Frontiers in Chemical Biology,
Год журнала:
2024,
Номер
3
Опубликована: Июль 23, 2024
Introduction:
Biocatalysis,
particularly
through
engineered
enzymes,
presents
a
cost-effective,
efficient,
and
eco-friendly
approach
to
compound
synthesis.
We
sought
identify
ketoreductases
capable
of
synthesizing
optically
pure
alcohols
or
ketones,
essential
chiral
building
blocks
for
active
pharmaceutical
ingredients.
Methods:
Using
BioMatchMaker®,
an
in
silico
high-throughput
platform
that
allows
the
identification
wild-type
enzyme
sequences
desired
chemical
transformation,
we
identified
bacterial
SDR
ketoreductase
from
Thermus
caliditerrae,
Tcalid
SDR,
demonstrates
favorable
reaction
efficiency
enantiomeric
excess.
Results:
Here
present
two
crystal
structures
apo-form
at
1.9
Å
NADP-complexed
form
1.7
resolution
(9FE6
9FEB,
respectively).
This
forms
homotetramer
with
each
subunit
containing
N-terminal
Rossmann-fold
domain.
use
computational
analysis
combined
site-directed
mutagenesis
enzymatic
characterization
define
substrate-binding
pocket.
Furthermore,
retained
reactivity
selectivity
after
incubation
elevated
temperature.
Conclusion:
The
enantioselectivity
thermostability
makes
this
attractive
engineering
starting
point
biocatalysis
applications.
Angewandte Chemie International Edition,
Год журнала:
2024,
Номер
63(22)
Опубликована: Апрель 1, 2024
Abstract
The
design
and
orderly
layered
co‐immobilization
of
multiple
enzymes
on
resin
particles
remain
challenging.
In
this
study,
the
SpyTag/SpyCatcher
binding
pair
was
fused
to
N‐terminus
an
alcohol
dehydrogenase
(ADH)
aldo‐keto
reductase
(AKR),
respectively.
A
non‐canonical
amino
acid
(ncAA),
p
‐azido‐L‐phenylalanine
(p‐AzF),
as
anchor
for
covalent
bonding
enzymes,
genetically
inserted
into
preselected
sites
in
AKR
ADH.
Employing
two
bioorthogonal
counterparts
azide–alkyne
cycloaddition
immobilization
ADH
enabled
sequential
dual‐enzyme
coating
porous
microspheres.
ordered
reactor
subsequently
used
synthesize
(
S
)‐1‐(2‐chlorophenyl)ethanol
asymmetrically
from
corresponding
prochiral
ketone,
enabling
situ
regeneration
NADPH.
exhibited
a
high
catalytic
conversion
74
%
good
reproducibility,
retaining
80
its
initial
activity
after
six
cycles.
product
had
99.9
ee,
which
that
maintained
each
cycle.
Additionally,
double‐layer
method
significantly
increased
enzyme
loading
capacity,
approximately
1.7
times
greater
than
traditional
single‐layer
immobilization.
More
importantly,
it
simultaneously
both
purification
carriers,
thus
providing
convenient
approach
facilitate
cascade
biocatalysis.
EES Catalysis,
Год журнала:
2023,
Номер
2(1), С. 14 - 48
Опубликована: Окт. 28, 2023
This
article
reviews
the
integration
of
multidisciplinary
approaches,
including
protein
engineering,
computational
biology,
and
nanoarchitectonics,
to
advance
pharmaceutical
enzyme
biocatalysis.
Green Chemistry,
Год журнала:
2024,
Номер
26(4), С. 2124 - 2134
Опубликована: Янв. 1, 2024
An
asymmetric
sustainable
route
to
d
-pantothenic
acid
was
developed,
centering
on
the
reduction
of
ethyl
2′-ketopantothenate
(
R
)-pantothenate,
enabled
by
an
E.
coli
strain
co-expressing
engineered
ketoreductase
and
glucose
dehydrogenase.
Angewandte Chemie International Edition,
Год журнала:
2023,
Номер
63(13)
Опубликована: Ноя. 4, 2023
The
asymmetric
reduction
of
double
bonds
using
NAD(P)H-dependent
oxidoreductases
has
proven
to
be
an
efficient
tool
for
the
synthesis
important
chiral
molecules
in
research
and
on
industrial
scale.
These
enzymes
are
commercially
available
screening
kits
C=O
(ketones),
C=C
(activated
alkenes),
or
C=N
(imines).
Recent
reports,
however,
indicate
that
ability
accommodate
multiple
reductase
activities
distinct
C=X
occurs
different
enzyme
classes,
either
natively
after
mutagenesis.
This
challenges
common
perception
highly
selective
one
type
electrophilic
substrate.
Consideration
this
underexplored
potential
screenings
protein
engineering
campaigns
may
contribute
identification
complementary
biocatalytic
processes
compounds.
review
will
a
global
understanding
promiscuous
behavior
bond
inspire
future
discoveries
with
respect
unconventional
routes
synthesis.
Chemistry - A European Journal,
Год журнала:
2024,
Номер
unknown
Опубликована: Сен. 6, 2024
Abstract
The
development
of
sustainable
chemistry
underlying
the
quest
to
minimize
and/or
valorize
waste
in
carbon‐neutral
manufacture
chemicals
is
followed
over
last
four
five
decades.
Both
chemo‐
and
biocatalysis
have
played
an
indispensable
role
this
odyssey.
particular
developments
protein
engineering,
metagenomics
bioinformatics
preceding
three
decades
a
crucial
supporting
facilitating
widespread
application
both
whole
cell
cell‐free
biocatalysis.
pressing
need,
driven
by
climate
change
mitigation,
for
drastic
reduction
greenhouse
gas
(GHG)
emissions,
has
precipitated
energy
transition
based
on
decarbonization
defossilization
organic
production.
latter
involves
biomass
CO
2
as
feedstock
green
electricity
generated
using
solar,
wind,
hydroelectric
or
nuclear
energy.
use
polysaccharides
feedstocks
will
underpin
renaissance
carbohydrate
with
pentoses
hexoses
base
bio‐based
solvents
polymers
environmentally
friendly
downstream
products.
availability
inexpensive
solar
led
increasing
attention
electro(bio)catalysis
photo(bio)catalysis
which
turn
leading
myriad
innovations
these
fields.
