Industry Input

Our team spoke with various experts in the biofuel field: Scott Fenwick the Technical Director of the National Biodiesel Board, Marathon’s Cincinnati Biorefinery Division, and Beckett, a manufacturer of combustion burners. All of these groups of people expressed concerns about the microbiology in biofuels and if adding our microbe would really destroy the biofilms or would just be adding another strand of microbiology into the tanks. Adding another microbe that would not be doing its job would only add to the problems by damaging the biofuel itself, the tank storing the biodiesel, and clogging the fuel filters. Since cinnamaldehyde compromises the membranes of cells our hope is the control the metabolic pathway of cinnamaldehyde to only produce when in close quarters of the biofilm using a regulatory gene. Cinnamaldehyde molecules permeate the membrane of our microbe, fungus, and bacteria in the biofuel; not only can we use cinnamaldehyde to eliminate biofilms, we can also use cinnamaldehyde as a kill switch for our own microbe. Killing off the microbe after its use will control the population of our microbe to ensure safe use of the product.

In this year’s InterLab, our team transformed E. coli DH5α with different plasmids expressing Green Fluorescent Protein (GFP). A plate reader was then used to measure fluorescence and absorbance values from the transformed strains.

Arabinose inducible gp2 (pBAD+gp2)

Along with using cinnamaldehyde to control the population of our microbe, we also researched methods of controlling the population of our microbe using iGEM parts. BBa_K1893016 is a part that inhibits the growth of cell and allows someone to control the population of bacteria. The part is inducible with arabinose and can be easily turned off with the addition of glucose. The part once turned on produces the T7 phage that infects the Escherichia coli causing the lysis of the cell. Parts of this kill switch could be implemented into our project, because controlling the population is very important to not having the microbe create harmful effects in the tank.

AFRL (Dr. Wendy Goodson)

We also considered information conveyed by Dr. Wendy Goodson from the Air Force Research Laboratory in the design of our project. She not only informed us about the composition of biofuels, which steered us to target both fungi and bacteria, but also provided us with fungal and bacterial isolates from fuel tanks. We used the fungal isolates in the testing of our destroy mechanism, and used the knowledge of what bacteria are present in the biofilms to design our detect mechanism.