Continuous Recombinant Bacterial Fermentations Utilizing Selective Flocculation and Recycle

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Abstract
Selective recycle has successfully been used to maintain an unstable plasmid‐bearing bacterial strain as dominant in a continuous reactor, whereas the culture reverts to 100% segregant cells when selective recycle is not used. The plasmid‐bearing strain is slower growing and flocculent; however, when the cells lose their plasmid, the resulting segregant cells are nonflocculent and grow at a faster rate due to their decreased metabolic burden. Both types of cells exit a chemostat and enter an inclined settler where the flocculent plasmid‐bearing cells are separated from the nonflocculent segregant cells by differential sedimentation. The underflow from the cell separator, which is enriched with plasmid‐bearing cells, is recycled back to the chemostat, while the segregant cells are withdrawn off the top of the setter and discarded. The experimental results agree well with selective recycle reactor theory. On the basis of the theory, a criterion is presented that has been shown to successfully predict whether or not a selective recycle reactor can maintain a plasmid‐bearing strain.