The proposed work entails optimizing protoplast fusion and genome-shuffling approaches in the bacterium, E. coli. Additionally, gene chips will be used to identify the genes responsible for antibiotic (aa-hydroxamate) tolerant phenotypes, and small network theory will be employed to understand how connectivity and clustering may affect antibiotic tolerance versus fitness. Improving the understanding of genome evolution and the tradeoffs between the yield from individual metabolic pathways and organism fitness is expected to make a positive impact on the metabolic engineering community. The impact follows from more insights being gleaned on how "global" versus "local" optimization is managed at the molecular level by cellular systems. Such insights, in turn, will allow for improved quantitative prediction of the outcome of different metabolic engineering strategies. Understanding better how genomic evolution can lead to antibiotic resistance and organism fitness should also positively impact work on contending with the medical problem of antibiotic resistant bacteria. The plan to integrate research into education and outreach has two objectives. First, outreach activities will be pursued via Women in Engineering, Multicultural Engineering, and high school honors programs. Secondly, undergraduates will be organized into teams that are charged with developing laboratory demonstration modules. Such teams will integrate prior coursework and learn collaboration skills while producing useful courseware.
The overall objective of this research project was to develop new technologies for efficiently engineering complicated cell behaviors. The specific foucs was on the application of new technologies for recombining genes between bacteria, mapping genes to traits, and analyzing genetic data to improve engineering of relevant traits. The educational objectives of this award were focused on the training of students in scientific and technological areas of national need. Further, an emphasis was placed on the recruiting and training of groups traditionally underrepresented in science and engineering. The specific outcomes of this award are itemized below. 1. Research funded by this grant led to the publication of a number (7) of peer-reviewed manuscripts, appearing in high-level scientific journals such as Metabolic Engineering and Applied and Environmental Microbiology. 2. Research funded by this grant was presented at a large number of national scientific meetings, thus providing an additional avenue for dissemination to the broader community. 3. Research and training paid for by this grant enabled the formation of Opxbio, Inc. This company was founded out of the principal investigators lab in 2007 and is located in Boulder, CO. The company has created more than 70 jobs over the past 6 years and has won a number of awards in recognition of their rapid progress towards commercialization. 4. This award paid in part or full for the training of several PhD students (3 female and 1 male) and 5 undergraduate researchers (3 female). This training was in scientific and technical areas typically underrepresented by females and that are considered of broad national importance (sustainable fuels/chemicals).
