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This research investigates the influence of pyrolysis on the properties of several biomasses in the context of heating fuel production for the first time. Laboratory-scale torrefaction, pilot-scale pyrolysis and product development trials have been conducted. Laboratory-scale tests found that torrefaction at 350°C for 30 minutes results in a reduction in volatile content of all investigated biomasses (pine, eucalyptus, hardwood, PKS, almond shell and olive stone) from approximately 80% to 30% indicating suitability for inclusion in smokeless fuels. Meanwhile an increase in gross heating value from 18 MJ/kg up to 28 MJ/kg and a concentration in carbon content indicate a shift in properties towards those similar to medium-ranked coals. Pilot-scale investigations reveal PKS as the preferred biomass for inclusion in domestic heating fuels. Following pyrolysis at 700°C PKS was found to exhibit superior mass and energy yields of 27% and 41%, respectively, when compared to olive stone, almond shell and hardwood. Additionally, thermogravimetric techniques have shown PKS chars to be the least intrinsically reactive with burnout times of 160 minutes following pyrolysis at 700 °C compared to only 15 - 30 minutes for equivalent almond shell, olive stone and hardwood chars. Product development trials indicate that briquetted fuels containing PKS char produced at 500°C possess better physical properties at all bio-char inclusion rates (30 - 100 wt. %). Open fire tests demonstrate that intrinsic char reactivity provides a good indication of combustion behaviour with briquettes containing PKS-derived chars resulting in fires lasting up to 100 minutes longer than those containing almond shell and olive stone chars. Finally, NMR investigations have demonstrated that torrefaction at 350°C results in an increase in aromatic carbon content of biomasses of up to 56% and in-situ NMR techniques have been used for the first time to follow the real-time high-temperature curing process of molasses.

Driven by the diminishing total reserve of fossil fuels and growing concerns about energy security and environmental issues, there is an increasing interest in developing microbial-based processes for the production of biofuel. As most natural fermentation processes do not yield desirable products at an economically viable scale, it is often required to genetically engineer, including deletion and/or insertion of necessary pathways to optimize yield or get new products. Thus it is essential to have the necessary genetic tools, and such was developed, to both improve and innovate upon existing methods, for the process organism of TMO Renewables Ltd - G. thcrmoglucosidasius NCIBM 11955, during this study, with the aim of ultimately creating a thermophilic butanol producing strain. Taking a synthetic biology approach, adopting the clostridial modular shuttle vector format, a new shuttle vector for G. thermoglucosidasius - pMTL86551 was constructed. Based on pMTL86551, along with the development of an appropriate media, a G. thermoglucosidasius pyrE' strain was generated. Utilizing of the pyrE' phenotype both as a positive and negative selection marker allowed successful implementation of rapid procedures for gene knock-in and knock-out by allelic exchange. While the re-creation of the TMO production strain 1 \95SAldhApflpdfp within two month proved the rapidity and feasibility of the new genetic methods, the possibility of developing a thermophilic butanol producing strain, motivated by the superior fuel properties of butanol compared to ethanol, was also opened up. However, implementation of a functional ABE fermentation pathway from a thermophilic clostridium was met with a multitudes of barriers. Moreover, the production strain failed to show the desired fermentation profile. Further research efforts will be required to tackle these problems for progression.

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