The primary objective of this proposal is to explore several interrelated strategies for maximizing overproduction of proteins by genetically modified cells. These strategies are based on a blend of biochemical engineering and molecular biology. The major issues that are addressed include large-scale cell separations, bioreactor development and optimization, protein secretion, and optimal gene expression. The common theme underlying an approach to these issues is that of selective cell separations of bacteria and yeast. The technical research program is divided into four major thrusts: - Fundamentals of selective cell separations. - Continuous fermentations with selective cell recycle. - Optimization of secreted protein production in continuous cultures. - Optimal induction of foreign protein expression. In order to carry out these projects, an interdisciplinary team has been assembled that includes a suspension fluid dynamicist, a bacterial physiologist, a yeast geneticist, a bacterial gene-expression expert, a process control engineer, and a biochemical engineer.
