Enhancing the stability of continuous fermentations for platform chemical production

Victoria Outram; Andrew  Yiakoumetti; Charlotte  Green; Rebekah  King; John Ward; Alex  Conradie

doi:10.1016/j.isci.2025.111786

Enhancing the stability of continuous fermentations for platform chemical production

Victoria Outram, Andrew Yiakoumetti, Charlotte Green, Rebekah King, John Ward, Alex Conradie^*

^*Corresponding author for this work

Chemical And Process Engineering

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

6 Downloads (Pure)

Abstract

A significant bottleneck of continuous fermentation is the stability of plasmid-based expression systems. This work tested five plasmid addiction systems based on essential gene complementation in continuous E. coli fermentation. The essential genes tested were infA, ssb, proBA, proC, and dapD and evaluated under phosphate-limited continuous fermentation at two dilution rates (0.033 h−1 and 0.1 h−1) and two temperatures (30°C and 37°C). Of these, plasmids stabilized by infA, ssb, and dapD complementation were segregationally stable under all operating conditions. While a lower dilution rate decreased structural stability, this could be remedied by lowering the temperature. At 0.033 h−1 and 30°C, addiction systems based on proC, dapD, and infA complementation conferred segregational stability with no detriment to structural stability, enabling higher yields at lower dilution rates. This work expands the potential of continuous fermentations for bio-based platform chemical production using plasmid addiction systems to ensure plasmid stability.

Original language	English
Article number	111786
Number of pages	21
Journal	iScience
Volume	28
Issue number	3
Early online date	29 Jan 2025
DOIs	https://doi.org/10.1016/j.isci.2025.111786
Publication status	Published - 21 Mar 2025

Funding

This work was supported by Industrial Biotechnology Catalyst project, ConBioChem: Continuous bio-production of commodity chemicals, funded by Innovate UK, BBSRC and EPSRC (grant BB/N023773/1).

Keywords

continuous fermentation
plasmid addiction system
structural stability
temperature
chemical production

Access to Document

10.1016/j.isci.2025.111786Licence: CC BY 4.0

Outram-etal-IScience-2025-Enhancing-the-stability-of-continuous-fermentations-for-platformFinal published version, 3.3 MBLicence: CC BY 4.0

Enhancing the stability of continuous fermentations for platform chemical production. Plasmid maps and fermentation data
Outram, V. (Creator), Yiakoumetti, A. (Creator), Green, C. (Creator), King, R. (Creator), Ward, J. (Creator) & Conradie, A. (Creator), figshare, 5 Feb 2025
DOI: 10.6084/m9.figshare.23976987
Dataset

Cite this

@article{07326a799fdb486297c0ebb91104b36f,

title = "Enhancing the stability of continuous fermentations for platform chemical production",

abstract = "A significant bottleneck of continuous fermentation is the stability of plasmid-based expression systems. This work tested five plasmid addiction systems based on essential gene complementation in continuous E. coli fermentation. The essential genes tested were infA, ssb, proBA, proC, and dapD and evaluated under phosphate-limited continuous fermentation at two dilution rates (0.033 h−1 and 0.1 h−1) and two temperatures (30°C and 37°C). Of these, plasmids stabilized by infA, ssb, and dapD complementation were segregationally stable under all operating conditions. While a lower dilution rate decreased structural stability, this could be remedied by lowering the temperature. At 0.033 h−1 and 30°C, addiction systems based on proC, dapD, and infA complementation conferred segregational stability with no detriment to structural stability, enabling higher yields at lower dilution rates. This work expands the potential of continuous fermentations for bio-based platform chemical production using plasmid addiction systems to ensure plasmid stability.",

keywords = "continuous fermentation, plasmid addiction system, structural stability, temperature, chemical production",

author = "Victoria Outram and Andrew Yiakoumetti and Charlotte Green and Rebekah King and John Ward and Alex Conradie",

year = "2025",

month = mar,

day = "21",

doi = "10.1016/j.isci.2025.111786",

language = "English",

volume = "28",

journal = "iScience",

issn = "2589-0042",

publisher = "Cell Press",

number = "3",

}

TY - JOUR

T1 - Enhancing the stability of continuous fermentations for platform chemical production

AU - Outram, Victoria

AU - Yiakoumetti, Andrew

AU - Green, Charlotte

AU - King, Rebekah

AU - Ward, John

AU - Conradie, Alex

PY - 2025/3/21

Y1 - 2025/3/21

N2 - A significant bottleneck of continuous fermentation is the stability of plasmid-based expression systems. This work tested five plasmid addiction systems based on essential gene complementation in continuous E. coli fermentation. The essential genes tested were infA, ssb, proBA, proC, and dapD and evaluated under phosphate-limited continuous fermentation at two dilution rates (0.033 h−1 and 0.1 h−1) and two temperatures (30°C and 37°C). Of these, plasmids stabilized by infA, ssb, and dapD complementation were segregationally stable under all operating conditions. While a lower dilution rate decreased structural stability, this could be remedied by lowering the temperature. At 0.033 h−1 and 30°C, addiction systems based on proC, dapD, and infA complementation conferred segregational stability with no detriment to structural stability, enabling higher yields at lower dilution rates. This work expands the potential of continuous fermentations for bio-based platform chemical production using plasmid addiction systems to ensure plasmid stability.

AB - A significant bottleneck of continuous fermentation is the stability of plasmid-based expression systems. This work tested five plasmid addiction systems based on essential gene complementation in continuous E. coli fermentation. The essential genes tested were infA, ssb, proBA, proC, and dapD and evaluated under phosphate-limited continuous fermentation at two dilution rates (0.033 h−1 and 0.1 h−1) and two temperatures (30°C and 37°C). Of these, plasmids stabilized by infA, ssb, and dapD complementation were segregationally stable under all operating conditions. While a lower dilution rate decreased structural stability, this could be remedied by lowering the temperature. At 0.033 h−1 and 30°C, addiction systems based on proC, dapD, and infA complementation conferred segregational stability with no detriment to structural stability, enabling higher yields at lower dilution rates. This work expands the potential of continuous fermentations for bio-based platform chemical production using plasmid addiction systems to ensure plasmid stability.

KW - continuous fermentation

KW - plasmid addiction system

KW - structural stability

KW - temperature

KW - chemical production

U2 - 10.1016/j.isci.2025.111786

DO - 10.1016/j.isci.2025.111786

M3 - Article

SN - 2589-0042

VL - 28

JO - iScience

JF - iScience

IS - 3

M1 - 111786

ER -

Enhancing the stability of continuous fermentations for platform chemical production

Abstract

Funding

Keywords

Access to Document

Fingerprint

Datasets

Enhancing the stability of continuous fermentations for platform chemical production. Plasmid maps and fermentation data

Cite this