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@ARTICLE{Steffen:823847,
      author       = {Steffen, Victoria and Otten, Julia and Engelmann, Susann
                      and Radek, Andreas and Limberg, Michael and König, Bernd
                      and Noack, Stephan and Wiechert, Wolfgang and Pohl, Martina},
      title        = {{A} {T}oolbox of {G}enetically {E}ncoded {FRET}-{B}ased
                      {B}iosensors for {R}apid l-{L}ysine {A}nalysis},
      journal      = {Sensors},
      volume       = {16},
      number       = {10},
      issn         = {1424-8220},
      address      = {Basel},
      publisher    = {MDPI},
      reportid     = {FZJ-2016-06487},
      pages        = {1604},
      year         = {2016},
      abstract     = {Background: The fast development of microbial production
                      strains for basic and fine chemicals is increasingly carried
                      out in small scale cultivation systems to allow for higher
                      throughput. Such parallelized systems create a need for new
                      rapid online detection systems to quantify the respective
                      target compound. In this regard, biosensors, especially
                      genetically encoded Förster resonance energy transfer
                      (FRET)-based biosensors, offer tremendous opportunities. As
                      a proof-of-concept, we have created a toolbox of FRET-based
                      biosensors for the ratiometric determination of L-lysine in
                      fermentation broth. Methods: The sensor toolbox was
                      constructed based on a sensor that consists of an optimized
                      central lysine-/arginine-/ornithine-binding protein (LAO-BP)
                      flanked by two fluorescent proteins (enhanced cyan
                      fluorescent protein (ECFP), Citrine). Further sensor
                      variants with altered affinity and sensitivity were obtained
                      by circular permutation of the binding protein as well as
                      the introduction of flexible and rigid linkers between the
                      fluorescent proteins and the LAO-BP, respectively. Results:
                      The sensor prototype was applied to monitor the
                      extracellular L-lysine concentration of the L-lysine
                      producing Corynebacterium glutamicum (C. glutamicum) strain
                      DM1933 in a BioLector® microscale cultivation device. The
                      results matched well with data obtained by HPLC analysis and
                      the Ninhydrin assay, demonstrating the high potential of
                      FRET-based biosensors for high-throughput microbial
                      bioprocess optimization.},
      cin          = {ICS-6 / IBG-1},
      ddc          = {620},
      cid          = {I:(DE-Juel1)ICS-6-20110106 / I:(DE-Juel1)IBG-1-20101118},
      pnm          = {553 - Physical Basis of Diseases (POF3-553)},
      pid          = {G:(DE-HGF)POF3-553},
      typ          = {PUB:(DE-HGF)16},
      UT           = {WOS:000386131600057},
      doi          = {10.3390/s16101604},
      url          = {https://juser.fz-juelich.de/record/823847},
}