NSF Award
1410219
In the face of dwindling fossil fuel reserves, a high priority has been placed on the development of strategies capable of producing viable replacements for transportation fuels from renewable resources. Even with a dramatic recent increase in domestic natural gas production in the U.S., renewable fuels remain a priority for countries that lack comparable natural resources, and thus represent a potential area for growth for U.S. biotechnology. The challenges associated with accomplishing this task are extensive, and success in this endeavor ultimately depends upon the ability to generate a good yield of a target molecule in a biological source and an efficient isolation and recovery of the molecule from the source. The focus of this proposal will be twofold: 1) the production of a fuel-like compound in a bacterial host, and 2) the design, screening, and implementation of biocompatible materials to efficiently isolate the compound from the bacteria. As a proposal from a predominantly undergraduate institution, this collaborative approach to an immediate and relevant alternative energy source will help catalyze the development of an undergraduate research program in chemistry at the interface with biology at Dixie State University (DSU), located in the remote southwest corner of Utah. This rural community is comprised of students with diverse backgrounds who, through the proposed research plans, will engage in collaborations with established programs at complementary universities. <br/><br/>With this award, the Chemistry of Life Processes Program in the Chemistry Division and NSF-EPSCoR are funding Drs. Rico Del Sesto (Dixie State University, Utah), Blaine Pfeifer (Univ. Buffalo, NY) and Andrew Koppisch (Northern Arizona Univ.) to address the issues above through multiple approaches. First, new extractants will be synthesized for the purpose of efficient sequestration of biofuel products from biological host systems. The extractants to be produced will be applicable to any host generating relevant isoprenoid compounds, but they will particularly be utilized in this proposal with a heterologous production platform. The known biosynthetic pathway for C32-botryococcene will be transplanted into E. coli. In particular, the proposed E. coli host has been engineered to support high-level production of isoprenoid compounds (reaching g/L levels of the Taxol precursor taxadiene). Furthermore, the new heterologous system will be subjected to a range of readily-available engineering and characterization strategies to optimize production of C32-botryococcene. The overall production process will therefore be complemented by a heterologous production host, and the novel introduction of extraction techniques. In addition to a clear applied goal of high level C32-botryococcene production through E. coli, the proposed plan is dedicated to fundamental knowledge of isoprenoid biosynthesis, extractant synthesis and extraction potential, and the cellular-extractant dynamic that is crucial for the success of this application. As such, the proposal features both science and engineering towards a meaningful and timely problem. The applied and basic themes of the proposal are also mirrored by the collaborative team driving this proposal and their collective expertise available to support and enable the overall production process proposed.
