Nature Communications,
Год журнала:
2022,
Номер
13(1)
Опубликована: Ноя. 7, 2022
Abstract
Microbial
consortia
have
been
considered
potential
platforms
for
bioprocessing
applications.
However,
the
complexity
in
process
control
owing
to
use
of
multiple
strains
necessitates
an
efficient
population
strategy.
Herein,
we
report
circuit-guided
synthetic
acclimation
as
a
strategy
improve
biochemical
production
by
microbial
consortium.
We
designed
consortium
comprising
alginate-utilizing
Vibrio
sp.
dhg
and
3-hydroxypropionic
acid
(3-HP)-producing
Escherichia
coli
direct
conversion
alginate
3-HP.
introduced
genetic
circuit,
named
“Population
guider”,
E.
strain,
which
degrades
ampicillin
only
when
3-HP
is
produced.
In
presence
selection
pressure,
was
successfully
acclimated
increased
4.3-fold
compared
that
simple
co-culturing
during
48-h
fermentation.
believe
this
concept
useful
development
robust
consortium-based
bioprocesses.
Journal of Agricultural and Food Chemistry,
Год журнала:
2024,
Номер
72(25), С. 14216 - 14228
Опубликована: Июнь 11, 2024
Two-component
systems
(TCSs)
sensing
and
responding
to
various
stimuli
outside
inside
cells
are
valuable
resources
for
developing
biosensors
with
synthetic
biology
applications.
However,
the
use
of
TCS-based
suffers
from
a
limited
effector
spectrum,
hypersensitivity,
low
dynamic
range,
unwanted
signal
crosstalk.
Here,
we
developed
tailor-made
Escherichia
coli
whole-cell
γ-aminobutyric
acid
(GABA)
biosensor
by
engineering
chimeric
GABA
chemoreceptor
PctC
TCS.
By
testing
different
TCSs,
PctC/PhoQ
showed
response
GABA.
Chimera-directed
evolution
introduction
insulated
pair
PctC/PhoQ*PhoP*
produced
up
3.50-fold
range
good
orthogonality.
To
further
enhance
lower
basal
leakage,
three
strategies,
PhoP
DNA
binding
sites,
fine-tuning
reporter
expression
optimizing
transcription/translation
components,
tobacco
etch
virus
protease-controlled
protein
degradation,
were
integrated.
This
displayed
large
(15.8-fold),
high
threshold
level
(22.7
g
L-1).
Finally,
optimized
was
successfully
applied
in
high-throughput
microdroplet
screening
GABA-overproducing
Corynebacterium
glutamicum,
demonstrating
its
desired
properties
extracellular
biosensing.
Metabolic Engineering Communications,
Год журнала:
2024,
Номер
19, С. e00250 - e00250
Опубликована: Ноя. 6, 2024
Transcription
factor-based
biosensors
are
genetic
tools
that
aim
to
predictability
link
the
presence
of
a
specific
input
stimuli
tailored
gene
expression
output.
The
performance
characteristics
biosensor
fundamentally
determines
its
potential
applications.
However,
current
methods
engineer
and
optimise
responses
highly
nonintuitive,
struggle
investigate
multidimensional
sequence/design
space
efficiently.
In
this
study
we
employ
design
experiments
(DoE)
approach
build
framework
for
efficiently
engineering
activator-based
with
performances,
apply
development
polyethylene
terephthalate
(PET)
plastic
degradation
monomer
(TPA).
We
simultaneously
core
promoter
operator
regions
responsive
promoter,
by
employing
dual
refactoring
approach,
were
able
explore
an
enhanced
assign
their
causative
effects.
employed
here
serves
as
foundational
transcriptional
enabled
dynamic
range
diverse
signal
output,
sensitivity,
steepness.
further
demonstrate
applicability
on
primary
screening
PET
hydrolases
enzyme
condition
screening,
demonstrating
statistical
modelling
in
optimising
industrial
environmental
Nucleic Acids Research,
Год журнала:
2022,
Номер
50(6), С. 3535 - 3550
Опубликована: Фев. 20, 2022
Cell-free
systems
that
display
complex
functions
without
using
living
cells
are
emerging
as
new
platforms
to
test
our
understanding
of
biological
well
for
practical
applications
such
biosensors
and
biomanufacturing.
Those
use
cell-free
protein
synthesis
(CFPS)
enable
genetically
programmed
have
relied
on
genetic
regulatory
components
found
or
engineered
in
cells.
However,
constraints
cell
permeability,
metabolic
stability,
toxicity
signaling
molecules
prevent
development
devices
even
if
not
subject
constraints.
Efforts
engineer
directly
CFPS
thus
far
been
based
low-throughput
experimental
approaches,
limiting
the
availability
basic
build
with
diverse
functions.
Here,
we
report
a
high-throughput
screening
method
riboswitches
respond
small
molecules.
Droplet-sorting
riboswitch
variants
system
rapidly
identified
compounds
amenable
bacterial
methods.
Finally,
used
histamine
demonstrate
chemical
communication
between
cell-sized
droplets.
Nature Communications,
Год журнала:
2022,
Номер
13(1)
Опубликована: Ноя. 7, 2022
Abstract
Microbial
consortia
have
been
considered
potential
platforms
for
bioprocessing
applications.
However,
the
complexity
in
process
control
owing
to
use
of
multiple
strains
necessitates
an
efficient
population
strategy.
Herein,
we
report
circuit-guided
synthetic
acclimation
as
a
strategy
improve
biochemical
production
by
microbial
consortium.
We
designed
consortium
comprising
alginate-utilizing
Vibrio
sp.
dhg
and
3-hydroxypropionic
acid
(3-HP)-producing
Escherichia
coli
direct
conversion
alginate
3-HP.
introduced
genetic
circuit,
named
“Population
guider”,
E.
strain,
which
degrades
ampicillin
only
when
3-HP
is
produced.
In
presence
selection
pressure,
was
successfully
acclimated
increased
4.3-fold
compared
that
simple
co-culturing
during
48-h
fermentation.
believe
this
concept
useful
development
robust
consortium-based
bioprocesses.
