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The continued growth of world population and food production leads to a continued increase in organic waste, responsible for many environmental and health problems when disposed in the wrong way. In Makkah city, huge amount of raw slaughterhouse waste from the Hajj sacrifice is generated and disposed in landfill without any treatment each year during Hajj. The huge number of pilgrims and a slaughterhouse waste generated from sacrificed animal (2.5 million animals in 2014) results in an environmental and health problems from the landfill area. One of the most applicable and effective methods to treat organic waste materials is anaerobic digestion (AD). In AD, bioactivity of four different groups of microorganisms in sequential metabolic steps under oxygen free conditions is responsible for breaking down complex organic waste into the simplest compounds and producing energy in form of biogas (a mixture of methane and carbon dioxide). However, the process can collapse when the organic waste contains high fatty materials (such as slaughterhouse waste) as long chain fatty acids (LCFAs) produce a strong inhibitory effect on some AD microorganisms. In a batch system, adding a small amount of fat to a large volume of anaerobic seeding sludge in order to provide sufficient amount of anaerobic inoculum could work effectively. However, this will lead to the disadvantage of wasting most of the reactor volume and therefore reducing the capacity to receive fresh organic waste. This research looks into the possibility of optimising the digestion process in order to achieve a good anaerobic digestion process for high fat concentration substrate with a minimum amount of seeding sludge in the reactor. Different strategies were studied to achieve this goal. These included the use of different fat concentrations in co-digestion with vegetable waste, different I/S (inoculum to substrate) ratios vs/vs, different operating conditions of single stage reactors (mesophilic CSTR, thermophilic CSTR, and mesophilic up-flow), and study of different multi stage reactor sets (two, three, and four stages) in order to optimise the digestion process.

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In this research a new rheometer is developed and the principles of the operation are studied in order to find methodologies that enable making measurements of rheological properties of complex fluids in a "one step" manner. The flow field within the T-junction geometry of the device for fluids that exhibit elongational rheometry is non-homogeneous due to the stagnation point. In contrast with the operation of conventional rheometers, data processing from the prototype rheometer involves modelling of the flow field and the construction of the mapping algorithm to connect data obtained from the device with the underlying elastic constitutive parameters of the fluid using inverse approach.

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Although various drugs have been developed to treat different diseases such as cancer, the therapeutic effects of many drugs have been limited by their undesirable properties such as poor solubility, poor bioactivity, rapid clearance in blood and non-specific distribution. Nanoparticles as carriers have received more and more attention in the last two decades due to their ability of overcoming these obstacles and enhancing the therapeutic efficiency of the conventional drugs. In this thesis, various kinds of nanoparticles were developed aiming at improving the therapeutic efficiency and targeted delivery of anti-cancer drug and gene. Curcumin is a promising anti-cancer drug but its applications in cancer therapy are limited due to its poor solubility, short half-life and low bioavailability. In this thesis, magnetic-polymer core-shell nanoparticles based on non-toxic, biocompatible and biodegradable polymers such as silk fibroin, alginate and chitosan were prepared and optimized to improve the uptake efficiency and cell growth inhibition effect of curcumin towards cancer cells. The size, zeta potential, surface morphology, drug loading / release profile, in vitro uptake and growth inhibition effect to cancer and normal cells of these curcumin loaded nanoparticles were investigated. The results indicated that the curcumin loaded particles exhibited enhanced uptake efficiency and growth inhibition effect on MDA-MB-231 cancer cells compared with free curcumin. Higher uptake efficiency and cytotoxicity to MDA-MB-231 cells than normal human dermal fibroblast cells were observed, suggesting they have specific effects against cancer cells. Moreover, in vitro targeted delivery of curcumin to specific areas of cells was achieved with the presence of an external magnetic field, suggesting these magnetic nanoparticles are promising for targeted delivery of drugs to desired sites applying magnetic forces. Apart from drug delivery the applications of magnetic nanoparticles in gene delivery was also investigated. Polyethyleneimine is one of the most efficient non-viral transfection agents for gene delivery due to its high cationic charge density. In this thesis, silk fibroin was selected to fabricate magnetic-silk / polyethyleneimine core-shell nanoparticles and silk-polyethyleneimine nanoparticles for the transfection of an anticancer gene (c-myc antisense oligodeoxynucleotides) into MDA-MB-231 breast cancer cells and human dermal fibroblast cells. The results illustrated that the cytotoxicity of magnetic-silk / polyethyleneimine core-shell nanoparticles was significantly lower than polyethyleneimine coated magnetic nanoparticles which is widely studied as a gene delivery carrier. The magnetic-silk / polyethyleneimine core-shell nanoparticles were capable of delivering c-myc antisense oligodeoxynucleotides into MDA-MB-231 cells and significantly inhibiting the cell growth. Employing magnetic-silk / polyethyleneimine core-shell nanoparticles, high uptake efficiency of c-myc antisense oligodeoxynucleotides was achieved within 20 min via magnetofection. In addition, magnetic-silk / polyethyleneimine core-shell nanoparticles exhibited higher cytotoxic effect against MDA-MB-231 breast cancer cells than normal human dermal fibroblast. Moreover, in vitro targeted delivery of oligodeoxynucleotides can be achieved using magnetic-silk / polyethyleneimine core-shell nanoparticles under a magnetic field.

Ralstonia eutropha H16 (also known as Cupriavidus necator H16) is a non-pathogenic chemolithoautotrophic soil bacterium. It has increasingly gained biotechnological interest for its use as a microbial cell factory for the production of several valuable bio-based chemicals. However the absence of a large repertoire of molecular tools to engineer this organism remains a critical limiting factor to exploiting its full biotechnological potential. Also, adopting established molecular tools applicable to the more notable microbial hosts such as E. coli and Saccharomyces cerevesiae is severely hampered by chassis-incompatibility and functional variability of essential biological parts. The work detailed in this thesis focuses on the development of key molecular tools crucial to improving the biosynthesis of malonyl-CoA - a precursor metabolite required for the biosynthesis of fatty acids and potentially several valuable bio-products in Ralstonia eutropha H16. All molecular tools developed were based on the broad host range (BHR) plasmid vector backbone of pBBR1MCS1 - a R. eutropha H16-compatible vector. Firstly, to facilitate heterologous pathway optimization, a combination of pre-existing and novel methods of genetic modifications were applied to engineer a collection of 42 promoters. Promoter strengths were characterized using a fluorescence-based assay and benchmarked to the dose-dependent activity of an L-arabinose-inducible PBAD promoter. Next, to detect intracellular accumulation of malonyl-CoA, transcriptional factor-based malonyl-CoA-sensing genetic circuits were developed via careful selection from the promoter collection. Thirdly, BHR L-arabinose-inducible λ-Red plasmid vectors were developed for mediating λ-Red-based genome editing. These were first tested in E. coli BW25113 to confirm their functionality and then subsequently tested in R. eutropha H16. Overall, the collection of engineered promoters yielded a 137-fold range of promoter activity and the malonyl-CoA biosensors responded to changing malonyl-CoA concentrations. The BHR λ-Red plasmids showed high recombination efficiency in E. coli BW25113. The molecular tools developed from this work will further facilitate rapid control and regulation of gene expression in R. eutropha, particularly for malonyl-CoA engineering.