The development and application of a single-cell biosensor for the detection of l-methionine and branched-chain amino acids

Mustafi N, Grünberger A, Kohlheyer D, Bott M, Frunzke J (2012)
Metabolic Engineering 14(4): 449-457.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Mustafi, Nurije; Grünberger, AlexanderUniBi; Kohlheyer, Dietrich; Bott, Michael; Frunzke, Julia
Abstract / Bemerkung
The detection and quantification of specific metabolites in single bacterial cells is a major goal for industrial biotechnology. We have developed a biosensor based on the transcriptional regulator Lrp that detects intracellular l-methionine and branched-chain amino acids in Corynebacterium glutamicum. In assays, fluorescence output showed a linear relationship with cytoplasmic concentrations of the effector amino acids. In increasing order, the affinity of Lrp for the amino acids is l-valine, l-isoleucine, l-leucine and l-methionine. The sensor was applied for online monitoring and analysis of cell-to-cell variability of l-valine production by the pyruvate dehydrogenase-deficient C. glutamicum strain ΔaceE. Finally, the sensor system was successfully used in a high-throughput (HT) FACS screen for the isolation of amino acid-producing mutants after random mutagenesis of a non-producing wild type strain. These applications illustrate how one of nature's sensor devices – transcriptional regulators – can be used for the analysis, directed evolution and HT screening for microbial strain development. Highlights ► Development of a single cell biosensor for the intracellular detection of amino acids. ► Biosensor-based online monitoring of valine production of a C. glutamicum strain. ► Visualization of cell-to-cell variability of microbial production strains. ► Establishment of a FACS high-throughput screening.
Corynebacterium glutamicum; Lrp; GFP; amino acids; l-valine
Metabolic Engineering
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Mustafi N, Grünberger A, Kohlheyer D, Bott M, Frunzke J. The development and application of a single-cell biosensor for the detection of l-methionine and branched-chain amino acids. Metabolic Engineering. 2012;14(4):449-457.
Mustafi, N., Grünberger, A., Kohlheyer, D., Bott, M., & Frunzke, J. (2012). The development and application of a single-cell biosensor for the detection of l-methionine and branched-chain amino acids. Metabolic Engineering, 14(4), 449-457. doi:10.1016/j.ymben.2012.02.002
Mustafi, Nurije, Grünberger, Alexander, Kohlheyer, Dietrich, Bott, Michael, and Frunzke, Julia. 2012. “The development and application of a single-cell biosensor for the detection of l-methionine and branched-chain amino acids”. Metabolic Engineering 14 (4): 449-457.
Mustafi, N., Grünberger, A., Kohlheyer, D., Bott, M., and Frunzke, J. (2012). The development and application of a single-cell biosensor for the detection of l-methionine and branched-chain amino acids. Metabolic Engineering 14, 449-457.
Mustafi, N., et al., 2012. The development and application of a single-cell biosensor for the detection of l-methionine and branched-chain amino acids. Metabolic Engineering, 14(4), p 449-457.
N. Mustafi, et al., “The development and application of a single-cell biosensor for the detection of l-methionine and branched-chain amino acids”, Metabolic Engineering, vol. 14, 2012, pp. 449-457.
Mustafi, N., Grünberger, A., Kohlheyer, D., Bott, M., Frunzke, J.: The development and application of a single-cell biosensor for the detection of l-methionine and branched-chain amino acids. Metabolic Engineering. 14, 449-457 (2012).
Mustafi, Nurije, Grünberger, Alexander, Kohlheyer, Dietrich, Bott, Michael, and Frunzke, Julia. “The development and application of a single-cell biosensor for the detection of l-methionine and branched-chain amino acids”. Metabolic Engineering 14.4 (2012): 449-457.

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