ChemCatChem,
Journal Year:
2024,
Volume and Issue:
16(11)
Published: Feb. 14, 2024
Abstract
Alcohol
dehydrogenases
(ADHs)
have
garnered
recognition
for
their
potential
in
the
synthesis
of
pivotal
pharmaceutical
compounds.
However,
utilization
context
piperidone
remains
an
area
ripe
exploration.
In
this
study,
we
examine
performance
alcohol
dehydrogenase
derived
from
Corynebacterium
glutamicum
(
Cg
ADH)
using
a
substrate
analogue
functional
screening
(SAFS)
method
to
elucidate
its
specificity.
To
improve
catalytic
activity
ADH,
phenylalanine/alanine‐scanning
and
iterative
saturation
mutation
(PAS‐ISM)
was
used.
The
most
active
variant,
I151F/I195A,
exhibited
remarkable
10.6‐fold
increase
compared
wild‐type.
Structural
analysis
revealed
that
introduction
residues
151F
195A
led
remodeling
substrate‐binding
pocket,
enabling
additional
p‐π
hydrophobic
interactions
with
substrate,
ultimately
promoting
more
favorable
binding
pose.
This
study
introduces
SAFS
method,
which
enables
identification
enzymes
no
sequence
homology
known
enzymes.
Furthermore,
application
PAS‐ISM
presents
efficient
approach
engineering
dehydrogenases.
These
findings
open
up
promising
avenues
expand
utility
ADHs
piperidone,
thereby
advancing
field
chemistry.
Organic & Biomolecular Chemistry,
Journal Year:
2024,
Volume and Issue:
22(18), P. 3559 - 3583
Published: Jan. 1, 2024
This
review
highlights
the
recent
advancements
in
enzymatic
modifications
of
steroid
scaffolds,
emphasizing
hydroxylation,
ketoreduction,
dehydrogenation,
cascade
reactions,
and
other
modifications.
Advanced Science,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 17, 2025
Abstract
Adipic
acid
(AA),
a
key
aliphatic
dicarboxylic
acid,
is
conventionally
manufactured
through
energy‐intensive,
multi‐step
chemical
processes
with
significant
environmental
impacts.
In
contrast,
biological
production
methods
offer
more
sustainable
alternatives
but
are
often
limited
by
low
productivity.
To
overcome
these
challenges,
this
study
reports
the
engineering
of
single
Escherichia
coli
for
efficient
biosynthesis
AA
starting
from
cyclohexanol
(CHOL),
KA
oil
(mixture
CHOL
and
cyclohexanone
(CHONE)),
or
cyclohexane
(CH).
start
with,
comprehensive
screening
rate‐limiting
enzymes
conducted,
particularly
focusing
on
cytochrome
P450
monooxygenase,
followed
optimization
protein
expression
using
strategies
such
as
fusion,
promoter
replacement,
genome
editing.
Consequently,
an
engineered
E.
capable
efficiently
converting
either
CH
into
obtained,
achieving
remarkable
product
titers
110
22.6
g
L
−1
,
respectively.
This
represents
highest
productivity
record
to
date.
Finally,
developed
biocatalytic
system
successfully
employed
convert
different
cycloalkanes
cycloalkanols
varied
carbon
chain
lengths
their
corresponding
acids,
highlighting
its
great
potential
well
broad
applicability
industrial
applications.
Journal of Agricultural and Food Chemistry,
Journal Year:
2025,
Volume and Issue:
unknown
Published: March 20, 2025
P450
hydroxylation
is
widely
used
in
steroidal
drug
synthesis,
which
can
not
only
activate
the
oxidation
of
inert
C–H
bonds
but
also
increase
pharmacological
activity
compounds
after
modification.
The
two-step
ordered
selective
steroids
catalyzed
by
dihydroxylases
often
forms
key
precursors
or
intermediates
steroid
drugs.
However,
poor
catalytic
performance
dihydroxylase
and
imbalance
lead
to
accumulation
intermediates,
restricts
efficient
synthesis
dihydroxyl
products.
In
this
work,
sequence
(first
at
C7
then
C15)
DHEA
7α,15α-diOH-DHEA
CYP-cl3
from
Colletotrichum
lini
ST-1
was
revealed
with
aid
computational
analysis.
To
obtain
highly
C7α-
C15α-selective
enzymes,
we
designed
a
three-step
modification
strategy
that
includes
high-throughput
screening
C7-hydroxylation
mutants,
semirational
design
C15-hydroxylation
selectivity,
combination
dominant
mutation
sites.
As
result,
successfully
obtained
quadruple
mutant
A83P/E264I/V281A/T315P,
whose
proportion
reached
99.9%.
mechanism
amino
acid
residues
improving
molecular
docking
dynamics
simulation
Our
study
guides
improvement
other
hydroxylase.
Organic Letters,
Journal Year:
2025,
Volume and Issue:
unknown
Published: April 17, 2025
CsCYP21A,
a
steroid
21-hydroxylase
from
Bufo
bufo
gargarizans,
exhibits
unprecedented
sequential
oxidations.
Optimizing
Pichia
pastoris
biotransformation
conditions
enhanced
C21-hydroxylation
selectivity,
converting
14
substrates
to
21-hydroxylated
products,
with
10
conversions
of
>80%
and
4
yields
>80%.
Hydrocortisone
production
reached
1.5
g
L-1
day-1
100
g/L
wet
biomass.
CsCYP21A's
versatility
enables
integration
into
the
synthesis
over
steroidal
drugs,
offering
sustainable
biocatalytic
platform
for
green
pharmaceutical
manufacturing.
Advanced Synthesis & Catalysis,
Journal Year:
2023,
Volume and Issue:
365(23), P. 4024 - 4048
Published: May 17, 2023
Abstract
Since
their
structural
elucidation
in
1935,
the
introduction
and
substitution
of
functional
groups
modification
steroidal
scaffolds
have
been
a
fertile
ground
research
for
synthetic
medicinal
chemists.
The
discovery
steroids
with
hormonal
pharmacological
activity
has
stimulated
tremendous
efforts
to
development
highly
selective
efficient
procedures.
Despite
progress
made,
steroid
chemistry
remains
challenging
preparation
compounds
pharmaceutical
interests
clinical
practice,
often
requires
long
elaborated
synthesis.
In
recent
years,
new
impetus
field
came
advent
enabling
chemical
technologies,
such
as
continuous
flow
chemistry,
which
are
exploited
overcome
problems
that
arise
from
batch
Although
it
is
still
niche
sector,
use
technology
synthesis
functionalization
holds
premise
empower
methodology
provide
innovative
tactics
also
many
hitherto
uncharted
chemistries.
this
review,
scientific
contributions
reported
discussed
terms
set‐up
advantages
offered
concerning
process
efficiency,
optimization,
waste
minimization,
safety
improvement,
easy
scale‐up
costs.
We
highlight
main
challenges,
key
improvements
future
trajectory
application
its
implementation
different
disciplines
photochemistry
biocatalysis
ultimate
goal
streamlining
ACS Catalysis,
Journal Year:
2023,
Volume and Issue:
13(19), P. 13111 - 13116
Published: Sept. 25, 2023
Dehydronandrolone
acetate
(3)
is
a
crucial
precursor
for
the
synthesis
of
C7-functionalized
steroidal
drugs.
However,
current
production
method
involves
laborious
and
environmentally
unfriendly
five-step
chemical
process,
resulting
in
low
efficiency.
To
overcome
this,
we
report
chemoenzymatic
strategy,
involving
one-pot
biocatalytic
C7β-hydroxylation/C17β-ketoreduction
19-norandrostenedione
(1)
by
combination
P450
monooxygenase
17-ketosteroid
reductase
to
generate
C7β-hydroxynandrolone
(2)
as
an
intermediate,
followed
dehydration
esterification
form
3.
Impressively,
gram-scale
3
was
achieved
with
93%
isolated
yield,
which
outperforms
traditional
approach
(68%
yield),
thereby
signaling
great
potential
industrial
applications.
European Journal of Organic Chemistry,
Journal Year:
2023,
Volume and Issue:
27(5)
Published: Dec. 12, 2023
Abstract
The
synthesis
of
steroidal
products
has
garnered
sustained
interest
due
to
their
versatile
bioactivities
and
physicochemical
properties
in
pharmaceutics.
However,
the
complicated
structure
steroids,
especially
highly
oxygenated
skeleton,
presented
significant
challenges.
Benefitted
from
recent
advancements
bioinformatics,
genetics,
enzyme
engineering,
chemoenzymatic
strategy
combines
flexibility
chemical
with
high
regio‐
stereo‐selectivity
biocatalysis,
being
an
effective
approach
for
concise
natural
products.
In
this
concept,
we
review
discuss
advances
approaches
efficient
steroids
analogs,
as
well
future
research
opportunities
ACS Catalysis,
Journal Year:
2023,
Volume and Issue:
13(24), P. 15948 - 15955
Published: Nov. 28, 2023
Chiral
γ-halohydrins
and
β-haloallyl
alcohols
are
important
building
blocks
for
the
synthesis
of
pharmacologically
active
compounds.
Direct
enantioselective
C–H
bond
hydroxylation
halohydrocarbons
is
an
appealing
method
these
Herein,
P450DA
mutants,
which
could
improve
or
reverse
enantioselectivity,
were
generated
by
structure-guided
directed
evolution
based
on
X-ray
crystal
structure
P450DA-M3.
It
catalyzed
benzylic
allylic
with
regio-,
chemo-,
enantioselectivity
provided
desirable
enantiomers
both
chiral
(43–94%
ee)
(79–96%
ee),
while
halogen
atoms
C═C
bonds
in
molecule
remained
unreacted.
This
enzymatic
platform
represents
example
catalytic
systems
achieving
enantiodivergent
control
via
protein
engineering.
