International Journal of Molecular Sciences,
Год журнала:
2023,
Номер
24(22), С. 16067 - 16067
Опубликована: Ноя. 8, 2023
Formate
dehydrogenases
catalyze
the
reversible
oxidation
of
formate
to
carbon
dioxide.
These
enzymes
play
an
important
role
in
CO2
reduction
and
serve
as
nicotinamide
cofactor
recycling
enzymes.
More
recently,
CO2-reducing
activity
dehydrogenases,
especially
metal-containing
has
been
further
explored
for
efficient
atmospheric
capture.
Here,
we
investigate
binding
site
dehydrogenase
from
Rhodobacter
capsulatus
its
specificity
toward
NAD+
vs.
NADP+
reduction.
Starting
NAD+-specific
wild-type
RcFDH,
key
residues
were
exchanged
enable
on
basis
NAD+-bound
cryo-EM
structure
(PDB-ID:
6TG9).
It
observed
that
lysine
at
position
157
(Lys157)
β-subunit
enzyme
is
essential
NAD+.
RcFDH
variants
had
Glu259
either
a
positively
charged
or
uncharged
amino
acid
additional
with
NADP+.
The
FdsBL279R
FdsBK276A
also
showed
Kinetic
parameters
all
determined
tested
able
reduce
using
NADPH
electron
donor
coupled
assay
phosphite
(PTDH),
which
regenerates
NADPH.
This
makes
suitable
applications
where
it
can
be
other
use
Bioresources and Bioprocessing,
Год журнала:
2025,
Номер
12(1)
Опубликована: Май 5, 2025
Abstract
Nicotinamide
cofactor
biomimetics
(NCBs)
serve
as
low-cost
alternatives
to
the
expensive
NAD(P)
+
/NAD(P)H,
holding
significant
potential
for
applications
in
oxidoreductases.
In
this
study,
an
alcohol
dehydrogenase
(
Sp
ADH2)
from
Sphingobium
sp.
SYK-6
was
identified
utilization
of
synthetic
NCBs.
ADH2
exhibited
a
catalytic
activity
11.55
U/g
oxidation
syringyl
when
utilizing
para
-3-carbamoyl-1-(4-carboxybenzyl)pyridin-1-ium
p
-BANA
)
cofactor.
Semi-rational
engineering
led
identification
key
variants
(H43L,
A290I,
H43L/A290I)
with
enhanced
efficiency
and
specificity
using
Compared
wild-type,
variant
H43L/A290I
7-fold
increase
astonishing
6750-fold
improvement
ratio.
Enzymatic
characterization
reveals
that
substrate
spectrum
could
change
significantly
different
totally
NCBs
(tsNCBs).
Furthermore,
interaction
analysis
demonstrates
critical
roles
residues
43
290
anchoring
release
.
This
study
natural
ADH
capable
NCBs,
which
has
never
been
reported.
Importantly,
our
results
provide
valuable
candidates
biology
industrial
developments,
offer
guidance
ADHs
toward
cofactors
improved
performance.
International Journal of Molecular Sciences,
Год журнала:
2023,
Номер
24(22), С. 16067 - 16067
Опубликована: Ноя. 8, 2023
Formate
dehydrogenases
catalyze
the
reversible
oxidation
of
formate
to
carbon
dioxide.
These
enzymes
play
an
important
role
in
CO2
reduction
and
serve
as
nicotinamide
cofactor
recycling
enzymes.
More
recently,
CO2-reducing
activity
dehydrogenases,
especially
metal-containing
has
been
further
explored
for
efficient
atmospheric
capture.
Here,
we
investigate
binding
site
dehydrogenase
from
Rhodobacter
capsulatus
its
specificity
toward
NAD+
vs.
NADP+
reduction.
Starting
NAD+-specific
wild-type
RcFDH,
key
residues
were
exchanged
enable
on
basis
NAD+-bound
cryo-EM
structure
(PDB-ID:
6TG9).
It
observed
that
lysine
at
position
157
(Lys157)
β-subunit
enzyme
is
essential
NAD+.
RcFDH
variants
had
Glu259
either
a
positively
charged
or
uncharged
amino
acid
additional
with
NADP+.
The
FdsBL279R
FdsBK276A
also
showed
Kinetic
parameters
all
determined
tested
able
reduce
using
NADPH
electron
donor
coupled
assay
phosphite
(PTDH),
which
regenerates
NADPH.
This
makes
suitable
applications
where
it
can
be
other
use
