was then created. Three different turbulence models from the same family were tested: standard-, realizable- and a modified-(slight modification of one of the coefficients in order to produce less spreading of the air jet) k-ε turbulence model. It could be shown that for the family of k-ε turbulence models the difference in penetration depth was small and that the values corresponded well to literature data. However, when it comes to the position of the re-circulation loop it was shown that the realizable k-ε model produced better results when comparing the results to the experimental data produced from the PIV measurements, mentioned earlier.It was then shown how the computational fluid dynamics (CFD) model could be coupled to thermodynamics databases in order to solve for both reactions and transport in the system. Instead of an air-water system, a gas-steel-slag system was created using the knowledge obtained in the previous simulation step described above. Reactions between gas-steel, gas-slag, steel-slag and gas-steel-slag were considered. Extrapolation of data from a few seconds of simulation was used for comparison to experimental data from the literature and showed reasonable agreement. The overall conclusion was that it is possible to make a coupling of the Thermo-Calc databases and a CFD software to make dynamic simulations of metallurgical processes such as a top-blown converter.A parametric study was then undertaken where two different steel grades were tested; one with high initial carbon content (3.85 mass-%) and one with lower carbon content (0.5 mass-%). The initial silicon content was held constant at 0.84 mass-%. Different initial temperatures were tested and also some variation in initial dissolved oxygen content was tried. It was found that the rate of decarburization/desiliconization was influenced by the temperature and carbon concentration in the melt, where a high temperature as well as a high carbon concentration favors decarburization over desiliconization. It was also seen that the region affected by a lower concentration of alloys (or impurities) was quite small close to the axis where the impinging jet hits the bath. Add the oscillating nature of the cavity and it was realized that sampling from this region during an experiment might be quite difficult. / QC 20100720

The present Master thesis describes the development and optimization of a fed-batch process for production of recombinant proteins in Escherichia coli BL21(DE3) in a multi-fermentor system. The system consists of six 1-liter fermentors, capable of producing 500-1500 μg/mL with present protocol. Response surface methodology (RSM) was used for multivariable optimization regarding cultivation time, pH, temperature and feed rate. Optimal protein expression conditions were found out to be 17.8 h cultivation time, 36.7 ºC, pH 6.8 and a feed rate corresponding to specific growth of 0.23 h-1, on glucose substrate. The aggregation of expressed proteins to inclusion bodies, could not be affected by the various growth conditions employed during cultivations. A study was conducted regarding growth conditions effect on phosphogluconoylation of expressed proteins. In ten fed-batch cultivations on glucose, LC/MS analysis showed a gluconoylated fraction with additional 178 Da mass, but no correlation between growth conditions and gluconoylation could be found. In two fed-batch cultivations on glycerol-feed, a lower feed rate resulted in no gluconoylation, while a higher did. An explanation would be that the lower amount of available intra-cellular carbon limits formation of gluconoylation precursors.

<p>This study has focused on determining the heat flows in a production blast furnace hearth. This part of the blast furnace is exposed to high temperatures. In order to increase the campaign length of the lining an improved knowledge of heat flows are necessary. Thus, it has been studied both experimentally and numerically by heat transfer modeling. Measurements of outer surface temperatures in the lower part of a production blast furnace were carried out. In the experimental study, relations were established between lining temperatures and outer surface temperatures. These relations were used as boundary conditions in a mathematical model, in which the temperature profiles in the hearth lining are calculated. The predictions show that the corner between the wall and the bottom is the most sensitive part of the hearth. Furthermore, the predictions show that no studied part of the lining had an inner temperature higher than the critical temperature 1150°C, where the iron melt can be in contact with the lining.</p>

During the steelmaking process samples are taken from the liquid steel, mainly to assess the chemical composition of the steel. Recently, methods for rapid determination of inclusion characteristics (size and composition) have progressed to the level where they can be implemented in process control. Inclusions in steel can have either good or detrimental effects depending on their characteristics (size, number, composition and morphology). Thereby, by determination of the inclusion characteristics during the steelmaking process it is possible to steer the inclusion characteristics in order to increase the quality of the steel. However, in order to successfully implement these methods it is critical that the samples taken from the liquid steel represent the inclusion characteristics in the liquid steel at the sampling moment.   The purpose of this study is to investigate the changes in inclusion characteristics during the liquid steel sampling process. Experimental studies were carried out at steel plants to measure filling velocity and solidification rate in real industrial samples. The sampling conditions for three sample geometries and two slag protection types were determined. Furthermore, the dispersion of the total oxygen content in the samples was evaluated as a function of sample geometry and type of slag protection. In addition, the effects of cooling rate as well as oxygen and sulfur content on the inclusion characteristics were investigated in laboratory and industrial samples. Possibilities to separate primary (existing in the liquid steel at sampling moment) and secondary (formed during cooling and solidification) inclusions depending on size and composition were investigated. Finally, in order to evaluate the homogeneity and representative of the industrial samples the dispersion of inclusion characteristics in different zones and layers of the samples were investigated.   It was concluded that the type of slag protection has a significant effect on the filling velocity and the sampling repeatability. Furthermore, that the thickness of the samples is the main controlling factor for the solidification rate. It was shown that top slag can contaminate the samples. Therefore, the choice of slag protection type is critical to obtain representative samples. It was shown that the cooling rate has a significant effect on the number of secondary precipitated inclusions. However, the number of primary inclusions was almost constant and independent on the cooling rate. In most cases it is possible to roughly separate the secondary and primary oxide inclusions based on the particle size distributions. However, in high-sulfur steels a significant amount of sulfides precipitate heterogeneously during cooling and solidification. This makes separation of secondary and primary inclusions very difficult. Moreover, the secondary sulfides which precipitate heterogeneously significantly change the characteristics (size, composition and morphology) of primary inclusions. The study revealed that both secondary and primary inclusions are heterogeneously dispersed in the industrial samples. In general, the middle zone of the surface layer is recommended for investigation of primary inclusions. / QC 20101112

The main objective of this industrially important work was to gain an increasing understanding of the properties of some industrially important materials such as CMSX-4 nickel base super alloy, 90Ti.6Al.4V alloy, 25Cr:6Ni stainless steel, 0.7% carbon steel, AISI 304 stainless steel-alumina composites, mould powder used in continuous casting of steel as well as coke used in blast furnace with special reference to the thermal diffusivities. The measurements were carried out in a wide temperature range covering solid, liquid, glassy and crystalline states. For CMSX-4 alloy, the thermal conductivities were calculated from the experimental thermal diffusivities. Both the diffusivities and conductivities were found to increase with increasing temperature. Microscopic analysis showed the presence of intermetallic phases γ´ such as Ni3Al below 1253 K. In this region, the mean free path of the electrons and phonons is likely to be limited by scattering against lattice defects. Between 1253 K and solidus temperature, these phases dissolved in the alloy adding to the impurities in the matrix, which, in turn, caused a decrease in the thermal diffusivity. This effect was confirmed by annealing the samples at 1573 K. The thermal diffusivities of the annealed samples measured at 1277, 1403 and 1531 K were found to be lower than the thermal diffusivities of non-annealed samples and the values did not show any noticeable change with time. It could be related to the attainment of equilibrium with the completion of the dissolution of γ´ phase during the annealing process. Liquid CMSX-4 does not show any change of thermal diffusivity with temperature. It may be attributed to the decrease of the mean free path being shorter than characteristic distance between two neighbouring atoms. Same tendency could be observed in the case of 90Ti.6Al.4V alloy. Since the thermal diffusivity increases with increasing temperature below 1225 K and shows slight decrease or constancy at higher temperature. For 25Cr:6Ni stainless steel, the thermal diffusivity is nearly constant up to about 700 K. Beyond that, there is an increase with temperature both during heating as well as cooling cycle. On the other hand, the slope of the curve increases above 950 K, which can be due to the increase of bcc phase in the structure. 0.7% carbon steel shows a decrease in the thermal diffusivity at temperature below Curie point, where the structure contains bcc+ fcc phases. Above this point the thermal diffusivity increases, where the structure contains only fcc phase. The experimental thermal conductivity values of these alloys show good agreement with the calculated values using Mills model. Thermal diffusivity measurements as a function of temperature of sintered AISI 304 stainless steel-alumina composites having various composition, viz, 0.001, 0.01, 0.1, 1, 2, 3, 5, 7, 8 and 10 wt% Al2O3 were carried out in the present work. The thermal diffusivity as well as the thermal conductivity were found to increase with temperature for all composite specimens. The thermal diffusivity/conductivity decreases with increasing weight fraction of alumina in the composites. The experimental results are in good agreement with simple rule of mixture, Eucken equation and developed Ohm´s law model at weight fraction of alumina below 5 wt%. Beyond this, the thermal diffusivity/ conductivity exhibits a high discrepancy probably due to the agglomeration of alumina particles during cold pressing and sintering. On the other hand, thermal diffusivities of industrial mould flux having glassy and crystalline states decrease with increasing temperature at lower temperature and are constant at higher temperature except for one glassy sample. The thermal diffusivity is increased with increasing crystallisation degree of mould flux, which is expected from theoretical considerations. Analogously, the thermal diffusivity measurements of mould flux do not show any significant change with temperature in liquid state. It is likely to be due to the silicate network being largely broken down. In the case of coke, the sample taken from deeper level of the pilot blast furnace is found to have larger thermal diffusivity. This can be correlated to the average crystallite size along the structural c-axis, Lc, which is indicative of the higher degree of graphitisation. This was also confirmed by XRD measurements of the different coke samples. The degree of graphitisation was found to increase with increasing temperature. Further, XRD and heat capacity measurements of coke samples taken from different levels in the shaft of the pilot blast furnace show that the graphitisation of coke was instantaneous between 973 and 1473 K. / QC 20100629

Today, there is a high demand on clean steel for high performance materialproperties. Thus, steel producers try to deliver a steel product with the highestquality and cleanliness to the market. The number of parameters that affect thesteel cleanliness may vary depending on the required material properties of thefinal product. However, the non-metallic inclusion characteristics represent one ofthe most important parameters. More specifically, the composition, size, numberand morphology affect steel cleanliness. In this work, selected parameters affectingthe inclusion characteristics were studied using the following methods: i)thermodynamic calculations (including computational thermodynamiccalculations), ii) inclusion determinations using a cross sectional (CS) method (2Dinvestigations) and iii) inclusion determinations using an electrolytic extraction(EE) method (3D investigations). The computational thermodynamic calculations of the slag-steel and inclusion-steelequilibriums were carried out using the Thermo-Calc software. With the help ofthese calculations, the influence of the slag carryover on the top slag, aluminumcontent in steel and sulfur distribution ratio as well as predictions of stable phasesof inclusions were studied. In addition, inclusion determinations of tool steel gradesamples collected during various stages of the ladle treatment in a scrap-based steelplant were carried out using both 2D and 3D methods. Furthermore, inclusiondeterminations of bearing steel grade samples from a runner system after ingotcasting were performed using a 2D metallographic method (CS-method). Also, theINCAFeature software was used, when using cross sectional method, in order tocollect more statistics of the inclusion characteristics. It was found that slag carryover has a large influence on the composition of theactual top slag as well as the aluminum content in the steel as well as the sulfurdistribution ratio. In addition, steel and slag were found to be in “near”-equilibriumconditions, after the completion of the vacuum degassing operation. Furthermore,the composition of small-size inclusions in samples taken from tool steel was foundto be very scattered. Moreover, the composition of the large-size inclusions wasfound to be less scattered. Furthermore, closer to the top slag composition insamples collected after vacuum degassing. Finally, the accuracy of the inclusioncomposition determinations of tool steel samples using the electrolytic extractionmethod was found to be better than for the cross sectional method. The worseaccuracy of the CS-method is due to a considerable effect of matrix elements oninclusion composition. / QC 20100709

In order to meet the need of metallurgical industry in the world, a new MgAlON-BN composite which can be used for example in special refractory nozzles, tubes and break rings for the continuous casting of steel was studied in the present thesis. The aim was to understand the mechanism of synthesis and their physicochemical properties during the application. Thus, the thermodynamic properties, synthesis process, mechanical properties, thermal shock behaviour, thermal diffusivity/conductivity as well as corrosion resistance to molten iron containing oxygen and molten slag of MgAlON and MgAlON-BN composites have been investigated. The Gibbs energy of formation of MgAlON was estimated using the method proposed by Kaufman. The phase stability diagram of Mg-Al-O-N-B was investigated, and consequently the synthesis parameters were determined. MgAlON and MgAlON-BN composites were fabricated by hot-pressing method. The composites obtained this way were characterized by XRD, SEM, TEM and HREM analyses. A Matrix-flushing method was employed in the quantitative XRD analysis for the multi-component samples to understand the mechanism of synthesis. The relationship between mechanical properties and microstructure of the composites was investigated. The experimental results indicated that BN addition has significant influence on the mechanical properties of the composites. These can be explained by the fact that BN has low Young’s modulus, density and non-reactive nature as well as considerable anisotropy of many properties such as thermal expansion, thermal diffusivity/conductivity. Thus, the addition of BN in MgAlON is likely to lead to the presence of microcracks caused by the mismatch of thermal expansion coefficient. The microcracks result in the enhancement of the strength at elevated temperature and thermal shock durability of the composites. Effective thermal conductivities were evaluated from the present experimental results of thermal diffusivities, heat capacity and density. A model suitable for present composites has been derived based on Luo’s model. The predicted lines calculated by the model were in good agreement with experimental results. The reactions between the composites and molten iron as well as the slag were investigated by ‘‘finger’’ experiments and sessile drop experiments. Both experimental results indicated that the BN addition has positive influence on the corrosion resistance. These are attributed to the excellent corrosion resistance of BN to molten iron and slag, such as the higher contact angle between BN substrate and liquid iron and molten slag compared with that obtained for pure MgAlON. / QC 20100929

An experimental investigation of the binary system Co-Cr and the ternary system C-Co-Cr has been carried out in the present thesis. The experimental strategy adopted for the binary system was to measure the thermodynamic activities of Cr, the molar heat capacity as function of time, the phase transformation temperatures with corresponding enthalpies, the Curie transition temperature as well as melting temperatures with corresponding enthalpies. In the ternary system the strategy was to determine the solubility of Co in the Cr7C3 phase as well as the C and Cr contents in the Co rich (fcc) binder phase. The experimental results were compared with atomistic simulations of the solubility of Co in the Cr7C3 phase. Solid state galvanic cell measurements were conducted with both ZrO2-7.5 mol % CaO and CaF2 as the solid electrolyte. In view of possible errors in the measurements with ZrO2-7.5 mol % CaO, as a result of electronic contributions to the conduction of the solid electrolyte, new measurements were conducted with CaF2 as the solid electrolyte. The results indicated that the measured EMF values showed trends which were contrary to the thermodynamic behaviour expected from phase diagram considerations. It was concluded that further detailed experimentation was necessary in order to throw more light on the thermodynamic behaviour of the Co-Cr system. Two different series of DSC measurements were conducted, i.e. one in an atmosphere of pure hydrogen and another in pure argon. In the first investigation, conducted in an atmosphere of pure hydrogen in the temperature interval 318-1660 K, evidence was obtained for the existence of a phase transformation around 900 K in the compositional range 20.7-67.1 wt.% Cr. No indications of such a phase transformation had earlier been seen. In the second investigation, conducted in an atmosphere of pure argon in the temperature interval 298-1823 K, special attention was given to alloys in the Co rich corner of the phase diagram, i.e. 0-10 wt.% Cr. This investigation verified earlier findings of a phase transformations around 900 K in the compositional range 20.7-67.4 wt.% Cr. The magnetic transition temperatures for alloys low in Cr content were also obtained. With the use of the DTA technique the melting temperatures with corresponding enthalpy values for alloys in the compositional range 0.9-7.7 wt.% Cr were obtained. The three-phase triangle fcc+Cr7C3+graphite was investigated at 1373 K, 1423 K and 1473 K. The obtained results showed that the solubility of cobalt in the Cr7C3 phase was significantly higher than previously predicted by thermodynamic calculations. / QC 20100930

The objective of this research work was to develop a methodology for experimentally estimating the interfacial properties at slag-metal interfaces. From previous experiments carried out in the division, it was decided to use surface active elements like sulfur or oxygen to trace any motion at the interface. For this purpose the following experimental investigations were carried out. Firstly the density of slag was estimated using the Archimedes Principle and the Sessile Drop technique. The density of the slag would give the molten slag height required for the surface active element to travel before reaching the slag-metal interface. Diffusivity measurements were uniquely designed in order to estimate the sulfur diffusion through slag media. It was for the first time that the chemical diffusivity was estimated from the concentration in the metal phase. Experiments carried out validated the models developed earlier. The density and diffusivity value of sulfur in the slag was used to accurately capture the time for sulfur to reach the slag-metal interface. The oscillations were identified by calculating the contact angle variations and the interfacial velocity was estimated from the change in the surface area of the liquid iron drop. The interfacial tension was estimated from the contact angles and the interfacial dilatational modulus was calculated. Based on cold model experiments using water as well as mercury, an equation of the dependence of the interfacial shear viscosity on the interfacial velocity and interfacial tension was established. This paved way for the estimation of the interfacial shear viscosity at the slag-metal interface. The present study is expected to have a strong impact on refining reactions in pyometallurgical industries where slag/metal interfaces play an important role. From a fundamental view point, this provides a deeper insight into interfacial phenomena and presents an experimental technique to quantify the same. / QC 20101130