Open AccessDecoupling Growth and Production by Removing the Origin of Replication from a Bacterial Chromosome
Author(s) -
Marje Kasari,
Villu Kasari,
Mirjam Kärmas,
Arvi Jõers
Publication year - 2022
Publication title -
acs synthetic biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.156
H-Index - 66
ISSN - 2161-5063
DOI - 10.1021/acssynbio.1c00618
Subject(s) - biology , bacteriophage , recombinase , microbiology and biotechnology , lac repressor , exponential growth , genetics , bioprocess , repressor , synthetic biology , escherichia coli , gene , lac operon , gene expression , mathematical analysis , paleontology , mathematics , recombination
Efficient production of biochemicals and proteins in cell factories frequently benefits from a two-stage bioprocess in which growth and production phases are decoupled. Here, we describe a novel growth switch based on the permanent removal of the origin of replication ( oriC ) from the Escherichia coli chromosome. Without oriC , cells cannot initiate a new round of replication, and they stop growing while their metabolism remains active. Our system relies on a serine recombinase from bacteriophage phiC31 whose expression is controlled by the temperature-sensitive cI857 repressor from phage lambda. The reporter protein expression in switched cells continues after cessation of growth, leading to protein levels up to 5 times higher compared to nonswitching cells. Switching induces a unique physiological state that is different from both normal exponential and stationary phases. The switched cells remain in this state even when not growing, retain their protein synthesis capacity, and do not induce proteins associated with the stationary phase. Our switcher technology is potentially useful for a range of products and applicable in many bacterial species for decoupling growth and production.