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@ARTICLE{Seide:906840,
author = {Seide, Selina and Arnold, Lilia and Wetzels, Solange and
Bregu, Mariela and Gätgens, Jochem and Pohl, Martina},
title = {{F}rom {E}nzyme to {P}reparative {C}ascade {R}eactions with
{I}mmobilized {E}nzymes: {T}uning
{F}e({II})/α-{K}etoglutarate-{D}ependent {L}ysine
{H}ydroxylases for {A}pplication in {B}iotransformations},
journal = {Catalysts},
volume = {12},
number = {4},
issn = {2073-4344},
address = {Basel},
publisher = {MDPI},
reportid = {FZJ-2022-01734},
pages = {354 -},
year = {2022},
abstract = {Fe(II)/α-ketoglutarate-dependent dioxygenases (KDOs)
catalyze a broad range of selective C–H oxidation
reactions. However, the difficult production of KDOs in
recombinant E. coli strains and their instability in
purified form have so far limited their application in
preparative biotransformations. Here, we investigated the
immobilization of three KDOs (CaKDO, CpKDO, FjKDO) that
catalyze the stereoselective hydroxylation of the L-lysine
side chain using two one-step immobilization techniques
(HaloTag®, EziG™). The HaloTag®-based immobilisates
reached the best results with respect to residual activity
and stability. In preparative lab-scale experiments, we
achieved product titers of 16 g L−1 (3S)-hydroxy-L-lysine
(CaKDO) and (4R)-hydroxy-L-lysine (FjKDO), respectively,
starting from 100 mM L-lysine. Using a HaloTag®-immobilized
lysine decarboxylase from Selenomonas ruminantium (SrLDC),
the (3S)-hydroxy-L-lysine from the CaKDO-catalyzed reaction
was successfully converted to (2S)-hydroxy-cadaverine
without intermediate product purification, yielding a
product titer of 11.6 g L−1 in a 15 mL consecutive batch
reaction. We propose that covalent in situ immobilization is
an appropriate tool to access the preparative potential of
many other KDOs.},
cin = {IBG-1},
ddc = {540},
cid = {I:(DE-Juel1)IBG-1-20101118},
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},
UT = {WOS:000786882300001},
doi = {10.3390/catal12040354},
url = {https://juser.fz-juelich.de/record/906840},
}