ACS Catalysis,
Journal Year:
2024,
Volume and Issue:
14(21), P. 15988 - 15996
Published: Oct. 15, 2024
Flavoenzymes
are
versatile
biocatalysts
with
extraordinary
properties.
They
not
only
vital
to
life
by
orchestrating
complex
transformations
in
biological
processes
but
offer
numerous
biocatalytic
opportunities.
Most
flavoenzymes
have
a
noncovalently
bound
flavin
cofactor.
The
cofactor
is
responsible
for
the
catalytic
versatility
and
prowess
of
essential
their
structure
stability.
dissociation
adenine
dinucleotide
(FAD)
results
an
immediate
loss
activity
often
unstable
apoenzyme,
which
reduces
turnover
stability
limits
broader
application
industry.
Herein,
we
present
proof-of-concept
light-induced
covalent
flavinylation
three
glucose
oxidase
(GOx),
cellobiose
dehydrogenase
(CDH),
cyclohexanone
monooxygenase
(CHMO).
We
designed
synthesized
seven
FAD
analogs
bearing
diazirine
group,
allowing
light-triggered
anchoring
One
photoclickable
(diazFAD4)
could
be
integrated
successfully
into
all
interest,
achieving
activities
comparable
those
native
FAD.
bond
formation
was
initiated
ultraviolet
(UV)
light
irradiation
confirmed
via
fluorescence
imaging.
By
expanding
photoaffinity
labeling
technique,
provide
novel
method
functional
incorporation
cofactors
biocatalysts.
ACS Omega,
Journal Year:
2024,
Volume and Issue:
9(47), P. 46698 - 46732
Published: Nov. 14, 2024
Immobilization
of
lipases
by
physical
adsorption
improves
their
stability,
recovery,
and
reusability
in
biotechnological
processes.
The
present
review
provides
an
advanced
bibliometric
analysis
a
comprehensive
overview
research
progress
this
field.
By
searching
Web
Science,
39,575
publications
were
analyzed,
325
relevant
articles
selected.
Key
journals,
countries,
institutions,
authors
identified.
most
cited
focus
on
biofuel
production
industrial
applications.
revealed
four
themes
with
the
biofuel.
method
is
effective
when
appropriate
support
used.
Despite
decrease
patent
applications,
interest
remains
high.
Future
studies
should
optimizing
materials
exploring
new
applications
technique.
detailed
understanding
immobilization
adsorption.
Protein Science,
Journal Year:
2025,
Volume and Issue:
34(2)
Published: Jan. 22, 2025
Abstract
Enzyme
immobilization
is
indispensable
for
enhancing
enzyme
performance
in
various
industrial
applications.
Typically,
enzymes
require
specific
spatial
arrangements
optimal
functionality,
underscoring
the
importance
of
correct
orientation.
Despite
well‐known
N‐
or
C‐terminus
tailoring
techniques,
alternatives
achieving
orientation
control
are
limited.
Here,
we
propose
a
novel
approach
that
tailors
surface
with
engineered
His‐rich
loops.
To
aim,
first
solve
X‐ray
crystal
structure
hexameric
alcohol
dehydrogenase
from
Thermus
thermophilus
HB27
(TtHBDH)
(PDB:
9FBD
).
Guided
by
this
3D
structure,
engineer
new
loop
enriched
six
His
residues
to
Molecular
dynamics
simulations
reveal
loop's
imidazole
rings
have
greater
solvent
accessibility
than
those
native
residues,
allowing
more
efficient
on
certain
metal
chelate‐functionalized
carriers.
Using
carriers
functionalized
iron
(III)‐catechol,
apparent
V
max
immobilized
variant
doubles
His‐tagged
one,
and
vice
versa
when
both
variants
copper
(II)‐imidodiacetic
acid.
loop‐engineered
TtHBDH
show
high
operational
stability
reaching
100%
bioconversion
after
10
reaction
cycles,
yet
faster
one.
Chem & Bio Engineering,
Journal Year:
2025,
Volume and Issue:
unknown
Published: March 17, 2025
The
production
of
active
pharmaceutical
ingredients
(APIs)
requires
enantiopure
chiral
amines,
for
which
greener
synthesis
processes
are
needed.
Transaminases
(TAs)
enzymes
that
catalyze
the
enantioselective
amines
from
prochiral
ketones
through
transamination
under
mild
conditions.
Yet,
industrial
applications
biocatalytic
remain
currently
hindered
by
limited
stability
soluble
and
unfavorable
thermodynamic
equilibrium
targeted
asymmetric
reactions.
Enzyme
immobilization
can
be
applied
to
address
stability,
recoverability,
reusability
issues.
In
perspective
process
intensification,
we
chose
immobilize
TAs
on
polymeric
(polypropylene)
membranes.
(R)-2-fluoro-α-methylbenzylamine
((R)-FMBA),
such
membrane-immobilized
exhibited
superior
specific
activity
compared
with
TAs;
they
also
outperformed
immobilized
resins.
reaction
yield
remained,
however,
thermodynamics.
To
further
enhance
yield,
was
coupled
in
situ
crystallization
(R)-FMBA
3,3-diphenylpropionic
acid
(DPPA).
By
doing
so,
theoretical
conversion
pushed
∼44%
∼83%.
fact,
a
72%
overall
recovery
crystallized
demonstrated.
enantioselectivity
mixture
preserved.
Importantly,
purification
greatly
facilitated
since
target
amine
readily
recovered
as
high-purity
(R)-FMBA:DPPA
crystals.
membranes
were
found
fully
reusable,
performing
successive
high-yield
syntheses
only
minor
deactivation.
Overall,
crystallization-assisted
strategy
proposed
herein
offers
path
valuable
targets.
Journal of the American Chemical Society,
Journal Year:
2024,
Volume and Issue:
146(28), P. 18817 - 18822
Published: July 5, 2024
NAD(H)-dependent
enzymes
play
a
crucial
role
in
the
biosynthesis
of
pharmaceuticals
and
fine
chemicals,
but
limited
recyclability
NAD(H)
cofactor
hinders
its
more
general
application.
Here,
we
report
generation
mechano-responsive
PEI-modified
Cry3Aa
protein
crystals
their
use
for
NADH
recycling
over
multiple
reaction
cycles.
For
demonstration
practical
utility,
complementary
particle
containing
genetically
encoded
co-immobilized
formate
dehydrogenase
regeneration
leucine
catalyzing
NADH-dependent
l-tert-leucine
(l-tert-Leu)
has
been
produced.
When
combined
with
crystal,
resultant
system
could
be
used
efficient
l-tert-Leu
up
to
21
days
10.5-fold
improvement
turnover
number.
Bioresources and Bioprocessing,
Journal Year:
2024,
Volume and Issue:
11(1)
Published: July 18, 2024
Abstract
Reductive
amination
by
amine
dehydrogenases
is
a
green
and
sustainable
process
that
produces
only
water
as
the
by-product.
In
this
study,
continuous
flow
was
designed
utilizing
packed
bed
reactor
filled
with
co-immobilized
dehydrogenase
wh84
glucose
for
highly
efficient
biocatalytic
synthesis
of
chiral
amino
alcohols.
The
immobilized
exhibited
better
thermo-,
pH
solvent
stability
high
activity
recovery.
(
S
)-2-aminobutan-1-ol
produced
in
up
to
99%
conversion
ee
processes,
space-time
yields
were
124.5
g
L
-1
d
.
reactions
also
extended
48
h
affording
91.8%
average
conversions.
This
study
showcased
important
potential
production
alcohols
processes.
The
dynamic
properties
of
enzymatic
reaction
networks
(ERNs)
are
difficult
to
predict
due
the
emergence
allosteric
interactions,
product
inhibitions
and
competition
for
resources,
that
all
only
materialize
once
have
been
assembled.
Combining
experimental
kinetics
studies
with
computational
modelling
allows
us
extract
information
on
these
emergent
build
predictive
models.
Here,
we
utilized
pentose
phosphate
pathway
demonstrate
previously
reported
approaches
construct
maximally
informative
datasets
can
be
significantly
improved
by
pulsing
both
enzymes
substrates
into
microfluidic
flow
reactors
(instead
only).
Our
method
augments
available
from
online
databases,
map
behaviours
a
network.
