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@ARTICLE{Oeggl:858538,
author = {Oeggl, Reinhard and Neumann, Timo and Gätgens, Jochem and
Romano, Diego and Noack, Stephan and Rother, Dörte},
title = {{C}itrate as {C}ost-{E}fficient {NADPH} {R}egenerating
{A}gent},
journal = {Frontiers in Bioengineering and Biotechnology},
volume = {6},
issn = {2296-4185},
address = {Lausanne},
publisher = {Frontiers Media},
reportid = {FZJ-2018-07409},
pages = {196},
year = {2018},
abstract = {The economically efficient utilization of NADPH or
NADH-dependent enzymes requires the regeneration of consumed
reduction equivalents. This cofactor regeneration is
classically done via an additional substrate, and if
necessary enzyme. We now demonstrate an easy-to-apply
cofactor regeneration approach, which can especially be used
in screening applications. Simply by applying citrate to a
buffer or directly using citrate/-phosphate buffer NADPH can
be regenerated by native enzymes of the TCA cycle,
practically present in all aerobic living organisms. Apart
from viable-culturable cells, this regeneration approach can
also be applied with lyophilized cells and even crude cell
extracts. This is exemplarily shown for the synthesis of 1
phenylethanol from acetophenone with several
oxidoreductases. The mechanism of NADPH regeneration by TCA
cycle enzymes was further investigated by a transient
isotopic labeling experiment feeding [1,5-13C]citrate. This
revealed that the regeneration mechanism can further be
optimized by genetic modification of two competing internal
citrate metabolization pathways, the glyoxylate shunt and
the glutamate dehydrogenase.},
cin = {IBG-1},
ddc = {570},
cid = {I:(DE-Juel1)IBG-1-20101118},
pnm = {581 - Biotechnology (POF3-581)},
pid = {G:(DE-HGF)POF3-581},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:30631764},
UT = {WOS:000454341200001},
doi = {10.3389/fbioe.2018.00196},
url = {https://juser.fz-juelich.de/record/858538},
}