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Decoupling Growth and Production by Removing the Origin of Replication from a Bacterial Chromosome

[Image: see text] 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 t...

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Autores principales: Kasari, Marje, Kasari, Villu, Kärmas, Mirjam, Jõers, Arvi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9397407/
https://www.ncbi.nlm.nih.gov/pubmed/35798328
http://dx.doi.org/10.1021/acssynbio.1c00618
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author Kasari, Marje
Kasari, Villu
Kärmas, Mirjam
Jõers, Arvi
author_facet Kasari, Marje
Kasari, Villu
Kärmas, Mirjam
Jõers, Arvi
author_sort Kasari, Marje
collection PubMed
description [Image: see text] 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.
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spelling pubmed-93974072023-07-07 Decoupling Growth and Production by Removing the Origin of Replication from a Bacterial Chromosome Kasari, Marje Kasari, Villu Kärmas, Mirjam Jõers, Arvi ACS Synth Biol [Image: see text] 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. American Chemical Society 2022-07-07 2022-08-19 /pmc/articles/PMC9397407/ /pubmed/35798328 http://dx.doi.org/10.1021/acssynbio.1c00618 Text en © 2022 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Kasari, Marje
Kasari, Villu
Kärmas, Mirjam
Jõers, Arvi
Decoupling Growth and Production by Removing the Origin of Replication from a Bacterial Chromosome
title Decoupling Growth and Production by Removing the Origin of Replication from a Bacterial Chromosome
title_full Decoupling Growth and Production by Removing the Origin of Replication from a Bacterial Chromosome
title_fullStr Decoupling Growth and Production by Removing the Origin of Replication from a Bacterial Chromosome
title_full_unstemmed Decoupling Growth and Production by Removing the Origin of Replication from a Bacterial Chromosome
title_short Decoupling Growth and Production by Removing the Origin of Replication from a Bacterial Chromosome
title_sort decoupling growth and production by removing the origin of replication from a bacterial chromosome
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9397407/
https://www.ncbi.nlm.nih.gov/pubmed/35798328
http://dx.doi.org/10.1021/acssynbio.1c00618
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