The isolation, characterisation and development of genetic methods for a new solventogenic strain of Clostridium beijerinckii

• Main Author: Little, Gareth Thomas
• Published: University of Nottingham 2014
• Subjects: 579.3
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.718984

In recent times research has returned to focus on the development of sustainable alternatives to fossil fuel based transport fuels. The drivers for this research are the twin issues of improving supply security and concerns over global warming caused by increased greenhouse gas emissions. For a competing fuel to be economically viable it must be cheap to produce, be compatible with existing fuel infrastructure, and not impact upon conventional agriculture. Butanol produced from the fermentation of waste lignocellulose as a feedstock fits the bill for the requirements of a sustainable fuel production process. Butanol can be used in existing combustion engines with limited modification, while using waste lignocellulose as a feedstock represents a cheap, plentiful, sustainable source of energy. Historically butanol, alongside acetone and ethanol, was produced in an industrial fermentation by the bacterium Clostridium ocetobutylicum during the early 20th Century. The goal of the research presented in this thesis is to isolate a solvent producing strain of Clostridium that can be manipulated for use in an industrial process for the production of butanol. A candidate strain of Clostridium beijerinckii was isolated from garden soil that showed promising levels of solvent production and was able to grow on both hexose and pentose sugars. The genome sequence of the organism was obtained using a combination of two different next-generation sequencing technologies. Tools for the genetic manipulation of the strain including ClosTron disruption, Allele Coupled Exchange, in-frame deletion and random mutagenesis were exemplified. In addition an efficient electrotransformation procedure was developed for this organism that could have wider implications for research on Clostridium spp.