Team:SydneyUni Australia

Brief Project Description

Background

• 1,2-dichloroethane (DCA) is part of a family of chlorinated hydrocarbons that are primarily derived from industrial solvents such as tetrachloroethene (PCE) and trichloroethene (TCE). Some of these organochlorines are toxic, carcinogenic and generally nasty.
• DCA is a soluble and mobile contaminant of the groundwater in Botany Bay, Sydney, but also elsewhere around the world.
• Conventional treatment involves significant costs, such as the pumping and heat-stripping of groundwater at Botany Bay. A biological alternative may be cheaper and more effective.
• A suite of techniques (including sampling of contaminated sites, growth in bioreactors with selective conditions and protein engineering) have led to an understanding that there are two primary pathways of DCA-degradation (Fig. 1).

Goals

• Construct a BioBrick-compatible vector inspired by the broad host range vector, pBBR1MCS2.
• Construct and compare two of the proposed pathways of DCA biodegradation.
• Characterise the components of the DCA-degradation pathway for admission into the Registry of Standard Parts.
• (If we get time!) Demonstrate the integration of our the pathway in the chromosome of Pseudomonas stutzeri via natural transformation and site-specific recombination.