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.

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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.
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PLoS one
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e85731
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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.
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32 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Small molecule signaling, regulation, and potential applications in cellular therapeutics.
McNerney MP, Styczynski MP., Wiley Interdiscip Rev Syst Biol Med 10(2), 2018
PMID: 28960879
Microbioreactor Systems for Accelerated Bioprocess Development.
Hemmerich J, Noack S, Wiechert W, Oldiges M., Biotechnol J 13(4), 2018
PMID: 29283217
In vivo biosensors: mechanisms, development, and applications.
Shi S, Ang EL, Zhao H., J Ind Microbiol Biotechnol 45(7), 2018
PMID: 29380152
Diverse genetic error modes constrain large-scale bio-based production.
Rugbjerg P, Myling-Petersen N, Porse A, Sarup-Lytzen K, Sommer MOA., Nat Commun 9(1), 2018
PMID: 29463788
Production of L-valine from metabolically engineered Corynebacterium glutamicum.
Wang X, Zhang H, Quinn PJ., Appl Microbiol Biotechnol 102(10), 2018
PMID: 29594358
Coarse-graining bacteria colonies for modelling critical solute distributions in picolitre bioreactors for bacterial studies on single-cell level.
Westerwalbesloh C, Grünberger A, Wiechert W, Kohlheyer D, von Lieres E., Microb Biotechnol 10(4), 2017
PMID: 28371389
Production of amino acids - Genetic and metabolic engineering approaches.
Lee JH, Wendisch VF., Bioresour Technol 245(pt b), 2017
PMID: 28552565
Taking control over microbial populations: Current approaches for exploiting biological noise in bioprocesses.
Delvigne F, Baert J, Sassi H, Fickers P, Grünberger A, Dusny C., Biotechnol J 12(7), 2017
PMID: 28544731
Homogenizing bacterial cell factories: Analysis and engineering of phenotypic heterogeneity.
Binder D, Drepper T, Jaeger KE, Delvigne F, Wiechert W, Kohlheyer D, Grünberger A., Metab Eng 42(), 2017
PMID: 28645641
Maximizing the stability of metabolic engineering-derived whole-cell biocatalysts.
Kadisch M, Willrodt C, Hillen M, Bühler B, Schmid A., Biotechnol J 12(8), 2017
PMID: 28719144
Engineering Microbial Metabolite Dynamics and Heterogeneity.
Schmitz AC, Hartline CJ, Zhang F., Biotechnol J 12(10), 2017
PMID: 28901715
Beyond the bulk: disclosing the life of single microbial cells.
Rosenthal K, Oehling V, Dusny C, Schmid A., FEMS Microbiol Rev 41(6), 2017
PMID: 29029257
Transcription factor-based biosensors in biotechnology: current state and future prospects.
Mahr R, Frunzke J., Appl Microbiol Biotechnol 100(1), 2016
PMID: 26521244
Microfluidics and microbial engineering.
Kou S, Cheng D, Sun F, Hsing IM., Lab Chip 16(3), 2016
PMID: 26758660
Updates on industrial production of amino acids using Corynebacterium glutamicum.
Wendisch VF, Jorge JMP, Pérez-García F, Sgobba E., World J Microbiol Biotechnol 32(6), 2016
PMID: 27116971
Recent advances in amino acid production by microbial cells.
Hirasawa T, Shimizu H., Curr Opin Biotechnol 42(), 2016
PMID: 27151315
Screening of an Escherichia coli promoter library for a phenylalanine biosensor.
Mahr R, von Boeselager RF, Wiechert J, Frunzke J., Appl Microbiol Biotechnol 100(15), 2016
PMID: 27170323
Comparative Single-Cell Analysis of Different E. coli Expression Systems during Microfluidic Cultivation.
Binder D, Probst C, Grünberger A, Hilgers F, Loeschcke A, Jaeger KE, Kohlheyer D, Drepper T., PLoS One 11(8), 2016
PMID: 27525986
Image-Based Single Cell Profiling: High-Throughput Processing of Mother Machine Experiments.
Sachs CC, Grünberger A, Helfrich S, Probst C, Wiechert W, Kohlheyer D, Nöh K., PLoS One 11(9), 2016
PMID: 27661996
Light-Controlled Cell Factories: Employing Photocaged Isopropyl-β-d-Thiogalactopyranoside for Light-Mediated Optimization of lac Promoter-Based Gene Expression and (+)-Valencene Biosynthesis in Corynebacterium glutamicum.
Binder D, Frohwitter J, Mahr R, Bier C, Grünberger A, Loeschcke A, Peters-Wendisch P, Kohlheyer D, Pietruszka J, Frunzke J, Jaeger KE, Wendisch VF, Drepper T., Appl Environ Microbiol 82(20), 2016
PMID: 27520809
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
Modeling and CFD simulation of nutrient distribution in picoliter bioreactors for bacterial growth studies on single-cell level.
Westerwalbesloh C, Grünberger A, Stute B, Weber S, Wiechert W, Kohlheyer D, von Lieres E., Lab Chip 15(21), 2015
PMID: 26345659
Spatiotemporal microbial single-cell analysis using a high-throughput microfluidics cultivation platform.
Grünberger A, Probst C, Helfrich S, Nanda A, Stute B, Wiechert W, von Lieres E, Nöh K, Frunzke J, Kohlheyer D., Cytometry A 87(12), 2015
PMID: 26348020
Biosensor-driven adaptive laboratory evolution of l-valine production in Corynebacterium glutamicum.
Mahr R, Gätgens C, Gätgens J, Polen T, Kalinowski J, Frunzke J., Metab Eng 32(), 2015
PMID: 26453945
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

52 References

Daten bereitgestellt von Europe PubMed Central.

The role of physiological heterogeneity in microbial population behavior.
Lidstrom ME, Konopka MC., Nat. Chem. Biol. 6(10), 2010
PMID: 20852608
Stochastic switching as a survival strategy in fluctuating environments.
Acar M, Mettetal JT, van Oudenaarden A., Nat. Genet. 40(4), 2008
PMID: 18362885
Bistability, epigenetics, and bet-hedging in bacteria.
Veening JW, Smits WK, Kuipers OP., Annu. Rev. Microbiol. 62(), 2008
PMID: 18537474
Origin and analysis of microbial population heterogeneity in bioprocesses
AUTHOR UNKNOWN, 2010
Experimental methods and modeling techniques for description of cell population heterogeneity.
Fernandes RL, Nierychlo M, Lundin L, Pedersen AE, Puentes Tellez PE, Dutta A, Carlquist M, Bolic A, Schapper D, Brunetti AC, Helmark S, Heins AL, Jensen AD, Nopens I, Rottwitt K, Szita N, van Elsas JD, Nielsen PH, Martinussen J, Sorensen SJ, Lantz AE, Gernaey KV., Biotechnol. Adv. 29(6), 2011
PMID: 21540103
Living with heterogeneities in bioreactors: understanding the effects of environmental gradients on cells.
Lara AR, Galindo E, Ramirez OT, Palomares LA., Mol. Biotechnol. 34(3), 2006
PMID: 17284782
Understanding the bioreactor
AUTHOR UNKNOWN, 2002
Scale-up of microbial processes: impacts, tools and open questions.
Takors R., J. Biotechnol. 160(1-2), 2011
PMID: 22206982
Physiological responses to mixing in large scale bioreactors.
Enfors SO, Jahic M, Rozkov A, Xu B, Hecker M, Jurgen B, Kruger E, Schweder T, Hamer G, O'Beirne D, Noisommit-Rizzi N, Reuss M, Boone L, Hewitt C, McFarlane C, Nienow A, Kovacs T, Tragardh C, Fuchs L, Revstedt J, Friberg PC, Hjertager B, Blomsten G, Skogman H, Hjort S, Hoeks F, Lin HY, Neubauer P, van der Lans R, Luyben K, Vrabel P, Manelius A., J. Biotechnol. 85(2), 2001
PMID: 11165362
Single cell metabolomics
AUTHOR UNKNOWN, 2011
High-throughput metabolic engineering: advances in small-molecule screening and selection.
Dietrich JA, McKee AE, Keasling JD., Annu. Rev. Biochem. 79(), 2010
PMID: 20367033

AUTHOR UNKNOWN, 0
A disposable picolitre bioreactor for cultivation and investigation of industrially relevant bacteria on the single cell level.
Grunberger A, Paczia N, Probst C, Schendzielorz G, Eggeling L, Noack S, Wiechert W, Kohlheyer D., Lab Chip 12(11), 2012
PMID: 22511122
Using movies to analyse gene circuit dynamics in single cells.
Locke JC, Elowitz MB., Nat. Rev. Microbiol. 7(5), 2009
PMID: 19369953

AUTHOR UNKNOWN, 0
Monitoring of population dynamics of Corynebacterium glutamicum by multiparameter flow cytometry.
Neumeyer A, Hubschmann T, Muller S, Frunzke J., Microb Biotechnol 6(2), 2012
PMID: 23279937
Export of L-isoleucine from Corynebacterium glutamicum: a two-gene-encoded member of a new translocator family.
Kennerknecht N, Sahm H, Yen MR, Patek M, Saier Jr MH Jr, Eggeling L., J. Bacteriol. 184(14), 2002
PMID: 12081967
Lrp of Corynebacterium glutamicum controls expression of the brnFE operon encoding the export system for L-methionine and branched-chain amino acids.
Lange C, Mustafi N, Frunzke J, Kennerknecht N, Wessel M, Bott M, Wendisch VF., J. Biotechnol. 158(4), 2011
PMID: 21683740
Characterization of methionine export in Corynebacterium glutamicum.
Trotschel C, Deutenberg D, Bathe B, Burkovski A, Kramer R., J. Bacteriol. 187(11), 2005
PMID: 15901702
The development and application of a single-cell biosensor for the detection of l-methionine and branched-chain amino acids.
Mustafi N, Grunberger A, Kohlheyer D, Bott M, Frunzke J., Metab. Eng. 14(4), 2012
PMID: 22583745
L-valine production with pyruvate dehydrogenase complex-deficient Corynebacterium glutamicum.
Blombach B, Schreiner ME, Holatko J, Bartek T, Oldiges M, Eikmanns BJ., Appl. Environ. Microbiol. 73(7), 2007
PMID: 17293513
Comparative 13C metabolic flux analysis of pyruvate dehydrogenase complex-deficient, L-valine-producing Corynebacterium glutamicum.
Bartek T, Blombach B, Lang S, Eikmanns BJ, Wiechert W, Oldiges M, Noh K, Noack S., Appl. Environ. Microbiol. 77(18), 2011
PMID: 21784914
Linking central metabolism with increased pathway flux: L-valine accumulation by Corynebacterium glutamicum.
Radmacher E, Vaitsikova A, Burger U, Krumbach K, Sahm H, Eggeling L., Appl. Environ. Microbiol. 68(5), 2002
PMID: 11976094
Engineering of Corynebacterium glutamicum for high-yield L-valine production under oxygen deprivation conditions.
Hasegawa S, Suda M, Uematsu K, Natsuma Y, Hiraga K, Jojima T, Inui M, Yukawa H., Appl. Environ. Microbiol. 79(4), 2012
PMID: 23241971
Platform engineering of Corynebacterium glutamicum with reduced pyruvate dehydrogenase complex activity for improved production of L-lysine, L-valine, and 2-ketoisovalerate.
Buchholz J, Schwentner A, Brunnenkan B, Gabris C, Grimm S, Gerstmeir R, Takors R, Eikmanns BJ, Blombach B., Appl. Environ. Microbiol. 79(18), 2013
PMID: 23835179
Corynebacterium glutamicum tailored for high-yield L-valine production.
Blombach B, Schreiner ME, Bartek T, Oldiges M, Eikmanns BJ., Appl. Microbiol. Biotechnol. 79(3), 2008
PMID: 18379776

AUTHOR UNKNOWN, 0

AUTHOR UNKNOWN, 0
A rapidly maturing far-red derivative of DsRed-Express2 for whole-cell labeling
AUTHOR UNKNOWN, 2009

AUTHOR UNKNOWN, 0
Beyond growth rate 0.6: Corynebacterium glutamicum cultivated in highly diluted environments.
Grunberger A, van Ooyen J, Paczia N, Rohe P, Schiendzielorz G, Eggeling L, Wiechert W, Kohlheyer D, Noack S., Biotechnol. Bioeng. 110(1), 2012
PMID: 22890752
Green fluorescent protein (GFP) leakage from microbial biosensors provides useful information for the evaluation of the scale-down effect.
Delvigne F, Brognaux A, Francis F, Twizere JC, Gorret N, Sorensen SJ, Thonart P., Biotechnol J 6(8), 2011
PMID: 21695786
Physiological heterogeneities in microbial populations and implications for physical stress tolerance.
Carlquist M, Fernandes RL, Helmark S, Heins AL, Lundin L, Sorensen SJ, Gernaey KV, Lantz AE., Microb. Cell Fact. 11(), 2012
PMID: 22799461

AUTHOR UNKNOWN, 0
Efficient aerobic succinate production from glucose in minimal medium with Corynebacterium glutamicum.
Litsanov B, Kabus A, Brocker M, Bott M., Microb Biotechnol 5(1), 2011
PMID: 22018023
ENZYME INDUCTION AS AN ALL-OR-NONE PHENOMENON.
Novick A, Weiner M., Proc. Natl. Acad. Sci. U.S.A. 43(7), 1957
PMID: 16590055
Multistability in the lactose utilization network of Escherichia coli.
Ozbudak EM, Thattai M, Lim HN, Shraiman BI, Van Oudenaarden A., Nature 427(6976), 2004
PMID: 14973486
A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level.
Binder S, Schendzielorz G, Stabler N, Krumbach K, Hoffmann K, Bott M, Eggeling L., Genome Biol. 13(5), 2012
PMID: 22640862
Biosensors and their applications in microbial metabolic engineering.
Zhang F, Keasling J., Trends Microbiol. 19(7), 2011
PMID: 21664818
Applications of genetically-encoded biosensors for the construction and control of biosynthetic pathways.
Michener JK, Thodey K, Liang JC, Smolke CD., Metab. Eng. 14(3), 2011
PMID: 21946159
Studies on the amino acid fermentation: I. Production of L-glutamic acid by various microorganisms
AUTHOR UNKNOWN, 1957

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