Team Alberta’s divided approach allowed for simultaneous production of methods to characterize and lay the foundations for a unified project. Our plan was to use cellulose by-products to produce a viable fuel. The questions we sought to answer were:

  1. What organism can we utilize that already consumes cellulose?
  2. How can this organism be modified to increase its fatty acid content?
  3. How can we make genetic modifications easy and accessible for this organism?
  4. How can we use our organism’s outputs to make a viable fuel?
  5. How could our biodiesel affect populations?

The growth section of our project successfully conveyed N. crassa's ability to utilize cellulose by using grass clippings and wheat straw as characterized examples. We found the growth rates of Neurospora on these substrates comparable to that of Neurospora on Vogel's media (VSuTB).

Our project aimed at disrupting beta-oxidation (fatty acid breakdown) and upregulating fatty acid synthesis. We aimed to accomplish this by inserting a thioesterase into the fatty acyl CoA synthetase gene site of Neurospora, thereby simultaneously knocking out an essential gene to beta oxidation and inserting a gene to aide in fatty acid synthesis. We soon learned that constructing genes for Neurospora was not a well-defined or simple process, which is why Team Alberta spent the majority of the summer developing a rapid method to create genes for Neurospora based on our previous experiences in gene construction. These innovations lead to a new assembly method and the creation of BBF RFC 82.

An important aspect to our project was achieving a usable fuel. We developed an efficient and direct method of chemical esterification to effectively allow the fatty acids of Neurospora to be changed into fatty acid methyl esters, an accepted form of biodiesel. We were able to characterize the output of Neurospora fatty acid production as mainly C16 and C18 fatty acids.

Using our data from the wild type Neurospora crassa, we conducted a cost analysis to determine how economical our biodiesel would be and achieved great results for the average savings in cities around the world.

Team Alberta has worked hard over the summer and is proud to be presenting our multifaceted project. We are excited to be introducing Neurospora crassa to the iGEM community and to share our many achievements in the development in this organism.