Molecular modification of a GH84 β-N-acetylglucosaminidase from Streptomyces violascens for synthesis of lacto-N-triose II using whey powder and chitin-derived N-acetyl chitobiose
Food Chemistry,
Год журнала:
2025,
Номер
474, С. 143046 - 143046
Опубликована: Янв. 24, 2025
Язык: Английский
Efficient Bioconversion of Crystalline α-Chitin to N-Acetyl-D-glucosamine Driven by a Novel β-N-Acetylglucosaminidase and the Promotion Mechanism Analysis
Journal of Agricultural and Food Chemistry,
Год журнала:
2025,
Номер
unknown
Опубликована: Июнь 2, 2025
Whole-enzyme
cascade
degradation
of
crystalline
α-chitin
is
a
long-term
pursuit,
which
has
been
hampered
by
low
transformation
efficiency
and
heterogeneous
products.
Here,
novel
β-N-acetylglucosaminidase
(NAGase)
from
Sulfuriroseicoccus
oceanibius,
designated
Sohex,
was
biochemically
characterized.
The
enzyme
demonstrated
high
hydrolytic
activity
toward
N-acetyl
chitooligomers
(N-acetyl
COSs)
exhibited
promotive
effects
on
degradation.
Furthermore,
whole-enzyme
system
based
lytic
polysaccharide
monooxygenases
(LPMOs),
chitinases,
Sohex
developed
for
N-acetyl-D-glucosamine
(GlcNAc)
production.
This
achieved
maximum
conversion
rate
45.93%,
with
GlcNAc
purity
reaching
99.99%.
addition
significantly
enhanced
efficiency,
achieving
rates
1.37-fold
2.46-fold
compared
to
the
LPMOs
+
chitinases
sole
system,
respectively.
Moreover,
promotion
mechanisms
in
conjunction
were
elucidated,
making
promising
strategy
converting
into
high-purity
GlcNAc.
Язык: Английский
Engineered β1-3-N-Acetylglucosaminyltransferase Facilitating the One-Pot Multienzyme Synthesis of Human Milk Oligosaccharides
Journal of Agricultural and Food Chemistry,
Год журнала:
2024,
Номер
72(50), С. 28019 - 28027
Опубликована: Дек. 6, 2024
β1-3-linked
N-acetylglucosaminide
is
a
prevalent
carbohydrate
motif
found
in
oligosaccharides,
polysaccharides,
glycoproteins,
and
glycolipids.
It
crucial
component
of
human
milk
oligosaccharides
(HMOs).
Neisseria
meningitidis
β1-3-N-acetylglucosaminyltransferase
(NmLgtA)
catalyzes
the
formation
glycosidic
bond
has
potential
for
use
synthesizing
HMOs.
However,
this
application
hindered
by
challenges
such
as
low
levels
enzyme
expression,
poor
stability,
significant
aggregation.
Since
there
no
available
crystal
structure
NmLgtA,
we
used
its
AlphaFold
2
predicted
to
identify
unfavorable
factors.
We
then
modified
removing
17
N-terminal
amino
acids
substituting
nine
specific
residues.
The
engineered
NmLgtA-Opti
exhibited
improved
thermal
increased
soluble
protein
complete
relief
from
aggregation,
enhanced
catalysis
while
maintaining
catalytic
specificity
substrate
promiscuity.
Furthermore,
maximizes
utilization
can
be
employed
sequential
one-pot
multienzyme
platform
high-yield
production
Язык: Английский