Creating a synthetic biology toolbox for Clostridium sp as hosts for bio-butanol and commodity chemical production

The success of industrial biotechnology relies on taking organisms that are already being used in commercial processes and making them better. To do this in a rational way, one needs the tools to be able to introduce new genes and alter existing ones in predictable ways. This is the aim of the field of synthetic biology, where genetic elements such as promoters, regulatory binding sites and terminators can be combined in predictable ways to give reliable results when applied to different genes. This project will develop such tools for an important species of Clostridium that is used by Green Biologics to produce bio-butanol and other commodity chemicals & has arisen directly from a current collaboration between the partners where the lack of these tools has become apparent. The student will gain fundamental insight into gene structure & regulation in this genus, by progressively building an increasingly complete set of characterised ‘parts’. This will start with assessing & altering promoter strength using rational and empirical approaches. This promoter library will then be improved to enable them to be transcriptionally regulated by small molecules via transcription factors. The project will also identify transcription terminators as additional ‘parts’ to be engineered into the clostridial synthetic biology vector system. Testing of these elements will be in the context of our current work in developing ‘plug-&-play’ genetic cassettes that confer phenotypes related to improved feedstock use. The student will spend at least 3 months during the project with the strain development group at Green Biologics, who are based near Oxford (www.greenbiologics.com). This will give the student valuable experience of working in an industrial environment.