Hot isopropanol quenching procedure for automated microtiter plate scale
13C-labeling
Boiling solvent quenching
Corynebacterium glutamicum
Isotopic labeling
Isotopically transient experiment
Lab automation
Metabolic quenching
Microbioreactor cultivation
Journal
Microbial cell factories
ISSN: 1475-2859
Titre abrégé: Microb Cell Fact
Pays: England
ID NLM: 101139812
Informations de publication
Date de publication:
09 May 2022
09 May 2022
Historique:
received:
11
03
2022
accepted:
26
04
2022
entrez:
8
5
2022
pubmed:
9
5
2022
medline:
11
5
2022
Statut:
epublish
Résumé
Currently, the generation of genetic diversity for microbial cell factories outpaces the screening of strain variants with omics-based phenotyping methods. Especially isotopic labeling experiments, which constitute techniques aimed at elucidating cellular phenotypes and supporting rational strain design by growing microorganisms on substrates enriched with heavy isotopes, suffer from comparably low throughput and the high cost of labeled substrates. We present a miniaturized, parallelized, and automated approach to Our results show that a robotic liquid handler is sufficiently fast to generate informative isotopically transient labeling data. Furthermore, the amount of biomass obtained from a sub-milliliter cultivation in a microbioreactor is adequate for the detection of labeling patterns of free amino acids. Combining the innovations presented in this study, isotopically stationary and instationary automated labeling experiments can be conducted, thus fulfilling the prerequisites for
Sections du résumé
BACKGROUND
BACKGROUND
Currently, the generation of genetic diversity for microbial cell factories outpaces the screening of strain variants with omics-based phenotyping methods. Especially isotopic labeling experiments, which constitute techniques aimed at elucidating cellular phenotypes and supporting rational strain design by growing microorganisms on substrates enriched with heavy isotopes, suffer from comparably low throughput and the high cost of labeled substrates.
RESULTS
RESULTS
We present a miniaturized, parallelized, and automated approach to
CONCLUSIONS
CONCLUSIONS
Our results show that a robotic liquid handler is sufficiently fast to generate informative isotopically transient labeling data. Furthermore, the amount of biomass obtained from a sub-milliliter cultivation in a microbioreactor is adequate for the detection of labeling patterns of free amino acids. Combining the innovations presented in this study, isotopically stationary and instationary automated labeling experiments can be conducted, thus fulfilling the prerequisites for
Identifiants
pubmed: 35527247
doi: 10.1186/s12934-022-01806-4
pii: 10.1186/s12934-022-01806-4
pmc: PMC9082905
doi:
Substances chimiques
Amino Acids
0
Carbon Isotopes
0
2-Propanol
ND2M416302
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
78Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : 427904493
Organisme : Deutsche Forschungsgemeinschaft
ID : 427904493
Informations de copyright
© 2022. The Author(s).
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