bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2023,
Номер
unknown
Опубликована: Ноя. 28, 2023
ABSTRACT
Fluorinated
amino
acids
are
a
promising
entry
point
for
incorporating
new-to-Nature
chemistries
in
biological
systems.
Hence,
novel
methods
needed
the
selective
synthesis
of
these
building
blocks.
In
this
study,
we
focused
on
enzymatic
fluorinated
alanine
enantiomers.
To
end,
dehydrogenase
from
Vibrio
proteolyticus
and
diaminopimelate
Symbiobacterium
thermophilum
were
applied
to
vitro
production
(
R
)-3-fluoroalanine
S
)-3-fluoroalanine,
respectively,
using
3-fluoropyruvate
as
substrate.
Additionally,
an
racemase
Streptomyces
lavendulae
,
originally
selected
setting
alternative
cascade
leading
non-canonical
acids,
had
unprecedented
catalytic
efficiency
β-elimination
fluorine
monosubstituted
fluoroalanine.
The
based
dehydrogenases
V
.
included
cofactor
recycling
system,
whereby
formate
Pseudomonas
sp.
101
(either
native
or
engineered)
coupled
oxidation
NAD(P)H
formation.
Under
conditions,
reaction
yields
reached
>85%
substrate
proceeded
with
complete
enantiomeric
excess.
Moreover,
also
able
catalyze
conversion
trifluoropyruvate
into
trifluorinated
alanine,
first-case
example
biocatalysis
carrying
trifluoromethyl
group.
Current Opinion in Biotechnology,
Год журнала:
2023,
Номер
85, С. 103025 - 103025
Опубликована: Дек. 7, 2023
The
soil
bacterium
Pseudomonas
putida,
especially
the
KT2440
strain,
is
increasingly
being
utilized
as
a
host
for
biotransformations
of
both
industrial
and
environmental
interest.
foundations
such
performance
include
its
robust
redox
metabolism,
ability
to
tolerate
wide
range
physicochemical
stresses,
rapid
growth,
versatile
nonpathogenic
nature,
availability
molecular
tools
advanced
genetic
programming.
These
attributes
have
been
leveraged
hosting
engineered
pathways
production
valuable
chemicals
or
degradation/valorization
pollutants.
This
has
in
turn
pushed
boundaries
conventional
enzymology
toward
previously
unexplored
reactions
nature.
Furthermore,
modifications
physical
properties
cells
made
enhance
their
catalytic
performance.
advancements
establish
P.
putida
bona
fide
chassis
synthetic
biology,
on
par
with
more
traditional
metabolic
engineering
platforms.
FEBS Journal,
Год журнала:
2023,
Номер
290(20), С. 4966 - 4983
Опубликована: Июль 12, 2023
Fluorine
forms
the
strongest
single
bond
to
carbon
with
highest
dissociation
energy
among
natural
products.
However,
fluoroacetate
dehalogenases
(FADs)
have
been
shown
hydrolyze
this
in
under
mild
reaction
conditions.
Furthermore,
two
recent
studies
demonstrated
that
FAD
RPA1163
from
Rhodopseudomonas
palustris
can
also
accept
bulkier
substrates.
In
study,
we
explored
substrate
promiscuity
of
microbial
FADs
and
their
ability
defluorinate
polyfluorinated
organic
acids.
Enzymatic
screening
eight
purified
reported
defluorination
activity
revealed
significant
hydrolytic
against
difluoroacetate
three
proteins.
Product
analysis
using
liquid
chromatography-mass
spectrometry
identified
glyoxylic
acid
as
final
product
enzymatic
DFA
defluorination.
The
crystal
structures
DAR3835
Dechloromonas
aromatica
NOS0089
Nostoc
sp.
were
determined
apo-state
along
H274N
glycolyl
intermediate.
Structure-based
site-directed
mutagenesis
a
key
role
for
catalytic
triad
other
active
site
residues
both
difluoroacetate.
Computational
dimer
DAR3835,
NOS0089,
indicated
presence
one
access
tunnel
each
protomer.
Moreover,
protein-ligand
docking
simulations
suggested
similar
mechanisms
difluoroacetate,
being
defluorinated
via
consecutive
reactions
producing
glyoxylate
product.
Thus,
our
findings
provide
molecular
insights
into
mechanism
FADs,
which
are
promising
biocatalysts
applications
synthetic
chemistry
bioremediation
fluorochemicals.
Communications Chemistry,
Год журнала:
2024,
Номер
7(1)
Опубликована: Май 9, 2024
Abstract
Fluorinated
amino
acids
serve
as
an
entry
point
for
establishing
new-to-Nature
chemistries
in
biological
systems,
and
novel
methods
are
needed
the
selective
synthesis
of
these
building
blocks.
In
this
study,
we
focused
on
enzymatic
fluorinated
alanine
enantiomers
to
expand
fluorine
biocatalysis.
The
dehydrogenase
from
Vibrio
proteolyticus
diaminopimelate
Symbiobacterium
thermophilum
were
selected
vitro
production
(
R
)-3-fluoroalanine
S
)-3-fluoroalanine,
respectively,
using
3-fluoropyruvate
substrate.
Additionally,
discovered
that
racemase
Streptomyces
lavendulae
,
originally
setting
alternative
cascade
leading
non-canonical
acids,
had
unprecedented
catalytic
efficiency
β-elimination
monosubstituted
fluoroalanine.
based
dehydrogenases
V
.
included
a
cofactor
recycling
system,
whereby
formate
Pseudomonas
sp.
101
(either
native
or
engineered)
coupled
oxidation
NAD(P)H
formation.
Under
conditions,
reaction
yields
reached
>85%
substrate
proceeded
with
complete
enantiomeric
excess.
also
catalyzed
conversion
trifluoropyruvate
into
trifluorinated
first-case
example
biocatalysis
carrying
trifluoromethyl
group.
Abstract
From
12
to
14
March,
a
gathering
of
microbiologists
and
biotechnologists
convened
at
the
International
University
Andalucía
(UNIA)
in
Baeza
(Jaén,
Spain)
under
auspices
UNIA
Applied
Microbiology
(AMI)
Society.
The
primary
objective
this
meeting
was
analyse
pivotal
role
microbes
play
sustaining
our
planet
broader
context—both
from
descriptive
(what
is
state
affairs)
prescriptive
perspective
do
look
for
regarding
activities
interest).
workshop
focused
on
exploring
ecological
aspects
soil
water,
including
some
extreme
environments,
potential
microorganisms
as
promoters
plant
growth
biocontrol
agents
well
active
large-scale
catalysts
environmental
health.
Various
topics
were
examined
context,
encompassing
application
platforms
biosynthesis
value-added
chemicals,
bioremediation
technologies,
concept
circular
economy,
emergence
zoonotic
concerns
within
dynamically
changing
global
environment,
precision
nutrition
medicine.
In
alignment
with
strategic
plan
AMI,
discussion
background
United
Nations
Sustainability
Development
Goals
(UN
SDGs).
These
targets
formulated
beginning
past
decade
guide
humanity
towards
sustainable
future.
UN
SDGs
aim
prevent
overstepping
planetary
boundaries,
which
are
endangering
Earth’s
biodiversity
even
human
survival.
iScience,
Год журнала:
2023,
Номер
26(12), С. 108383 - 108383
Опубликована: Ноя. 3, 2023
Major
advances
in
mastering
metabolism
of
single
carbon
(C1)
gaseous
feedstocks
acetogenic
microorganisms
are
primed
to
fuel
the
transition
toward
environmentally
sustainable
and
cost-efficient
production
schemes
biofuels
value-added
biochemicals.
Since
acetogens
grow
under
autotrophic
energy-limited
conditions,
protein
synthesis
is
expected
be
controlled.
This
survey
integrated
publicly
available
RNA
sequencing
ribosome
profiling
studies
several
acetogens,
providing
data
on
genome-scale
transcriptional
translational
responses
A.
woodii,
E.
limosum,
C.
drakei,
ljungdahlii
heterotrophic
growth
conditions.
The
extent
efficiency
turned
out
vary
across
key
functional
modules
acetogens'
metabolism.
Translational
control
was
confirmed
support
stoichiometric
multimeric
complexes.
Comparing
condition
revealed
growth-dependent
regulation
efficiency,
pointing
at
buffering
as
a
widespread
phenomenon
shared
by
acetogens.
