Applied and Environmental Microbiology
Thayer School of Engineering
We describe the development of a novel protein expression system based on the industrial fermentation organism Ralstonia eutropha (formerly known as Alcaligenes eutrophus) NCIMB 40124. This new system over- comes some of the shortcomings of traditional Escherichia coli-based protein expression systems, particularly the propensity of such systems to form inclusion bodies during high-level expression. Using a proteomics approach, we identified promoters that can be induced by simple process parameters or medium compositions in high-density cell culture or shake flasks, respectively. By combining newly developed molecular biological tools with a high-cell-density fermentation process, we were able to produce high levels (>1 g/liter) of soluble, active organophosphohydrolase, a model enzyme prone to inclusion body formation in E. coli.
Dartmouth Digital Commons Citation
Srinivasan, Sriram; Barnard, Gavin C.; and Gerngross, Tillman U., "A Novel High-Cell-Density Protein Expression System Based on Ralstonia eutropha" (2002). Dartmouth Scholarship. 497.