Proteomic investigation of elicited Eschscholzia californica cultures producing enhanced levels of benzophenanthridine alkaloids

BIOT 274

John T. Oldham1, Marina Hincapie2, Tomas Rejtar2, Jennifer G. Dy3, and Carolyn WT. Lee-Parsons, clee@coe.neu.edu1. (1) Department of Chemical Engineering, Northeastern University, 360 Huntington Avenue, 342 Snell Engineering Center, Boston, MA 02115, (2) Barnett Institute of Chemical and Biological Analysis, Northeastern University, Boston, MA 02115, (3) Department of Electrical and Computer Engineering, Northeastern University, Boston, MA 02115
Recent developments in proteomics have enabled the global protein profiling of organisms with unsequenced genomes, including many medicinal plants. Eschscholzia californica produces biologically active benzophenanthridine alkaloids (BPAs) and was used as a model system to investigate the complex metabolic mechanisms leading to alkaloid production using proteomics. Cells grown without the hormone 2,4-dichlorophenoxy acetic acid were elicited with purified yeast extract leading to an 18-fold increase in BPAs compared to untreated cultures. Protein extracts of elicited and untreated cultures were separated by HPLC, which revealed differences in protein abundance. HPLC fractions were further separated by gel electrophoresis followed by in-gel digestion of individual bands. Peptide digests were analyzed using MALDI-TOF tandem mass spectrometry and de novo sequencing. Proteins were identified by homology searching against the plant database. Identification of differentially expressed proteins will provide insight for increasing secondary metabolism in plant cell culture.
 

Poster Session
5:30 PM-7:30 PM, Wednesday, August 22, 2007 BCEC -- Exhibit Hall - B2, Poster

Division of Biochemical Technology

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007