% IMPORTANT: The following is UTF-8 encoded.  This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.

@ARTICLE{Gerlach:906758,
      author       = {Gerlach, Tim and Schain, Jendrik and Söltl, Simone and van
                      Schie, Morten M. C. H. and Hilgers, Fabienne and
                      Bitzenhofer, Nora L. and Drepper, Thomas and Rother, Dörte},
      title        = {{P}hoto-{R}egulation of {E}nzyme {A}ctivity: {T}he
                      {I}nactivation of a {C}arboligase with {G}enetically
                      {E}ncoded {P}hotosensitizer {F}usion {T}ags},
      journal      = {Frontiers in catalysis},
      volume       = {2},
      issn         = {2673-7841},
      address      = {Lausanne},
      publisher    = {Frontiers Media},
      reportid     = {FZJ-2022-01673},
      pages        = {835919},
      year         = {2022},
      abstract     = {Genetically encoded photosensitizers are able to produce
                      reactive oxygen species upon illumination and are exploited
                      in a wide range of applications, especially in the medical
                      field. In this work, we envisioned to further apply these
                      genetically encoded photosensitizers for the light-dependent
                      control of single enzymes in multi-step biocatalysis. One of
                      the challenges in the application of several enzymes in a
                      cascade is the unwanted cross-reactivity of these
                      biocatalysts on reaction intermediates when all enzymes are
                      simultaneously present in the reaction. As one strategy to
                      address this issue, we investigated whether the introduction
                      of genetically encoded photosensitizers as fusion tags would
                      allow the selective inactivation of enzymes after successful
                      transformation by simply turning on light. We tested five
                      different photosensitizers as molecular biological fusion
                      tags to inactivate the pyruvate decarboxylase variant
                      E469G/W543H from Acetobacter pasteurianus. Dimeric
                      photosensitizer tags, like the flavin-binding fluorescent
                      proteins from Bacillus subtilis and Pseudomonas putida
                      showed the tendency to form insoluble protein aggregates in
                      combination with the tetrameric carboligase. Enzyme activity
                      was, to some extent, retained in these aggregates, but the
                      handling of the insoluble aggregates proved to be
                      unfeasible. Monomeric photosensitizer tags appeared to be
                      much more suitable when fused to the tetrameric enzyme. In
                      the dark, the singlet oxygen photosensitizing protein
                      (SOPP3)-tagged carboligase retained $79\%$ of its activity
                      as compared to the unfused enzyme. Upon blue light exposure,
                      the SOPP3 tag showed the best specific inactivation and
                      enabled complete inactivation of the carboligase within 30
                      min. SOPP3 is thus seen as a promising photosensitizer tag
                      to be applied in future multi-step enzyme cascades to
                      overcome the challenge of cross-reactivity.},
      cin          = {IBG-1 / IMET},
      ddc          = {540},
      cid          = {I:(DE-Juel1)IBG-1-20101118 / I:(DE-Juel1)IMET-20090612},
      pnm          = {2172 - Utilization of renewable carbon and energy sources
                      and engineering of ecosystem functions (POF4-217)},
      pid          = {G:(DE-HGF)POF4-2172},
      typ          = {PUB:(DE-HGF)16},
      doi          = {10.3389/fctls.2022.835919},
      url          = {https://juser.fz-juelich.de/record/906758},
}