| The skills required for the optimization and control of protein expression in recombinantly engineered microorganisms is of increasing value to chemical engineers and makes them attractive candidates for positions in the biotechnology industry. Therefore, protocols have been developed for use in the Biochemical Engineering Teaching Laboratory. Procedures for batch fermentations in shake flasks, fed-batch fermentations in the New Brunswick BioFlo IIC, SDS-PAGE, absorbance analysis, and media selection are included.; Novel and robust software has been developed for use in the control of fed-batch bioreactors. The software allows the operator of the equipment to easily develop and implement flexible, knowledge-based control schemes based on the environmental variables monitored by the control unit. A section of the operator's manual and descriptions of the controller components and concepts are presented.; A 185 residue segment of the HA2 domain of hemagglutinin has been expressed in the E. coli host Rosetta pRARE in both batch and fed-batch fermentations. Isotopically labeled amino acids have been selectively incorporated into the expressed protein for the purposes of analysis by rotational-echo double resonance (REDOR). The expression optimization procedure that has been followed utilizing the developed protocols is outlined. |
