|Li, Xin Liang|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: March 24, 2007
Publication Date: March 24, 2007
Citation: Hughes, S.R., Mertens, J.A., Qureshi, N., Cotta, M.A., Bischoff, K.M., Liu, S., Li, X. 2007. Plasmid-based functional proteomic workcell evaluation and characterization of yeast strains with improved growth on xylose expressing Xylanase A(Xyn A) and Celulase F(Cel F) from anaerobic fungi Orpinomyces [abstract]. World Congress on Industrial Biotechnology and Bioprocessing. Seminar 2. p. 19. Technical Abstract: The optimization of various genes is important in cellulosic fuel ethanol production from S. cerevisiae to meet the rapidly expanding need for ethanol derived from hemicellulosic materials. The United States Department of Agriculture, Agricultural Research Service, has developed a fully automated platform for molecular biology routines that accomplishes screening of proteins in 96-well format via in vivo expression for evaluating growth on xylose using xylose isomerase expressing insecticidal peptides for use in crop killing insects, such as corn ear worms. These yeast will be used in cellulosic ethanol production and analyzed for optimum growth on pentose sugars such as xylose. Here we discuss all operations of the automated workcell, including the various expression protocols for profiling and characterization of Xyn A and Cel F optimized open reading frames (ORF) generated by amino acid mutagenesis strategy using the ORF scanning methods used at the USDA. These operations allow rapid production and screening of mutagenized Xyn A and Cel F ORFs with the least amount of plates for optimum expression in yeast using xylose as a feedstock.