Our team is local to the Newcastle area where we have an active shipping and mining industry. This leads to higher levels of environmental contamination from aromatic hydrocarbons such as BTEX. Current methods for soil bioremediation are costly and time-consuming.
Our team values the ability of microorganisms to adapt to challenging environments and changing carbon sources. We decided to combine these two features to investigate novel and conventional gene clusters within bacteria that utilise BTEX compounds as carbon sources. We then looked to engineer these clusters by testing new combinations of genes from different organisms. This was in order to create a gene or cluster that could be transformed into native microorganisms and allow for biodegradation of BTEX compounds.
Proposed application would be to clean up local soil sites and other contaminated sites around the world. This would lead to the development of a quick and cost-effective method for soil bioremediation.
The benE transporter was the first focus of our project. Claudia cloned genes for the transporter into an E. coli chassis. We aimed to optimise the transporter to import benzoate and confirm it's expression in E. coli . We also focused on the benABCD gene cluster which is well characterised for its ability to matabolise benzoate to catechol. BenA, BenB and BenC form a complex, benzoate-1,2-deoxygenase which mediates the first step in the process. BenD, or benzoate-diol-dehydrogenase, then acts to remove the carboxyl group to give catechol.
We worked to complete a biosensor that could detect the presence and degradation of benzoate in real time via red and green fluorescent proteins.
We also worked towards characterisation of a novel benzene degradation pathway by observing broad molecular trends in response to the presence of benzene.