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@ARTICLE{Lamm:889648,
      author       = {Lamm, Robin and Jäger, Vera D. and Heyman, Benedikt and
                      Berg, Christoph and Cürten, Christin and Krauss, Ulrich and
                      Jaeger, Karl-Erich and Büchs, Jochen},
      title        = {{D}etailed small-scale characterization and scale-up of
                      active {YFP} inclusion body production with {E}scherichia
                      coli induced by a tetrameric coiled coil domain},
      journal      = {Journal of bioscience and bioengineering},
      volume       = {129},
      number       = {6},
      issn         = {1389-1723},
      address      = {[S.l.]},
      publisher    = {Science Direct},
      reportid     = {FZJ-2021-00280},
      pages        = {730 - 740},
      year         = {2020},
      abstract     = {During heterologous protein production with Escherichia
                      coli, the formation of inclusion bodies (IBs) is often a
                      major drawback as these aggregated proteins are usually
                      inactive. However, different strategies for the generation
                      of IBs consisting of catalytically active proteins have
                      recently been described. In this study, the archaeal
                      tetrameric coiled-coil domain of the cell-surface protein
                      tetrabrachion was fused to a target reporter protein to
                      produce fluorescent IBs (FIBs). As the cultivation
                      conditions severely influence IB formation, the entire
                      cultivation process resulting in the production of FIBs were
                      thoroughly studied. First, the cultivation process was
                      scaled down based on the maximum oxygen transfer capacity,
                      combining online monitoring technologies for shake flasks
                      and microtiter plates with offline sampling. The evaluation
                      of culture conditions in complex terrific broth
                      autoinduction medium showed strong oxygen limitation and
                      leaky expression. Furthermore, strong acetate formation and
                      pH changes from 6.5 to 8.8 led to sub-optimal cultivation
                      conditions. However, in minimal Wilms-MOPS autoinduction
                      medium, defined culture conditions and a tightly controlled
                      expression were achieved. The production of FIBs is strongly
                      influenced by the induction strength. Increasing induction
                      strengths result in lower total amounts of functional
                      protein. However, the amount of functional FIBs increases.
                      Furthermore, to prevent the formation of conventional
                      inactive IBs, a temperature shift from 37 °C to 15 °C is
                      crucial to generate FIBs. Finally, the gained insights were
                      transferred to a stirred tank reactor batch fermentation.
                      Hereby, 12 g/L FIBs were produced, making up 43 $\%$ (w/w)
                      of the total generated biomass.},
      cin          = {IMET / IBT-1},
      ddc          = {660},
      cid          = {I:(DE-Juel1)IMET-20090612 / I:(DE-Juel1)VDB55},
      pnm          = {581 - Biotechnology (POF3-581)},
      pid          = {G:(DE-HGF)POF3-581},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {32143998},
      UT           = {WOS:000614233200012},
      doi          = {10.1016/j.jbiosc.2020.02.003},
      url          = {https://juser.fz-juelich.de/record/889648},
}