Team:Nevada

     When the group first received these results, they were ecstatic, but they knew that there was still hard work ahead. It was important for the team to show that not only were they producing larger amounts of fatty acids, but also fatty acids that were 12 carbons in length, instead of the usual 16 and 18 carbon chains found in the typical cooking oil. “Medium chain fatty acids, such as C-12 fatty acids, can be used not only as a biofuel for automobiles, but can also be used as a jet fuel” stated Jovanna Casas, a fellow researcher and close colleague. The group is in the process of repeating their work using chromatography to determine the size of the fatty acids and also testing different E. coli strains to determine the optimal strain for producing fatty acids.

     Casey Lear and her enzymology team were responsible for developing a classic alcohol dehydrogenase enzymatic assay, but according to Casey, “Only when high concentrations of ethanol were added to the assay was there any absorbance correlating to ADH activity.” The enzymology group concluded that the ADH assay they developed was not sensitive enough to detect ethanol in the low concentration ranges needed by Jovanna’s E. coli group.

     The E. coli group opted to use a more sensitive alcohol oxidase enzyme instead. The team purchased an EnzyChrom Ethanol Detection Kit (alcohol oxidase) from BioAssay Systems to test ethanol production in their sigma70/PDC/ADH operon transformed into the NEB Iq E.coli cell line. “We grew the cultures with the addition of 2% glucose, since glucose can help improve E. coli growth. With this new assay, we were able to detect 0.02% ethanol”.

Growing Cyanobacteria and E. coli together, can it be done? Scientists may soon have an answer.

     The co-culturing apparatus has just been delivered by Matt Bowden, the team’s lead mechanical engineer. The device will allow for growth of E. coli fed by Synechocystis without cross-contamination between the chambers. Stress testing of the device can now begin, in order to determine how to effectively keep it sterile, prevent all leaks, and ensure the water pump can run for the durations needed to culture cyanobacteria.

      The most important of these tests will be to show that we can prevent all E. coli from entering the cyanobacteria chamber, as contamination will quickly smother the cyanobacteria and prevent photosynthesis. Methods will also need to be developed to sterilize the apparatus, as it is not autoclavable in its entirety and cannot be easily assembled in the sterile hoods available to the team. This may involve running the device while pumping bleach through the system followed by a pumping with sterile water. However, it is unknown if all of the tubing will be resistant to this treatment, and other sterilizing chemicals may need to be used. Finally, we need to be sure that our water pump can run for several days at a time without burning out. If the pump cannot last for the time required for Synechocystis to grow, we will need to redesign the apparatus with a sufficient water pump.