Journal Article

FZJ-2023-03000

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Experimental k S estimation: A comparison of methods for Corynebacterium glutamicum from lab to microfluidic scale

Steinhoff, H. ; Finger, M. ; Osthege, M.FZJ* ; Golze, C. ; Schito, S.FZJ* ; Noack, S.FZJ* ; Büchs, J. ; Grünberger, A. (Corresponding author)

2023
Wiley New York, NY [u.a.]

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Please use a persistent id in citations: doi:10.1002/bit.28345  doi:10.34734/FZJ-2023-03000

Abstract: Knowledge about the specific affinity of whole cells toward a substrate, commonly referred to as kS, is a crucial parameter for characterizing growth within bioreactors. State-of-the-art methodologies measure either uptake or consumption rates at different initial substrate concentrations. Alternatively, cell dry weight or respiratory data like online oxygen and carbon dioxide transfer rates can be used to estimate kS. In this work, a recently developed substrate-limited microfluidic single-cell cultivation (sl-MSCC) method is applied for the estimation of kS values under defined environmental conditions. This method is benchmarked with two alternative microtiter plate methods, namely high-frequency biomass measurement (HFB) and substrate-limited respiratory activity monitoring (sl-RA). As a model system, the substrate affinity kS of Corynebacterium glutamicum ATCC 13032 regarding glucose was investigated assuming a Monod-type growth response. A kS of <70.7 mg/L (with 95% probability) with HFB, 8.55 ± 1.38 mg/L with sl-RA, and 2.66 ± 0.99 mg/L with sl-MSCC was obtained. Whereas HFB and sl-RA are suitable for a fast initial kS estimation, sl-MSCC allows an affinity estimation by determining tD at concentrations less or equal to the kS value. Thus, sl-MSCC lays the foundation for strain-specific kS estimations under defined environmental conditions with additional insights into cell-to-cell heterogeneity.

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Contributing Institute(s):

1. Biotechnologie (IBG-1)

Research Program(s):

1. 2171 - Biological and environmental resources for sustainable use (POF4-217) (POF4-217)
2. DFG project 427899901 - Filamentöse Co-kulturen: vom Screening zur Bioprozessentwicklung (Co-Pilot) (427899901) (427899901)
3. DFG project 427904493 - Communities of niche-optimized strains (CoNoS) – Ein neues Konzept zur Verbesserung der biotechnologischen Produktion von kleinen Molekülen (427904493) (427904493)

Appears in the scientific report 2023

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Database coverage:
; ; ; BIOSIS Previews ; Biological Abstracts ; Clarivate Analytics Master Journal List ; Current Contents - Agriculture, Biology and Environmental Sciences ; Current Contents - Life Sciences ; DEAL Wiley ; Essential Science Indicators ; IF < 5 ; JCR ; Nationallizenz ; SCOPUS ; Science Citation Index Expanded ; Web of Science Core Collection

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