PartA: tolerance maximisation

The aim of this part of the project is to test the toxicity of DCM on various species of bacteria. This will allow us to identify strains with a high tolerance to DCM that can then be used in the bioremediation kit. Once we have identified tolerant strains, we will aim to evolve this tolerance further by exposing the bacteria to increasing concentrations of DCM and isolating populations that show an advanced tolerance to it. By using strains with higher tolerance to DCM we will be able to make our kit more robust and efficient. Furthermore, from previous studies it has been conjectured that it is not DCM itself that is toxic to DM4 (and other bacteria) at concentrations >20 mM but an intermediate in the metabolism of DCM. We aim to prove that it is indeed a metabolite in the catabolism of DCM that accounts for the observed toxicity of DCM and identify the metabolite responsible.

This work will also supplement the experiments carried out by other groups within the team. By identifying the metabolite responsible for observed DCM toxicity, we will allow the microcompartment group (Part C) to target their work to one step in metabolism. We will also allow Part A to carry out their work in various strains of bacteria rather than using one chassis. We hope that this will allow Part A to test which strain shows the highest expression and enzymatic activity of dcmA which will allow us to select the most efficient strain and produce the more robust DCM bioremediation system