Application of a Genetically Encoded Biosensor for Live Cell Imaging of L-Valine Production in Pyruvate Dehydrogenase Complex-Deficient Corynebacterium glutamicum Strains

Mustafi N, Grünberger A, Mahr R, Helfrich S, Nöh K, Blombach B, Kohlheyer D, Frunzke J (2014)
PLoS one 9(1): e85731.

Download
OA 7.38 MB
Zeitschriftenaufsatz | Veröffentlicht | Englisch
Autor
; ; ; ; ; ; ;
Abstract / Bemerkung
The majority of biotechnologically relevant metabolites do not impart a conspicuous phenotype to the producing cell. Consequently, the analysis of microbial metabolite production is still dominated by bulk techniques, which may obscure significant variation at the single-cell level. In this study, we have applied the recently developed Lrp-biosensor for monitoring of amino acid production in single cells of gradually engineered L-valine producing Corynebacterium glutamicum strains based on the pyruvate dehydrogenase complex-deficient (PDHC) strain C. glutamicum ΔaceE. Online monitoring of the sensor output (eYFP fluorescence) during batch cultivation proved the sensor's suitability for visualizing different production levels. In the following, we conducted live cell imaging studies on C. glutamicum sensor strains using microfluidic chip devices. As expected, the sensor output was higher in microcolonies of high-yield producers in comparison to the basic strain C. glutamicum ΔaceE. Microfluidic cultivation in minimal medium revealed a typical Gaussian distribution of single cell fluorescence during the production phase. Remarkably, low amounts of complex nutrients completely changed the observed phenotypic pattern of all strains, resulting in a phenotypic split of the population. Whereas some cells stopped growing and initiated L-valine production, others continued to grow or showed a delayed transition to production. Depending on the cultivation conditions, a considerable fraction of non-fluorescent cells was observed, suggesting a loss of metabolic activity. These studies demonstrate that genetically encoded biosensors are a valuable tool for monitoring single cell productivity and to study the phenotypic pattern of microbial production strains.
Erscheinungsjahr
Zeitschriftentitel
PLoS one
Band
9
Zeitschriftennummer
1
Artikelnummer
e85731
eISSN
PUB-ID

Zitieren

Mustafi N, Grünberger A, Mahr R, et al. Application of a Genetically Encoded Biosensor for Live Cell Imaging of L-Valine Production in Pyruvate Dehydrogenase Complex-Deficient Corynebacterium glutamicum Strains. PLoS one. 2014;9(1): e85731.
Mustafi, N., Grünberger, A., Mahr, R., Helfrich, S., Nöh, K., Blombach, B., Kohlheyer, D., et al. (2014). Application of a Genetically Encoded Biosensor for Live Cell Imaging of L-Valine Production in Pyruvate Dehydrogenase Complex-Deficient Corynebacterium glutamicum Strains. PLoS one, 9(1), e85731. doi:10.1371/journal.pone.0085731
Mustafi, N., Grünberger, A., Mahr, R., Helfrich, S., Nöh, K., Blombach, B., Kohlheyer, D., and Frunzke, J. (2014). Application of a Genetically Encoded Biosensor for Live Cell Imaging of L-Valine Production in Pyruvate Dehydrogenase Complex-Deficient Corynebacterium glutamicum Strains. PLoS one 9:e85731.
Mustafi, N., et al., 2014. Application of a Genetically Encoded Biosensor for Live Cell Imaging of L-Valine Production in Pyruvate Dehydrogenase Complex-Deficient Corynebacterium glutamicum Strains. PLoS one, 9(1): e85731.
N. Mustafi, et al., “Application of a Genetically Encoded Biosensor for Live Cell Imaging of L-Valine Production in Pyruvate Dehydrogenase Complex-Deficient Corynebacterium glutamicum Strains”, PLoS one, vol. 9, 2014, : e85731.
Mustafi, N., Grünberger, A., Mahr, R., Helfrich, S., Nöh, K., Blombach, B., Kohlheyer, D., Frunzke, J.: Application of a Genetically Encoded Biosensor for Live Cell Imaging of L-Valine Production in Pyruvate Dehydrogenase Complex-Deficient Corynebacterium glutamicum Strains. PLoS one. 9, : e85731 (2014).
Mustafi, Nurije, Grünberger, Alexander, Mahr, Regina, Helfrich, Stefan, Nöh, Katharina, Blombach, Bastian, Kohlheyer, Dietrich, and Frunzke, Julia. “Application of a Genetically Encoded Biosensor for Live Cell Imaging of L-Valine Production in Pyruvate Dehydrogenase Complex-Deficient Corynebacterium glutamicum Strains”. PLoS one 9.1 (2014): e85731.
Alle Dateien verfügbar unter der/den folgenden Lizenz(en):
Copyright Statement:
This Item is protected by copyright and/or related rights. [...]
Volltext(e)
Access Level
OA Open Access
Zuletzt Hochgeladen
2017-07-11T10:04:46Z

31 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Technical bias of microcultivation environments on single-cell physiology.
Dusny C, Grünberger A, Probst C, Wiechert W, Kohlheyer D, Schmid A., Lab Chip 15(8), 2015
PMID: 25710324
Vizardous: interactive analysis of microbial populations with single cell resolution.
Helfrich S, Azzouzi CE, Probst C, Seiffarth J, Grünberger A, Wiechert W, Kohlheyer D, Nöh K., Bioinformatics 31(23), 2015
PMID: 26261223
Single-cell microfluidics: opportunity for bioprocess development.
Grünberger A, Wiechert W, Kohlheyer D., Curr Opin Biotechnol 29(), 2014
PMID: 24642389
Application of metabolic engineering for the biotechnological production of L-valine.
Oldiges M, Eikmanns BJ, Blombach B., Appl Microbiol Biotechnol 98(13), 2014
PMID: 24816722
Light-responsive control of bacterial gene expression: precise triggering of the lac promoter activity using photocaged IPTG.
Binder D, Grünberger A, Loeschcke A, Probst C, Bier C, Pietruszka J, Wiechert W, Kohlheyer D, Jaeger KE, Drepper T., Integr Biol (Camb) 6(8), 2014
PMID: 24894989
Metabolic variability in bioprocessing: implications of microbial phenotypic heterogeneity.
Delvigne F, Zune Q, Lara AR, Al-Soud W, Sørensen SJ., Trends Biotechnol 32(12), 2014
PMID: 25457387

Export

Markieren/ Markierung löschen
Markierte Publikationen

Open Data PUB

Web of Science

Dieser Datensatz im Web of Science®

Quellen

PMID: 24465669
PubMed | Europe PMC

Suchen in

Google Scholar