Any feedback?
Information on EC 2.4.1.212 - hyaluronan synthase and Organism(s) Pasteurella multocida and UniProt Accession Q7BLV3
for references in articles please use BRENDA:EC2.4.1.212Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
2.4.1.212 hyaluronan synthaseIUBMB Comments
The enzyme from Streptococcus Group A and Group C requires Mg2+. The enzyme adds GlcNAc to nascent hyaluronan when the non-reducing end is GlcA, but it adds GlcA when the non-reducing end is GlcNAc . The enzyme is highly specific for UDP-GlcNAc and UDP-GlcA; no copolymerization is observed if either is replaced by UDP-Glc, UDP-Gal, UDP-GalNAc or UDP-GalA. Similar enzymes have been found in a variety of organisms.
Specify your search results
Word Map
- 2.4.1.212
- glycosaminoglycans
- collagen
- polysaccharide
- keratinocytes
- cartilage
- 4-methylumbelliferone
- proteoglycans
- hyaluronidases
- chondrocytes
-
dermal
- mesenchymal
-
pericellular
- pasteurella
-
udp-glucuronic
-
hyals
- versican
-
cumulus
- synovial
-
vocal
-
articular
- aggrecan
- osteoarthritis
- streptococcal
- udp-n-acetylglucosamine
- multocida
-
rhamm
- pyogenes
- udp-sugars
- chondroitin
-
glucuronic
- procollagen
- synoviocytes
-
cumulus-oocyte
- decorin
- udp-glcua
-
photoaging
- filaggrin
- equisimilis
-
ha-binding
-
temporomandibular
-
glcua
-
perineuronal
- ophthalmopathy
- zooepidemicus
-
uvb-irradiated
-
ha-mediated
-
tnfaip6
- medicine
-
hyaluronan-binding
- fibromodulin
- biglycan
- analysis
- synthesis
- drug development
- biotechnology
The taxonomic range for the selected organisms is: Pasteurella multocida
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Synonyms
hyaluronan synthase, ha synthase, hyaluronan synthase 2, has-2, hyaluronic acid synthase, sehas, has-1, pmhas, hyaluronan synthase-2, hyaluronate synthase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
CHAS2
-
-
-
-
CHAS3
-
-
-
-
DG42 protein
-
-
-
-
HA synthase
HuHAS1
-
-
-
-
hyaluronan synthethase
-
-
-
-
hyaluronate synthase
-
-
-
-
hyaluronate synthetase
-
-
-
-
hyaluronic acid synthase
-
-
-
-
hyaluronic acid synthetase
-
-
-
-
XHAS1
-
-
-
-
XHAS2
-
-
-
-
XHAS3
-
-
-
-
additional information
-
the enzyme of Pasteurella multocida belongs to the group of class II hyaluronan synthases
HA synthase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
UDP-N-acetyl-alpha-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] = UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
a mechanistic shift from a steady-state ordered bi-bi to rapid equilibrium ordered bi-bi mechanism is observed at the NAc-site between the HA6 and HA8 elongation
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan] = UDP + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan]
a mechanistic shift from a steady-state ordered bi-bi to rapid equilibrium ordered bi-bi mechanism is observed at the NAc-site between the HA6 and HA8 elongation
UDP-N-acetyl-alpha-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] = UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
UDP-N-acetyl-alpha-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] = UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
catalytic mechanism
-
UDP-N-acetyl-alpha-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] = UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
2 active sites, the 2 DXD motifs of the transferase sites in the C-terminus, residues 686-703, are essential for activity
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hexosyl group transfer
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
Alternating UDP-alpha-N-acetyl-D-glucosamine:beta-D-glucuronosyl-(1->3)-[nascent hyaluronan] 4-N-acetyl-beta-D-glucosaminyltransferase and UDP-alpha-D-glucuronate:N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] 3-beta-D-glucuronosyltransferase
The enzyme from Streptococcus Group A and Group C requires Mg2+. The enzyme adds GlcNAc to nascent hyaluronan when the non-reducing end is GlcA, but it adds GlcA when the non-reducing end is GlcNAc [3]. The enzyme is highly specific for UDP-GlcNAc and UDP-GlcA; no copolymerization is observed if either is replaced by UDP-Glc, UDP-Gal, UDP-GalNAc or UDP-GalA. Similar enzymes have been found in a variety of organisms.
CAS REGISTRY NUMBER
COMMENTARY
39346-43-5
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
UDP-alpha-D-glucuronate + hyaluronan oligomer HA5
UDP + beta-D-glucuronosyl-(1->3)-hyaluronan oligomer HA5
-
-
-
?
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-[nascent hyaluronan]
UDP + beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-[nascent hyaluronan]
-
-
-
?
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan]
UDP + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan]
-
-
-
?
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-[nascent hyaluronan]
UDP + N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-[nascent hyaluronan]
-
-
-
?
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
-
-
-
?
UDP-alpha-N-acetyl-D-glucosamine + hyaluronan oligomer HA4
UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-hyaluronan oligomer HA4
-
-
-
?
UDP-alpha-N-acetyl-D-glucosamine + hyaluronan oligomer HA6
UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-hyaluronan oligomer HA6
-
-
-
?
UDP-alpha-N-acetyl-D-glucosamine + hyaluronan oligomer HA8
UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-hyaluronan oligomer HA8
-
-
-
?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
-
-
?
[hyaluronan](n) + UDP-alpha-D-glucuronate
H+ + beta-D-glucuronosyl-(1->4)-[hyaluronan](n) + UDP
-
-
-
?
[hyaluronan](n) + UDP-N-acetyl-alpha-D-glucosamine
H+ + N-acetyl-beta-D-glucosaminyl-(1->4)-[hyaluronan](n) + UDP
-
-
-
?
hyaluronic acid tetrasaccharide + UDP-alpha-N-acetyl-D-glucosamine
?
-
-
-
-
?
UDP-D-glucuronate + chondroitin 4-sulfate trisaccharide
?
-
3.6% of the activity with hyaluronan
-
-
?
UDP-D-glucuronate + chondroitin 6-sulfate pentasaccharide
?
-
61% of the activity with hyaluronan
-
-
?
UDP-D-glucuronate + chondroitin 6-sulfate trisaccharide
?
-
80% of the activity with hyaluronan
-
-
?
UDP-D-glucuronate + chondroitin sulfate
?
-
12% of the activity with hyaluronan
-
-
?
UDP-D-glucuronate + unsulfated chondroitin
?
-
54% of the activity with hyaluronan
-
-
?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
-
-
-
?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
chains growth at the non-reducing end, which is terminated by lack of substrate with a non-reducing end, active with exogenously added acceptors substrates
-
-
?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
formation of linear hyaluronan polymers composed of alternating beta3-N-acetylglucosamine-beta4-glucuronic acid
the product chain length can grow at the reducing end up to 40000 monosaccharides with a MW of over 8 million Da before it is released by the class I enzyme
-
?
additional information
?
-
Pasteurella multocida hyaluronan synthase encompasses two transferase domains that elongate a growing hyaluronan oligosaccharide chain by addition of either GlcNAc or GlcUA residues from a corresponding UDP-sugar
-
-
?
additional information
?
-
a monodispersed hyaluronan chain can be obtained by finely tuning the reaction stoichiometry. The molar ratio of precursors and acceptor molecules has an important role in enzyme kinetics
-
-
?
additional information
?
-
initial velocity studies of single-step elongations are conducted for both domains by independently varying the concentrations of the hyaluronan oligosaccharide and the UDP-sugar. Two-substrate models are discriminated by their goodness-of-fit parameters and by dead-end inhibition studies. Coupled-enzyme assay using LDH, PK; NADH and phosphoenolpyruvate, as well as hyaluronan oligosaccharides, UDP-GlcNAc and UDP-GlcUA, overview
-
-
?
additional information
?
-
-
enzyme is responsible for hyaluronan biosynthesis, the hyaluronan capsule is an important, but not the only, virulence factor, physiological role of the enzyme
-
-
?
additional information
?
-
-
the produced hyaluronan capsule enhances infection
-
-
?
additional information
?
-
-
enzyme is not processive, enzyme requires other proteins for hyaluronan translocation
-
-
?
additional information
?
-
-
substrate specificity, the GlcNAc-transferase, but not the GlcUA-transferase activity depends on the WGGED motif, overview
-
-
?
additional information
?
-
-
PmHAS elongates a range of acceptor molecules in addition to the cognate sugars. Certain glycosaminoglycans are very poor acceptors in comparison to the cognate molecules, but elongated products are detected. The interaction between the acceptor and the enzyme (a) the orientation of the hydroxyl at the C-4 position of the hexosamine is not critical, (b) the conformation of C-5 of the hexuronic acid (glucuronic versus iduronic) is not crucial, and (c) additional negative sulfate groups are well tolerated in certain cases, such as on C-6 of the hexosamine, but others, including C-4 sulfates, are not or are poorly tolerated
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-[nascent hyaluronan]
UDP + beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-[nascent hyaluronan]
-
-
-
?
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan]
UDP + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan]
-
-
-
?
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-[nascent hyaluronan]
UDP + N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-[nascent hyaluronan]
-
-
-
?
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
-
-
-
?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
-
-
?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
-
-
-
?
additional information
?
-
Pasteurella multocida hyaluronan synthase encompasses two transferase domains that elongate a growing hyaluronan oligosaccharide chain by addition of either GlcNAc or GlcUA residues from a corresponding UDP-sugar
-
-
?
additional information
?
-
-
enzyme is responsible for hyaluronan biosynthesis, the hyaluronan capsule is an important, but not the only, virulence factor, physiological role of the enzyme
-
-
?
additional information
?
-
-
the produced hyaluronan capsule enhances infection
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K+
coactivation occurs if K+ is applied together with Mn2+ or Mg2+. With Mn2+, a high UDP reaction rate and high molecular mass of hyaluronic acid (2.45 MDa after 8 h) are achieved. With Mg2+, a low reaction rate and low molecular mass (1.55 MDa after 8 h) are reached. If 10 mM K+ are added to 15 mM Mg2+, a significant increase of the reaction rate by a factor of 2.7 is observed, and the molecular mass is doubled (3.11 MDa after 8 h). The coactivating effect of K+ together with Mn2+ is less pronounced
Mg2+
Mn2+
Co2+
-
18% and 39% of the wild-type GlcUA-transferase and GlcNAc-transferase activity with Mn2+, respectively, 0.2 mM
Mg2+
additional information
-
metal requirements of mutant enzymes, overview
Mg2+
Mg2+ activates with an optimum of 15-20 mM leading to about half of the yield obtained with Mn2+ after 8 h
Mn2+
Mn2+ is the preferred metal ion with an optimum of 10 mM. Presence of Mn2+ induces a concentration-dependent decomposition of UDP-GlcA to its 1,2-cyclic phosphate and UMP, especially promoted at basic pH
Mg2+
-
66% and 77% of the wild-type GlcUA-transferase and GlcNAc-transferase activity with Mn2+, respectively, 0.2 mM
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
UDP-glucuronate
The substrate UDP-GlcA inhibits pmHAS in concentrations above 8 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
cardiolipin
the bacterial enzyme is strictly associated with cardiolipin and the catalytic activity is dependent on such lipid
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.7
hyaluronan oligomer HA4
with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme
-
0.6
hyaluronan oligomer HA5
with UDP-alpha-D-glucuronate, pH 8.0, 35°C, recombinant enzyme
-
1
hyaluronan oligomer HA6
with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme
-
0.7
hyaluronan oligomer HA8
with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme
-
0.8
UDP-alpha-D-glucuronate
pH 7.0, 25 °C, 10 mM MnCl2 and 15 mM UDP-GlcNAc (in absence of HA oligosaccharide as acceptor substrate)
23.4
UDP-N-acetyl-alpha-D-glucosamine
pH 7.0, 25 °C, 10 mM MnCl2 and 5 mM UDP-GlcA (in absence of HA oligosaccharide as acceptor substrate)
additional information
additional information
structural and kinetic analysis and modeling, both NAc- and UA-transferase domains follow a sequential kinetic mechanism, most likely an ordered one in which the UDP-sugar donor binds first, followed by the HA oligosaccharide. After transfer of the sugar moiety, both products are released, first the elongated HA oligosaccharide and then the UDP sugar. A mechanistic shift from a steady-state ordered bi-bi to rapid equilibrium ordered bi-bi mechanism is observed at the NAc-site between the HA6 and HA8 elongation, detailed overview
-
additional information
additional information
-
kinetics, wild-type and mutant enzymes
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
7.9
hyaluronan oligomer HA4
with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme
-
7.9
hyaluronan oligomer HA5
with UDP-alpha-D-glucuronate, pH 8.0, 35°C, recombinant enzyme
-
13.5
hyaluronan oligomer HA6
with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme
-
8.8
hyaluronan oligomer HA8
with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
highest activity in MES, MOPS and HEPES buffers and lower activity in TRIS-buffer
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
the only member of Class 2 HAS, expressed by Pasteurella multocida, has a short C-terminal domain with only 4 transmembrane regions and 2 membrane associated domains, structure comparisons, overview
additional information
-
predicted enzyme topology and organization, overview
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D247E
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D247K
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D247N
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D249E
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D249K
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D249N
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D370E
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low hyaluronan synthase activity
D370K
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity
D370N
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity
D477K
-
mutants possess UDP-N-acetyl-D-glucosamine-transferase activity
D527E
-
site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity
D527K
-
site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity
D527N
-
site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity
D529E
-
site-directed mutagenesis, mutant possesses GlcNAc-transferase activity, and low hyaluronan synthase activity
D529K
-
site-directed mutagenesis, mutant possesses GlcNAc-transferase activity, and very low hyaluronan synthase activity
D529N
-
site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity
E369D
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity
E369H
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity
E369Q
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity
additional information
additional information
-
activity rescue of mutants with high substrate concentrations, overview, deletion of residues 1-117 does not affect polymerization activity, construction of different chimeric mutant enzymes comprising residues from Pasteurella multocida type A enzyme and residues of a Pasteurella multocida type F chondroitin synthase, producing an unsulfated chondroitin capsule, the chimeric mutants show different percentages of hyaluronan and chondroitin synthase ativities, overview
additional information
-
a truncated soluble form of recombinant PmHAS (residues 1703) can catalyze the glycosyl transfers in a time- and concentration-dependent manner
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant soluble C-terminally His6-tagged PmHAS1-703 by nickel affinity chromatography
recombinant synthesis of hyaluronan is carried out with Agrobacterium sp. strain ATCC 31749, hyaluronan is primarly found in the culture medium
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene hyaD, recombinant expression of the soluble C-terminally His6-tagged PmHAS1-703 truncated enzyme from pET101/D-TOPO expression vector with an additional V5 epitope
the Escherichia coli expression vector pQE80L and the broad host range cloning vector pBBR122 are used
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
synthesis
multi-enzyme strategy for the in vitro one-pot synthesis of high-molecular-weight hyaluronic acid from substrates sucrose and N-acetylglucosamine (GlcNAc) with in situ regeneration of nucleotide sugars. With optimized reaction conditions, hyaluronic acid with a molecular mass above 2 MDa is synthesized from catalytic UDP concentrations and 10 mM GlcNAc in less than 10 h
analysis
-
a rapid, continuous, and convenient three-enzyme coupled UV absorption assay is developed to quantitate the glucuronic acid and N-acetylglucosamine transferase activities of hyaluronan synthase. Activity is measured by coupling the UDP produced from the PmHAS-catalyzed transfer of UDP-GlcNAc and UDP-GlcUA to a hyaluronic acid tetrasaccharide primer with the oxidation of NADH. Using a fluorescently labeled primer, the products are characterized by gel electrophoresis. The assay can be used to determine kinetic parameters, inhibition constants, and mechanistic aspects of this enzyme. In addition, it can be used to quantify PmHAS during purification of the enzyme from culture media
synthesis
-
it may be possible to generate compounds that will selectively inhibit the binding of hyaluronan to one particular hyaladherin species without perturbing other species. Such sugar molecules may have future utility as selective therapeutics with minimal side effects for diseases such as cancer, autoimmune disease, inflammation, and infection
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
DeAngelis, P.L.
Molecular directionality of polysaccharide polymerization by the Pasteurella multocida hyaluronan synthase
J. Biol. Chem.
274
26557-26562
1999
Pasteurella multocida (Q7BLV3), Pasteurella multocida
Jing, W.; DeAngelis, P.L.
Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide
Glycobiology
10
883-889
2000
Pasteurella multocida
Jing, W.; DeAngelis, P.L.
Analysis of the two active sites of the hyaluronan synthase and the chondroitin synthase of Pasteurella multocida
Glycobiology
13
661-671
2003
Pasteurella multocida, Pasteurella multocida type A
Weigel, P.H.
Functional characteristics and catalytic mechanisms of the bacterial hyaluronan synthases
IUBMB Life
54
201-211
2002
Pasteurella multocida, Streptococcus pyogenes
Krupa, J.C.; Shaya, D.; Chi, L.; Linhardt, R.J.; Cygler, M.; Withers, S.G.; Mort, J.S.
Quantitative continuous assay for hyaluronan synthase
Anal. Biochem.
361
218-225
2007
Pasteurella multocida
Pummill, P.E.; Kane, T.A.; Kempner, E.S.; DeAngelis, P.L.
The functional molecular mass of the Pasteurella hyaluronan synthase is a monomer
Biochim. Biophys. Acta
1770
286-290
2007
Pasteurella multocida
Tracy, B.S.; Avci, F.Y.; Linhardt, R.J.; DeAngelis, P.L.
Acceptor specificity of the Pasteurella hyaluronan and chondroitin synthases and production of chimeric glycosaminoglycans
J. Biol. Chem.
282
337-344
2007
Pasteurella multocida
Mao, Z.; Chen, R.R.
Recombinant synthesis of hyaluronan by Agrobacterium sp
Biotechnol. Prog.
23
1038-1042
2007
Pasteurella multocida (Q7BLV3), Pasteurella multocida
Kooy, F.; Beeftink, H.; Eppink, M.; Tramper, J.; Eggink, G.; Boeriu, C.
Kinetic and structural analysis of two transferase domains in Pasteurella multocida hyaluronan synthase
J. Mol. Catal. B
102
138-145
2014
Pasteurella multocida (Q7BLV3)
-
Vigetti, D.; Viola, M.; Karousou, E.; De Luca, G.; Passi, A.
Metabolic control of hyaluronan synthases
Matrix Biol.
35
8-13
2014
Mammalia, Streptococcus sp., Pasteurella multocida (Q7BLV3)
Eisele, A.; Zaun, H.; Kuballa, J.; Elling, L.
In vitro one-pot enzymatic synthesis of hyaluronic acid from sucrose and N-acetylglucosamine optimization of the enzyme module system and nucleotide sugar regeneration
ChemCatChem
10
2969-2981
2018
Pasteurella multocida (Q7BLV3)
-
EXTERNAL LINKS (specific for EC-Number 2.4.1.212)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
