Research Square (Research Square),
Journal Year:
2023,
Volume and Issue:
unknown
Published: Oct. 13, 2023
Abstract
Guanosine
is
a
purine
nucleoside
that
widely
used
as
raw
material
for
food
additives
and
pharmaceutical
products.
However,
the
lack
of
strains
with
efficiently
producing
guanosine
greatly
limited
industrial
application.
We
attempted
to
produce
guanosinein
Escherichia
coli
using
systematic
metabolic
engineering.
First,
we
overexpressed
synthesis
pathway
from
Bacillus
subtilis
the
prs
gene,
deleted
three
genes
involved
in
guanosinecatabolism
increase
accumulation.
Subsequently,
attenuated
purA
expression
eliminated
feedback
inhibition.
Then,
modified
flux
glycolysis
Entner-Doudoroff
pathways
performed
redox
cofactors
rebalancing.
Finally,
transporter
engineering
further
increased
titre
123.6
mg/L.
After
72
h
fed-batch
fermentation
shake-flask,
achieved
145.2
Our
results
reveal
guanosinesynthesis
was
successfully
optimized
by
combinatorial
engineering,
which
could
be
applicable
efficient
other
Microbial Cell Factories,
Journal Year:
2024,
Volume and Issue:
23(1)
Published: June 19, 2024
Abstract
Background
Guanosine
is
a
purine
nucleoside
that
widely
used
as
raw
material
for
food
additives
and
pharmaceutical
products.
Microbial
fermentation
the
main
production
method
of
guanosine.
However,
guanosine-producing
strains
possess
multiple
metabolic
pathway
interactions
complex
regulatory
mechanisms.
The
lack
with
efficiently
producing-guanosine
greatly
limited
industrial
application.
Results
We
attempted
to
produce
guanosine
in
Escherichia
coli
using
systematic
engineering.
First,
we
overexpressed
synthesis
from
Bacillus
subtilis
prs
gene,
deleted
three
genes
involved
catabolism
increase
accumulation.
Subsequently,
attenuated
purA
expression
eliminated
feedback
transcription
dual
inhibition.
Then,
modified
flux
glycolysis
Entner-Doudoroff
(ED)
pathways
performed
redox
cofactors
rebalancing.
Finally,
transporter
engineering
enhancing
further
increased
titre
134.9
mg/L.
After
72
h
fed-batch
shake-flask,
achieved
289.8
Conclusions
Our
results
reveal
was
successfully
optimized
by
combinatorial
engineering,
which
could
be
applicable
efficient
other
Applied and Environmental Microbiology,
Journal Year:
2024,
Volume and Issue:
90(4)
Published: March 20, 2024
ABSTRACT
Currently,
the
L-malic
acid
titer
achieved
through
Aspergillus
niger
fermentation
reaches
201
g/L,
meeting
industrial
demands
satisfactorily.
However,
co-presence
of
structurally
similar
fumaric
and
succinic
in
products
suggests
a
theoretical
potential
for
further
improvement
production.
In
tricarboxylic
cycle,
fumarate
reductase
mediates
conversion
to
acid.
Subsequently,
fumarase
catalyzes
Notably,
both
enzymatic
reactions
are
reversible.
Our
investigation
revealed
that
A.
contains
only
one
mitochondria-located
FumA.
Employing
CRISPR-Cas9
technology,
we
performed
replacement
fumA
promoter
with
doxycycline-induced
Tet.
Under
non-inducing
condition,
conditional
strain
exhibited
increased
levels
It
strongly
FumA
mainly
promotes
flow
Furthermore,
PmfsA
is
exclusively
activated
medium
by
calcium
carbonate
was
identified
RNA-sequencing
screening.
Utilizing
regulate
expression
led
9.0%
increase
titer,
an
8.75%
yield
(glucose
acid),
8.86%
enhancement
productivity.
This
research
serves
as
significant
step
toward
expediting
industrialization
synthesis
via
biological
fermentation.
Additionally,
it
offers
valuable
insights
biosynthesis
other
organic
acids.
IMPORTANCE
study
focuses
on
enhancing
modifying
cycle
within
mitochondria
.
We
emphasize
role
converting
into
acid,
our
understanding
metabolic
pathways
The
precise
regulation
highlighted
key
factor
this
introduces
stringent
(PmfsA),
CaCO
3
utilization
resulted
heightened
titers.
progress
engineering
bioprocess
optimization
holds
promise
Moreover,
carries
implications
various
Journal of Agricultural and Food Chemistry,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 18, 2025
Itaconic
acid
(IA)
is
a
multifunctional
platform
chemical
with
numerous
biological
functions.
Here,
Trichoderma
reesei
was
engineered
as
remarkable
cell
factory
to
produce
IA.
Heterologous
overexpression
of
the
mitochondrial
tricarboxylate
transport
protein
and
cis-aconitate
decarboxylase
from
Aspergillus
terreus
in
T.
initiated
IA
production
titer
20
g/L.
By
increasing
copy
number
mttA
cadA
plasma
membrane
transporter
proteins
(MFSA),
reached
56.7
The
precursor
synthesis
pathway
overexpressed
by
aconitase
citrate
synthase,
competition
blocked
deletion
P450
monooxygenase
gene
cyp3
further
enhance
production.
final
strain
resulted
93.5
g/L
through
fed-batch
fermentation
1
L
bioreactor.
Our
study
demonstrates
that
can
serve
relevant
industry
for
Biotechnology Journal,
Journal Year:
2024,
Volume and Issue:
19(7)
Published: July 1, 2024
Traditional
Chinese
food
therapies
often
motivate
the
development
of
modern
medicines,
and
learning
from
them
will
bring
bright
prospects.
Monascus,
a
conventional
fungus
with
centuries
use
in
industry,
produces
various
metabolites,
including
natural
pigments,
lipid-lowering
substances,
other
bioactive
ingredients.
Recent
Monascus
studies
focused
on
metabolite
biosynthesis
mechanisms,
strain
modifications,
fermentation
process
optimizations,
significantly
advancing
lab
scale.
However,
advanced
manufacture
for
is
lacking,
restricting
its
scale
production.
Here,
synthetic
biology
techniques
their
challenges
engineering
filamentous
fungi
were
summarized,
especially
Monascus.
With
further
in-depth
discussions
automatic
solid-state
manufacturing
prospects
combining
intensification,
industrial
production
succeed
help
improvement
intelligent
control,
promoting
applications
food,
cosmetic,
agriculture,
medicine,
environmental
protection
industries.
