Microbial Contamination Control in Bioprocessing
AssigneeThe Regents of the University of California
InventorsJohn D. Coates, Ouwei Wang, Victor M. Reyes-Umana
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Abstract
Biomanufacturing hygiene control in batch and fed-batch bioreactors is provided using a bioreactor employing a chlorite/Cld system, wherein the bioreactor comprises an engineered cultured cell expressing a recombinant cytoplasmic chlorite dismutase (cCld) sufficient to increase chlorite resistance of the cell, and chlorite sufficient to substantially inhibit growth of one or more contaminating microorganisms in the bioreactor yet not substantially inhibit growth of the cell.
Claims15
A bioreactor employing a chlorite/Cld system for hygiene control, the bioreactor comprising a cultured cell expressing a cytoplasmic chlorite dismutase (cCld) sufficient to increase chlorite resistance of the cell, and chlorite sufficient to substantially inhibit growth of one or more contaminating microorganisms in the bioreactor yet permissive to, and not substantially inhibitory of, growth of the cell, wherein the cCld is engineered from a wild-type to lack a functional N-terminal translocation peptide, which results in an inability of the cell to translocate the cCld from the cell cytoplasm into the periplasm.
A method of culturing a cell, comprising:
- a) culturing a cell recombinantly engineered to express a functional cytoplasmic chlorite dismutase (cCld) in a culture medium under conditions whereby the cCld is expressed in said cell, wherein said culture medium comprises one or more contaminating microorganisms, wherein the cCld is expressed in the cell, wherein the cCld is engineered from a wild-type to lack a functional N-terminal translocation peptide, which results in an inability of the cell to translocate the cCld from the cell cytoplasm into the periplasm;
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- b) treating said culture medium with chlorite in an amount sufficient to kill said one or more contaminating microorganisms without killing said cell, wherein the cCld protects the cell from toxic effects of the chlorite.
Dependencies are inferred from the claim text.