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

Metabolic Engineering
Nurije MustafiJulia Frunzke

Abstract

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.

References

Jun 21, 1991·Science·J E Bailey
Sep 1, 1995·Molecular Microbiology·D M SitnikovT O Baldwin
Oct 6, 1998·Annual Review of Biochemistry·R Y Tsien
Aug 17, 2002·Science·Michael B ElowitzPeter S Swain
Feb 3, 2005·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Karen DeuschleWolf B Frommer
May 20, 2005·Journal of Bacteriology·Christian TrötschelReinhard Krämer
Nov 5, 2005·Trends in Biotechnology·Dmitriy M ChudakovKonstantin A Lukyanov
Nov 22, 2005·Nature Methods·Nathan C ShanerRoger Y Tsien
Apr 15, 2006·Science·Ben N G GiepmansRoger Y Tsien
Feb 13, 2007·Applied and Environmental Microbiology·Bastian BlombachBernhard J Eikmanns
Apr 28, 2007·Proceedings of the National Academy of Sciences of the United States of America·Jin Hwan ParkSang Yup Lee
Dec 25, 2007·Applied and Environmental Microbiology·Christine Nicole S Santos, Gregory Stephanopoulos
Apr 2, 2008·Applied Microbiology and Biotechnology·Bastian BlombachBernhard J Eikmanns
May 20, 2008·Journal of Bacteriology·Julia FrunzkeMichael Bott
Apr 7, 2010·Annual Review of Biochemistry·Jeffrey A DietrichJay D Keasling
Jun 2, 2010·Nature Reviews. Microbiology·Jan Roelof van der Meer, Shimshon Belkin
Aug 28, 2010·Journal of Microbiology and Biotechnology·Christoph J BoltenChristoph Wittmann
Oct 12, 2010·Current Opinion in Biotechnology·Matthias Heinemann, Renato Zenobi
Jun 15, 2011·Trends in Microbiology·Fuzhong Zhang, Jay Keasling
Sep 29, 2011·Metabolic Engineering·Joshua K MichenerChristina D Smolke

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Citations

Apr 6, 2013·Journal of Structural Biology·Dawn T EriksenHuimin Zhao
May 13, 2014·Applied Microbiology and Biotechnology·Marco OldigesBastian Blombach
Sep 18, 2013·Journal of Microbiological Methods·Christopher ProbstDietrich Kohlheyer
Nov 9, 2014·Applied Microbiology and Biotechnology·Thomas Willke
Dec 18, 2013·Journal of Visualized Experiments : JoVE·Alexander GruenbergerDietrich Kohlheyer
Jan 13, 2016·Applied Microbiology and Biotechnology·Pengfei GuQingsheng Qi
Feb 2, 2016·ACS Synthetic Biology·Sara CletoTimothy K Lu
Feb 26, 2016·Journal of Molecular Biology·Kevin C MaTimothy K Lu
Jan 13, 2015·Current Opinion in Biotechnology·Lothar EggelingJan Marienhagen
Dec 4, 2015·International Journal of Molecular Sciences·Weihong MinJingsheng Liu
Nov 2, 2015·Applied Microbiology and Biotechnology·Regina Mahr, Julia Frunzke
Feb 10, 2016·Proceedings of the National Academy of Sciences of the United States of America·Jameson K Rogers, George M Church
Dec 4, 2012·Microbial Biotechnology·Stefan WieschalkaBernhard J Eikmanns
Apr 17, 2013·Biotechnology Journal·Zhen Chen, An-Ping Zeng
Jan 12, 2013·Microbial Biotechnology·Miroslav PátekJan Nešvera
Jan 3, 2013·Microbial Biotechnology·Andrea NeumeyerJulia Frunzke
Mar 22, 2016·Nature Chemical Biology·Yi XiaoFuzhong Zhang
Jul 6, 2015·Metabolic Engineering·Di LiuFuzhong Zhang
Oct 21, 2014·Environmental Microbiology·Christian Dusny, Andreas Schmid
Feb 1, 2014·Current Opinion in Biotechnology·Marcus SchallmeyJan Marienhagen
Mar 13, 2015·Applied Microbiology and Biotechnology·Lothar Eggeling, Michael Bott
Jan 31, 2015·Applied Microbiology and Biotechnology·Wenshan Liu, Rongrong Jiang
Apr 28, 2016·World Journal of Microbiology & Biotechnology·Volker F WendischElvira Sgobba
May 7, 2016·Current Opinion in Biotechnology·Takashi Hirasawa, Hiroshi Shimizu
May 14, 2016·Applied Microbiology and Biotechnology·Regina MahrJulia Frunzke
Jun 11, 2016·Current Opinion in Biotechnology·Zhiqing Wang, Patrick C Cirino
Oct 19, 2016·Current Opinion in Microbiology·Christian Rückert
Jan 26, 2017·ACS Synthetic Biology·Yongfei LiuDawei Zhang
Apr 11, 2017·Microbial Biotechnology·Teunke van RossumServé W M Kengen
Jul 10, 2014·Scientific Reports·Artur ReimerJan Roelof van der Meer
Jun 25, 2017·Metabolic Engineering·Dennis BinderAlexander Grünberger
Jun 5, 2014·Integrative Biology : Quantitative Biosciences From Nano to Macro·Dennis BinderThomas Drepper
Mar 21, 2018·SLAS Discovery·Stefan Bröer
Feb 28, 2018·Biotechnology Journal·Feng ChengTsvetan Kardashliev
Mar 30, 2018·Applied Microbiology and Biotechnology·Xiaoyuan WangPeter J Quinn
Feb 25, 2015·Lab on a Chip·Christian DusnyAndreas Schmid
Sep 3, 2017·Applied and Environmental Microbiology·Xin SongSheng Yang

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