| To meet increasing demand for energy from agricultural materials to replace petroleum-based energy sources, fuel ethanol production from agricultural feedstocks must be optimized. USDA-ARS has developed the first fully-automated platform employing molecular biology routines to optimize genes for engineering an industrial Saccharomyces cerevisiae strain to ferment xylose from corn fiber as well as glucose from corn starch. The automated platform is also used to incorporate genes into this yeast to express a commercially-useful, biobased product with pesticidal activity against the corn earworm in addition to expressing xylose isomerase and other genes for fermentation of xylose from hydrolyzed cellulosic biomass. Genes are optimized using an amino acid scanning mutagenesis strategy; the improved yeast strain is then screened for optimal growth on pentose sugars. All operations of the automated workcell will be discussed, including expression protocols for profiling and characterizing the pesticide and the optimized open reading frames. |