% 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{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},
}