Microbial stem cell technology
View Patent ↗The present disclosure relates to microbial stem cell technology that enables a growing microbial culture to stably maintain two or more distinct cell types in a ratio that can be genetically programmed and/or dynamically controlled during cultivation. It is contemplated that embodiments described herein can be utilized to increase product yield in microbial fermentations and advanced engineering of biomaterials using genetically engineered microbial cells, among others.
1 . A method of establishing bacterial cell types, comprising:
introducing a nucleic acid sequence into a bacterial cell population,
translationally fusing at least a first signaling protein and a second signaling protein to a PopZ protein, the PopZ protein adapted to asymmetrically localize at a first end of cells within the bacterial cell population, to form a PopZ-signaling factor complex configured to influence cell physiology;
inducing expression of the nucleic acid sequence by observing the first signaling protein;
forming first cells that contain the PopZ-signaling factor complex and second cells that do not contain the PopZ-signaling factor complex via asymmetric cell division of the cells between the first end of the cells and a second end of the cells;
introducing a small molecule in the bacterial cell population to enhance the establishment of the first cells and the second cells by observing the second signaling protein; and
eliciting a different behavior in the first cells containing the PopZ-signaling factor complex and the second cells without the PopZ-signaling factor complex of the bacterial cell population.
2 . The method of claim 1 , wherein the PopZ protein is adapted to asymmetrically localize at a cell pole at the first end of the cells of the bacterial cell population.
3 . The method of claim 1 , wherein the first signaling protein is a phosphodiesterase.
4 . The method of claim 3 , wherein the phosphodiesterase is a c-di-GMP phosphodiesterase YhjH from E. coli.
5 . The method of claim 4 , wherein translationally fusing further comprises translationally fusing the c-di-GMP phosphodiesterase YhjH and a mCherry protein with a N-terminus of the PopZ protein using 9- and 12-amino acid linker sequences to form a tripartite YhjH-mChy-PopZ biochemical control platform.
6 . The method of claim 5 , wherein eliciting a different behavior in the first cells containing the PopZ-signaling factor complex and the second cells without the PopZ-signaling factor complex of the bacterial cell population further comprises eliciting production of wax ester synthase in the first cells containing the tripartite YhjH-mChy-PopZ biochemical control platform and no production of wax ester synthase in the second cells that do not contain the tripartite YhjH-mChy-PopZ biochemical control platform.