Assessing the growth kinetics and stoichiometry of Escherichia coli at the single‐cell level
Abstract Microfluidic cultivation and single‐cell analysis are inherent parts of modern microbial biotechnology and microbiology. However, implementing biochemical engineering principles based on the kinetics and stoichiometry of growth in microscopic spaces remained unattained. We here present a no...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley-VCH
2023-01-01
|
| Series: | Engineering in Life Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/elsc.202100157 |
| _version_ | 1828071653674844160 |
|---|---|
| author | Katharina Smaluch Bastian Wollenhaupt Heiko Steinhoff Dietrich Kohlheyer Alexander Grünberger Christian Dusny |
| author_facet | Katharina Smaluch Bastian Wollenhaupt Heiko Steinhoff Dietrich Kohlheyer Alexander Grünberger Christian Dusny |
| author_sort | Katharina Smaluch |
| collection | DOAJ |
| description | Abstract Microfluidic cultivation and single‐cell analysis are inherent parts of modern microbial biotechnology and microbiology. However, implementing biochemical engineering principles based on the kinetics and stoichiometry of growth in microscopic spaces remained unattained. We here present a novel integrated framework that utilizes distinct microfluidic cultivation technologies and single‐cell analytics to make the fundamental math of process‐oriented biochemical engineering applicable at the single‐cell level. A combination of non‐invasive optical cell mass determination with sub‐pg sensitivity, microfluidic perfusion cultivations for establishing physiological steady‐states, and picoliter batch reactors, enabled the quantification of all physiological parameters relevant to approximate a material balance in microfluidic reaction environments. We determined state variables (biomass concentration based on single‐cell dry weight and mass density), biomass synthesis rates, and substrate affinities of cells grown in microfluidic environments. Based on this data, we mathematically derived the specific kinetics of substrate uptake and growth stoichiometry in glucose‐grown Escherichia coli with single‐cell resolution. This framework may initiate microscale material balancing beyond the averaged values obtained from populations as a basis for integrating heterogeneous kinetic and stoichiometric single‐cell data into generalized bioprocess models and descriptions. |
| first_indexed | 2024-04-11T00:56:11Z |
| format | Article |
| id | doaj.art-1797376f4c864345aa4cf19e954faa62 |
| institution | Directory Open Access Journal |
| issn | 1618-0240 1618-2863 |
| language | English |
| last_indexed | 2024-04-11T00:56:11Z |
| publishDate | 2023-01-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | Engineering in Life Sciences |
| spelling | doaj.art-1797376f4c864345aa4cf19e954faa622023-01-05T02:40:22ZengWiley-VCHEngineering in Life Sciences1618-02401618-28632023-01-01231n/an/a10.1002/elsc.202100157Assessing the growth kinetics and stoichiometry of Escherichia coli at the single‐cell levelKatharina Smaluch0Bastian Wollenhaupt1Heiko Steinhoff2Dietrich Kohlheyer3Alexander Grünberger4Christian Dusny5Department of Solar Materials – Microscale Analysis and Engineering Helmholtz‐Centre for Environmental Research – UFZ Leipzig Leizpig GermanyMicroscale Bioengineering IBG‐1: Biotechnology Forschungszentrum Jülich GmbH Jülich GermanyMultiscale Bioengineering Faculty of Technology Bielefeld University Bielefeld GermanyMicroscale Bioengineering IBG‐1: Biotechnology Forschungszentrum Jülich GmbH Jülich GermanyMultiscale Bioengineering Faculty of Technology Bielefeld University Bielefeld GermanyDepartment of Solar Materials – Microscale Analysis and Engineering Helmholtz‐Centre for Environmental Research – UFZ Leipzig Leizpig GermanyAbstract Microfluidic cultivation and single‐cell analysis are inherent parts of modern microbial biotechnology and microbiology. However, implementing biochemical engineering principles based on the kinetics and stoichiometry of growth in microscopic spaces remained unattained. We here present a novel integrated framework that utilizes distinct microfluidic cultivation technologies and single‐cell analytics to make the fundamental math of process‐oriented biochemical engineering applicable at the single‐cell level. A combination of non‐invasive optical cell mass determination with sub‐pg sensitivity, microfluidic perfusion cultivations for establishing physiological steady‐states, and picoliter batch reactors, enabled the quantification of all physiological parameters relevant to approximate a material balance in microfluidic reaction environments. We determined state variables (biomass concentration based on single‐cell dry weight and mass density), biomass synthesis rates, and substrate affinities of cells grown in microfluidic environments. Based on this data, we mathematically derived the specific kinetics of substrate uptake and growth stoichiometry in glucose‐grown Escherichia coli with single‐cell resolution. This framework may initiate microscale material balancing beyond the averaged values obtained from populations as a basis for integrating heterogeneous kinetic and stoichiometric single‐cell data into generalized bioprocess models and descriptions.https://doi.org/10.1002/elsc.202100157Escherichia coligrowthmicrofluidicssingle‐cell analysisspecific growth rateyield coefficient |
| spellingShingle | Katharina Smaluch Bastian Wollenhaupt Heiko Steinhoff Dietrich Kohlheyer Alexander Grünberger Christian Dusny Assessing the growth kinetics and stoichiometry of Escherichia coli at the single‐cell level Engineering in Life Sciences Escherichia coli growth microfluidics single‐cell analysis specific growth rate yield coefficient |
| title | Assessing the growth kinetics and stoichiometry of Escherichia coli at the single‐cell level |
| title_full | Assessing the growth kinetics and stoichiometry of Escherichia coli at the single‐cell level |
| title_fullStr | Assessing the growth kinetics and stoichiometry of Escherichia coli at the single‐cell level |
| title_full_unstemmed | Assessing the growth kinetics and stoichiometry of Escherichia coli at the single‐cell level |
| title_short | Assessing the growth kinetics and stoichiometry of Escherichia coli at the single‐cell level |
| title_sort | assessing the growth kinetics and stoichiometry of escherichia coli at the single cell level |
| topic | Escherichia coli growth microfluidics single‐cell analysis specific growth rate yield coefficient |
| url | https://doi.org/10.1002/elsc.202100157 |
| work_keys_str_mv | AT katharinasmaluch assessingthegrowthkineticsandstoichiometryofescherichiacoliatthesinglecelllevel AT bastianwollenhaupt assessingthegrowthkineticsandstoichiometryofescherichiacoliatthesinglecelllevel AT heikosteinhoff assessingthegrowthkineticsandstoichiometryofescherichiacoliatthesinglecelllevel AT dietrichkohlheyer assessingthegrowthkineticsandstoichiometryofescherichiacoliatthesinglecelllevel AT alexandergrunberger assessingthegrowthkineticsandstoichiometryofescherichiacoliatthesinglecelllevel AT christiandusny assessingthegrowthkineticsandstoichiometryofescherichiacoliatthesinglecelllevel |