JACS Au,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Dec. 17, 2024
Naturally
occurring
photoenzymes
are
rare
in
nature,
but
among
them,
fatty
acid
photodecarboxylases
derived
from
Chlorella
variabilis
(CvFAPs)
have
emerged
as
promising
photobiocatalysts
capable
of
performing
the
redox-neutral,
light-induced
decarboxylation
free
acids
(FAs)
into
C1-shortened
n-alka(e)nes.
Using
a
hybrid
QM/MM
approach
combined
with
polarizable
embedding
scheme,
we
identify
structural
changes
active
site
and
determine
energetic
landscape
forward
electron
transfer
(fET)
FA
substrate
to
excited
flavin
adenine
dinucleotide.
We
obtain
charge-transfer
diradical
structure
where
water
molecule
rearranges
spontaneously
form
H-bond
interaction
flavin,
while
FA's
carboxylate
group
twists
migrates
away
it.
Together,
these
modifications
provide
driving
force
necessary
for
fET
proceed
downhill
direction.
Moreover,
by
examining
R451K
mutant
is
farther
core,
show
that
marked
reduction
electronic
coupling
counterbalanced
an
increased
force,
resulting
lifetime
similar
WT,
thereby
suggesting
resilience
process
this
mutation.
Finally,
through
molecular
dynamic
simulations,
reveal
that,
following
fET,
radical
occurs
within
tens
picoseconds,
overcoming
energy
barrier
∼0.1
eV.
Overall,
providing
atomistic
characterization
photoactivation
CvFAP,
work
can
be
used
future
protein
engineering.
Journal of the American Chemical Society,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Jan. 23, 2025
Visible
light-driven
pyridoxal
radical
biocatalysis
has
emerged
as
a
new
strategy
for
the
stereoselective
synthesis
of
valuable
noncanonical
amino
acids
in
protecting-group-free
fashion.
In
our
previously
developed
dehydroxylative
C–C
coupling
using
engineered
PLP-dependent
tryptophan
synthases,
an
enzyme-controlled
unusual
α-stereochemistry
reversal
and
pH-controlled
enantiopreference
were
observed.
Herein,
through
high-throughput
photobiocatalysis,
we
evolved
set
stereochemically
complementary
PLP
enzymes,
allowing
both
l-
d-amino
with
enhanced
enantiocontrol
across
broad
pH
window.
These
newly
acid
synthases
permitted
use
range
organoboron
substrates,
including
boronates,
trifluoroborates,
boronic
acids,
excellent
efficiency.
Mechanistic
studies
unveiled
unexpected
racemase
activity
earlier
enzyme
variants.
This
promiscuous
was
abolished
shedding
light
on
origin
enantiocontrol.
Further
mechanistic
investigations
suggest
switch
proton
donor
to
account
stereoinvertive
formation
highlighting
stereoinversion
mechanism
that
is
rare
conventional
two-electron
enzymology.
Science,
Journal Year:
2024,
Volume and Issue:
385(6707), P. 416 - 421
Published: July 25, 2024
Enzymes
capable
of
assimilating
fluorinated
feedstocks
are
scarce.
This
situation
poses
a
challenge
for
the
biosynthesis
compounds
used
in
pharmaceuticals,
agrochemicals,
and
materials.
We
developed
photoenzymatic
hydrofluoroalkylation
that
integrates
motifs
into
olefins.
The
photoinduced
promiscuity
flavin-dependent
ene-reductases
enables
generation
carbon-centered
radicals
from
iodinated
fluoroalkanes,
which
directed
by
photoenzyme
to
engage
enantioselectively
with
approach
facilitates
stereocontrol
through
interaction
between
singular
unit
enzyme,
securing
high
enantioselectivity
at
β,
γ,
or
δ
positions
groups
enzymatic
hydrogen
atom
transfer-a
process
is
notably
challenging
conventional
chemocatalysis.
work
advances
strategies
integrating
chemical
opens
avenues
asymmetric
synthesis
compounds.
Journal of the American Chemical Society,
Journal Year:
2024,
Volume and Issue:
146(32), P. 22476 - 22484
Published: July 4, 2024
Visible
light-driven
pyridoxal
radical
biocatalysis
has
emerged
as
a
promising
strategy
for
the
stereoselective
synthesis
of
valuable
noncanonical
amino
acids
(ncAAs).
Previously,
use
well-tailored
photoredox
catalysts
represented
key
to
enable
efficient
phosphate
(PLP)
enzyme-catalyzed
reactions.
Here,
we
report
PLP-dependent
threonine
aldolase-catalyzed
asymmetric
α-C–H
alkylation
abundant
using
Katritzky
pyridinium
salts
alkylating
agents.
The
engineered
aldolases
allowed
this
redox-neutral
proceed
efficiently,
giving
rise
challenging
α-trisubstituted
and
-tetrasubstituted
ncAA
products
in
protecting-group-free
fashion
with
excellent
enantiocontrol.
Mechanistically,
enantioselective
α-alkylation
capitalizes
on
unique
reactivity
persistent
enzymatic
quinonoid
intermediate
derived
from
PLP
cofactor
acid
substrate
allow
novel
C–C
coupling.
Surprisingly,
photobiocatalytic
process
does
not
require
well-established
operates
through
an
unconventional
photoinduced
generation
involving
PLP-derived
aldimine.
ability
develop
reactions
without
relying
classic
photocatalysts
or
photoenzymes
opens
up
new
avenues
advancing
intermolecular
that
are
known
either
organic
chemistry
enzymology.
ACS Catalysis,
Journal Year:
2025,
Volume and Issue:
15(3), P. 1961 - 1972
Published: Jan. 20, 2025
Despite
the
availability
of
numerous
natural
and
engineered
ene-reductases
(ERs),
enantiocomplementary
synthesis
sterically
hindered
molecules
by
ERs
is
still
limited
poor
substrate
acceptance,
particularly
due
to
insufficient
complementary
stereoselectivity.
Herein,
we
reshaped
substrate-binding
pocket
SeER
from
Saccharomyces
eubayanus
through
semirational
design,
enabling
capable
stereocomplementary
hydrogenating
challenging
substituted
β-cyano
cinnamic
esters.
Compared
wild
type,
variants
exhibited
enhanced
activity
(up
161-fold)
catalytic
efficiency
kcat/KM
358-fold),
displaying
potential
in
synthesizing
various
chiral
esters
with
high
stereoselectivity
99%
ee).
Molecular
dynamics
simulations
demonstrated
that
key
for
superior
performance
well-tuned
pocket,
which
strengthens
stabilizes
recognition.
Furthermore,
elucidated
practicality
asymmetric
GABA
derivatives
(e.g.,
Phenibut,
Baclofen,
Tolibut)
via
chemo-enzymatic
cascade
reactions.
Chemistry - A European Journal,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 17, 2025
Saturated
bioisosteres
of
ortho-substituted
benzenes
are
significant
interest
due
to
their
enhanced
pharmacokinetic
properties,
such
as
improved
metabolic
stability
and
reduced
toxicity,
making
them
valuable
in
drug
design
development.
However,
efficient
synthesis
remains
a
challenge
organic
chemistry.
Herein,
we
report
the
biocatalytic
saturated
using
engineered
artificial
photoenzymes.
The
photoenzyme,
incorporating
genetically
encoded
unnatural
amino
acids
with
benzophenone
photosensitizer
residue,
facilitate
formation
chiral
moderate
enantiomeric
excess
via
energy
transfer
process.
Our
results
demonstrate
versatility
photoenzymes
mediating
new-to-nature
reactions
that
difficult
achieve
conventional
chemical
or
enzymatic
methods.
Journal of the American Chemical Society,
Journal Year:
2025,
Volume and Issue:
unknown
Published: April 1, 2025
Despite
its
synthetic
potential,
catalytic
enantioselective
Smiles
rearrangement
has
remained
elusive.
Through
the
directed
evolution
of
P450
radical
aryl
migratases
(P450Smiles's),
we
describe
first
example
rearrangement.
A
range
racemic
N-arylsulfonyl-α-chloroamides
could
be
transformed
by
P450Smiles
in
an
enantioconvergent
manner,
affording
acyclic
amide
products
possessing
all-carbon
quaternary
stereocenter
with
excellent
chemo-
and
enantioselectivity.
Both
electron-rich
electron-deficient
substituents
were
compatible
migrating
group,
demonstrating
this
P450-catalyzed
is
insensitive
to
electronic
properties
group.
Importantly,
our
evolved
variants
capable
overriding
innate
cyclization
activity
N-alkyl
amidyl
intermediate,
allowing
chemoselective
reductive
formation
products.
Classical
molecular
dynamics
(MD)
simulations
revealed
unusual
enzyme-controlled
chemoselectivity
stems
from
restricted
conformation
within
enzyme
active
site,
disfavoring
pathway.
This
new-to-nature
biocatalytic
asymmetric
showcases
potential
enzymatic
enantioselectivity
control
over
highly
reactive
intermediates
eluding
small-molecule
catalysts.