Heterologous Expression and Functional Analysis of Exiguobacterium Algin Lyase Gene by Pichia pastoris
Fermentation,
Год журнала:
2025,
Номер
11(1), С. 34 - 34
Опубликована: Янв. 16, 2025
Algin
is
the
most
abundant
substance
in
alga.
Alginate
lyase
degrades
algin
and
produces
monosaccharides,
disaccharides,
oligosaccharides,
which
are
widely
used
bioenergy,
food,
medicine,
other
fields.
In
this
study,
one
Exiguobacterium
strain
isolated
from
rotten
kelp
exhibited
a
robust
ability
to
degrade
The
sequencing
of
revealed
presence
three
different
types
alginate
lyase.
Nevertheless,
expression
genes
Escherichia
coli
lower
activity
compared
that
original
strain.
After
codon
optimization,
gene
with
highest
was
successfully
expressed
Pichia
pastoris
produce
recombinant
EbAlg664.
enzyme
5
L
high-density
fermentation
reached
1306
U/mg
protein,
3.9
times
results
enzymatic
analysis
optimal
temperature
pH
range
EbAlg664
were
narrower
Additionally,
Cu2+
Co2+
enhanced
activity,
whereas
Mg2+
Fe3+
inhibitory
effects
on
study
offers
theoretical
practical
foundation
for
industrial-scale
production
engineered
high
activity.
Язык: Английский
Molecular Matchmakers: How ATP and Small Amphiphilic Molecules Fine-Tune FET Proteins Clusters
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2025,
Номер
unknown
Опубликована: Апрель 16, 2025
Abstract
FET
(FUS-EWSR1-TAF15)
family
proteins
inherently
form
mesoscale
molecular
assemblies,
known
as
clusters,
under
physiological
conditions
at
concentrations
well
below
the
threshold
for
phase
separation.
This
study
demonstrates
that
adenosine
triphosphate
(ATP),
an
amphiphilic
molecule
and
essential
cellular
metabolite,
modulates
size
of
these
sub-saturation
clusters
in
a
concentration-dependent
manner.
At
low
(1-2
mM),
ATP
acts
crosslinker
proteins,
resulting
larger
clusters.
moderate
(5
decreases
but
stabilizes.
high
(10
cluster
further
diminishes.
Other
molecules,
including
common
hydrotropes
like
sodium
xylene
sulfonate,
toluene
hexanediol,
exhibit
comparable
effects
on
protein
clustering.
Notably,
cannot
be
explained
solely
by
hydrotropic
or
kosmotropic
mechanisms;
instead,
they
stem
from
non-specific
interactions
between
small
molecules.
The
intrinsic
chemical
properties
molecules
play
crucial
role
regulating
formation
concentrations.
Язык: Английский
Magnesium: Der Schlüssel zur Langlebigkeit
Zeitschrift für Orthomolekulare Medizin,
Год журнала:
2025,
Номер
23(01), С. 22 - 29
Опубликована: Апрель 1, 2025
Metal-organic frameworks (MOF-74) show distinct bio-effects to nitrogen-fixing bacterium Azotobacter vinelandii
Journal of Hazardous Materials,
Год журнала:
2025,
Номер
494, С. 138779 - 138779
Опубликована: Май 29, 2025
Язык: Английский
Allostery can convert binding free energies into concerted domain motions in enzymes
Nature Communications,
Год журнала:
2024,
Номер
15(1)
Опубликована: Ноя. 22, 2024
Abstract
Enzymatic
mechanisms
are
typically
inferred
from
structural
data.
However,
understanding
enzymes
require
unravelling
the
intricate
dynamic
interplay
between
dynamics,
conformational
substates,
and
multiple
protein
structures.
Here,
we
use
single-molecule
nanopore
analysis
to
investigate
catalytic
changes
of
adenylate
kinase
(AK),
an
enzyme
that
catalyzes
interconversion
various
adenosine
phosphates
(ATP,
ADP,
AMP).
Kinetic
validated
by
hidden
Markov
models
unravels
details
domain
motions
during
catalysis.
Our
findings
reveal
allosteric
interactions
ligands
cofactor
enable
converting
binding
energies
into
directional
two
domains
AK.
These
coordinated
emerged
control
exact
sequence
ligand
affinity
for
three
different
substrates,
thereby
guiding
reactants
along
reaction
coordinates.
Interestingly,
find
about
10%
show
altered
regulation
affinities,
indicating
a
subset
folds
in
alternative
catalytically
active
forms.
Since
molecules
or
proteins
might
be
able
selectively
stabilize
one
folds,
this
observation
suggests
evolutionary
path
allostery
enzymes.
In
AK,
complex
framework
has
likely
prevent
futile
ATP/ADP
hydrolysis
regulate
energy
needs
cell.
Язык: Английский