% 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{Ksters:890631,
      author       = {Küsters, Kira and Pohl, Martina and Krauss, Ulrich and
                      Ölçücü, Gizem and Albert, Sandor and Jaeger, Karl-Erich
                      and Wiechert, Wolfgang and Oldiges, Marco},
      title        = {{C}onstruction and comprehensive characterization of an
                      {E}c{LDC}c-{C}at{IB} set - varying linkers and aggregation
                      inducing tags},
      journal      = {Microbial cell factories},
      volume       = {20},
      number       = {1},
      issn         = {1475-2859},
      address      = {London},
      publisher    = {Biomed Central},
      reportid     = {FZJ-2021-01086},
      pages        = {49},
      year         = {2021},
      abstract     = {BackgroundIn recent years, the production of inclusion
                      bodies that retained substantial catalytic activity was
                      demonstrated. These catalytically active inclusion bodies
                      (CatIBs) were formed by genetic fusion of an aggregation
                      inducing tag to a gene of interest via short linker
                      polypeptides and overproduction of the resulting gene fusion
                      in Escherichia coli. The resulting CatIBs are known for
                      their high stability, easy and cost efficient production,
                      and recyclability and thus provide an interesting
                      alternative to conventionally immobilized
                      enzymes.ResultsHere, we present the construction and
                      characterization of a CatIB set of the lysine decarboxylase
                      from Escherichia coli (EcLDCc), constructed via Golden Gate
                      Assembly. A total of ten EcLDCc variants consisting of
                      combinations of two linker and five aggregation inducing tag
                      sequences were generated. A flexible Serine/Glycine (SG)- as
                      well as a rigid Proline/Threonine (PT)-Linker were tested in
                      combination with the artificial peptides (18AWT, L6KD and
                      GFIL8) or the coiled-coil domains (TDoT and 3HAMP) as
                      aggregation inducing tags. The linkers were fused to the
                      C-terminus of the EcLDCc to form a linkage between the
                      enzyme and the aggregation inducing tags. Comprehensive
                      morphology and enzymatic activity analyses were performed
                      for the ten EcLDCc-CatIB variants and a wild type EcLDCc
                      control to identify the CatIB variant with the highest
                      activity for the decarboxylation of l-lysine to
                      1,5-diaminopentane. Interestingly, all of the CatIB variants
                      possessed at least some activity, whilst most of the
                      combinations with the rigid PT-Linker showed the highest
                      conversion rates. EcLDCc-PT-L6KD was identified as the best
                      of all variants allowing a volumetric productivity of
                      457 g L− 1 d− 1 and a specific volumetric
                      productivity of 256 g L− 1 d− 1 gCatIB−1.
                      Noteworthy, wild type EcLDCc, without specific aggregation
                      inducing tags, also partially formed CatIBs, which, however
                      showed lower activity compared to most of the newly
                      constructed CatIB variants (volumetric productivity: 219 g
                      L− 1 d− 1, specific volumetric activity: 106 g
                      L− 1 d− 1 gCatIB− 1). Furthermore, we demonstrate
                      that microscopic analysis can serve as a tool to find CatIB
                      producing strains and thus allow for prescreening at an
                      early stage to save time and resources.ConclusionsOur
                      results clearly show that the choice of linker and
                      aggregation inducing tag has a strong influence on the
                      morphology and the enzymatic activity of the CatIBs.
                      Strikingly, the linker had the most pronounced influence on
                      these characteristics.},
      cin          = {IBG-1 / IMET},
      ddc          = {570},
      cid          = {I:(DE-Juel1)IBG-1-20101118 / I:(DE-Juel1)IMET-20090612},
      pnm          = {2171 - Biological and environmental resources for
                      sustainable use (POF4-217)},
      pid          = {G:(DE-HGF)POF4-2171},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {33596923},
      UT           = {WOS:000620983800004},
      doi          = {10.1186/s12934-021-01539-w},
      url          = {https://juser.fz-juelich.de/record/890631},
}