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@ARTICLE{Otten:865570,
      author       = {Otten, Julia and Tenhaef, Niklas and Jansen, Roman P. and
                      Döbber, Johannes and Jungbluth, Lisa and Noack, Stephan and
                      Oldiges, Marco and Wiechert, Wolfgang and Pohl, Martina},
      title        = {{A} {FRET}-based biosensor for the quantification of
                      glucose in culture supernatants of m{L} scale microbial
                      cultivations},
      journal      = {Microbial cell factories},
      volume       = {18},
      number       = {1},
      issn         = {1475-2859},
      address      = {London},
      publisher    = {Biomed Central},
      reportid     = {FZJ-2019-04935},
      pages        = {143},
      year         = {2019},
      abstract     = {BackgroundIn most microbial cultivations d-glucose is the
                      main carbon and energy source. However, quantification of
                      d-glucose especially in small scale is still challenging.
                      Therefore, we developed a FRET-based glucose biosensor,
                      which can be applied in microbioreactor-based cultivations.
                      This sensor consists of a glucose binding protein sandwiched
                      between two fluorescent proteins, constituting a FRET pair.
                      Upon d-glucose binding the sensor undergoes a conformational
                      change which is translated into a FRET-ratio
                      change.ResultsThe selected sensor shows an apparent Kd below
                      1.5 mM d-glucose and a very high sensitivity of up to $70\%$
                      FRET-ratio change between the unbound and the
                      glucose-saturated state. The soluble sensor was successfully
                      applied online to monitor the glucose concentration in an
                      Escherichia coli culture. Additionally, this sensor was
                      utilized in an at-line process for a Corynebacterium
                      glutamicum culture as an example for a process with
                      cell-specific background (e.g. autofluorescence) and
                      medium-induced quenching. Immobilization of the sensor via
                      HaloTag® enabled purification and covalent immobilization
                      in one step and increased the stability during application,
                      significantly.ConclusionA FRET-based glucose sensor was used
                      to quantify d-glucose consumption in microtiter plate based
                      cultivations. To the best of our knowledge, this is the
                      first method reported for online quantification of d-glucose
                      in microtiter plate based cultivations. In comparison to
                      d-glucose analysis via an enzymatic assay and HPLC, the
                      sensor performed equally well, but enabled much faster
                      measurements, which allowed to speed up microbial strain
                      development significantly.},
      cin          = {IBG-1 / ICS-6},
      ddc          = {570},
      cid          = {I:(DE-Juel1)IBG-1-20101118 / I:(DE-Juel1)ICS-6-20110106},
      pnm          = {581 - Biotechnology (POF3-581)},
      pid          = {G:(DE-HGF)POF3-581},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:31434564},
      UT           = {WOS:000483292300003},
      doi          = {10.1186/s12934-019-1193-y},
      url          = {https://juser.fz-juelich.de/record/865570},
}