Information on EC 3.2.1.180 - unsaturated chondroitin disaccharide hydrolase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.2.1.180
-
RECOMMENDED NAME
GeneOntology No.
unsaturated chondroitin disaccharide hydrolase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
beta-D-4-deoxy-DELTA4-GlcAp-(1->3)-beta-D-GalNAc6S + H2O = 5-dehydro-4-deoxy-D-glucuronate + N-acetyl-beta-D-galactosamine-6-O-sulfate
show the reaction diagram
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
chondroitin sulfate degradation I (bacterial)
-
-
hyaluronan degradation
-
-
SYSTEMATIC NAME
IUBMB Comments
beta-D-4-deoxy-DELTA4-GlcAp-(1->3)-beta-D-GalNAc6S hydrolase
The enzyme releases 4-deoxy-4,5-didehydro D-glucuronic acid or 4-deoxy-4,5-didehydro L-iduronic acid from chondroitin disaccharides, hyaluronan disaccharides and heparin disaccharides and cleaves both glycosidic (1->3) and (1->4) bonds. It prefers the sulfated disaccharides to the unsulfated disaccharides.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
-
the enzyme is involved in the metabolic pathway for degradation of glycosaminoglycans
physiological function
-
unsaturated glucuronyl hydrolases acts on the unsaturated uronic-acid-containing oligosaccharides to remove the unusual non-reducing terminal monomer units
additional information
-
a hydration-initiated mechanism with a transition state with positive charge at carbon 5 and the endocyclic oxygen is proposed, transition-state structure for the first irreversible step of this mechanism, overview
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,4,6-trichlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid + H2O
?
show the reaction diagram
-
-
-
-
?
2,4-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid + H2O
?
show the reaction diagram
-
-
-
-
?
2,5-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid + H2O
?
show the reaction diagram
-
-
-
-
?
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-4-O-sulfo-D-galactopyranose + H2O
?
show the reaction diagram
-
-
-
-
?
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-6-O-sulfo-D-galactopyranose + H2O
?
show the reaction diagram
-
-
-
-
?
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-D-galactopyranose + H2O
?
show the reaction diagram
-
-
-
-
?
2-acetamido-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-6-O-sulfo-D-galactopyranose + H2O
2-acetamido-2-deoxy-6-O-sulfo-D-galactopyranose + 4-deoxy-alpha-L-threo-hex-4-enopyranose
show the reaction diagram
-
high activity, a substrate with sulfation of the primary alcohol on carbon 6 of its N-acetylgalactosamine moiety
-
-
?
2-deoxy-4-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-2-(sulfoamino)-D-glucopyranose + H2O
?
show the reaction diagram
-
-
-
-
?
3-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid + H2O
?
show the reaction diagram
-
-
-
-
?
4-chlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid + H2O
?
show the reaction diagram
-
-
-
-
?
4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl fluoride + H2O
?
show the reaction diagram
-
an unsaturated glucuronyl fluoride giving very low activity
-
-
?
4-deoxy-beta-L-threo-hex-4-enopyranuronosyl fluoride + H2O
?
show the reaction diagram
-
an unsaturated glucuronyl fluoride giving very low activity
-
-
?
4-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid + H2O
?
show the reaction diagram
-
-
-
-
?
4-tert-butylphenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid + H2O
?
show the reaction diagram
-
-
-
-
?
alpha-D-4-deoxy-DELTA4,5-IdoA-(1->4)-beta-D-GlcNAc + H2O
alpha-D-4-deoxy-DELTA4,5-IdoA + beta-D-N-acetylglucosamine
show the reaction diagram
-
-
-
-
?
benzyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid + H2O
?
show the reaction diagram
-
-
-
-
?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc + H2O
4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine
show the reaction diagram
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S + H2O
4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine-6-O-sulfate
show the reaction diagram
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GlcNAc + H2O
4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylglucosamine
show the reaction diagram
DELTA4GlcUA2S-beta-(1->4)-GlcN2S + H2O
?
show the reaction diagram
-
-
-
-
?
DELTAGlcUA-beta-(1,3)-GalNAc + H2O
?
show the reaction diagram
-
-
-
-
?
DELTAGlcUA-beta-(1,3)-GalNAc4S + H2O
?
show the reaction diagram
-
-
-
-
?
DELTAGlcUA-beta-(1,3)-GalNAc6S + H2O
?
show the reaction diagram
-
best substrate
-
-
?
DELTAGlcUA-beta-(1,4)-GlcN2S + H2O
?
show the reaction diagram
-
-
-
-
?
DELTAGlcUA-beta-(1,4)-GlcN2S6S + H2O
?
show the reaction diagram
-
-
-
-
?
DELTAGlcUA-beta-(1,4)-GlcN6S + H2O
?
show the reaction diagram
-
-
-
-
?
DELTAGlcUA-beta-(1,4)-GlcNAc + H2O
?
show the reaction diagram
-
-
-
-
?
DELTAGlcUA-beta-(1,4)-GlcNAc6S + H2O
?
show the reaction diagram
-
-
-
-
?
DELTAGlcUA-beta-(1->3)-GalNAc + H2O
?
show the reaction diagram
DELTAGlcUA-beta-(1->3)-GalNAc4S + H2O
?
show the reaction diagram
DELTAGlcUA-beta-(1->3)-GalNAc6S + H2O
?
show the reaction diagram
DELTAGlcUA-beta-(1->4)-GlcN2S + H2O
?
show the reaction diagram
DELTAGlcUA-beta-(1->4)-GlcN2S6S + H2O
?
show the reaction diagram
DELTAGlcUA-beta-(1->4)-GlcN6S + H2O
?
show the reaction diagram
DELTAGlcUA-beta-(1->4)-GlcNAc + H2O
?
show the reaction diagram
DELTAGlcUA-beta-(1->4)-GlcNAc6S + H2O
?
show the reaction diagram
DELTAGlcUA2S-beta-(1,4)-GlcN2S + H2O
?
show the reaction diagram
-
-
-
-
?
DELTAGlcUA2S-beta-(1->4)-GlcN2S + H2O
?
show the reaction diagram
-
-
-
-
?
phenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2R)-pyrrolidinium-2-carboxylate
-
-
(2R,3R,4S,5S,6R)-3,4,5,6-tetrahydroxypiperidine-2-carboxylic acid
-
competitive
(2S)-pyrrolidinium-2-carboxylate
-
-
(3R,4S,5R)-3,4,5-trihydroxycyclohex-1-ene-1-carboxylic acid
-
competitive
2-mercaptoethanol
1 mM, 20% inhibition
AlCl3
-
1 mM, 65% inhibition
CoCl2
-
1 mM, 55% inhibition
D-galactose
5 mM, 21% inhibition
D-galacturonic acid
D-GlcA
5 mM, 97% inhibition
D-glucose
5 mM, 40% inhibition
D-glucuronic acid
D-mannose
5 mM, 22% inhibition
D-xylose
5 mM, 33% inhibition
KCl
-
1 mM, 22% inhibition
L-fucose
5 mM, 50% inhibition
L-rhamnose
5 mM, 35% inhibition
LiCl
-
1 mM, 24% inhibition
MnCl2
-
1 mM, 27% inhibition
N-acetylglucosamine
-
5 mM, 17% inhibition
NaCl
-
1 mM, 22% inhibition
additional information
-
several analogues designed to take advantage ofstrong enzyme binding to the reaction's transition state show little to no inhibition. No inhibition by 17a and 17b
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.58
2,4,6-trichlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.16
2,4-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.59
2,5-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
4.5
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-4-O-sulfo-D-galactopyranose
-
pH 6.6, 37°C
3.7
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-6-O-sulfo-D-galactopyranose
-
pH 6.6, 37°C
3
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-D-galactopyranose
-
pH 6.6, 37°C
1.2
2-deoxy-4-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-2-(sulfoamino)-D-glucopyranose
-
pH 6.6, 37°C
0.88
3-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.84
4-chlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.26
4-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.88
4-tert-butylphenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
9
benzyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.371 - 1.4
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc
0.1 - 2.36
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S
0.86 - 5.2
DELTAGlcUA-beta-(1->4)-GlcN2S
0.029 - 1.4
DELTAGlcUA-beta-(1->4)-GlcNAc
3.2
phenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
additional information
additional information
-
Michaelis-Menten kinetic parameters for the cleavage of a variety of unsaturated disaccharides of glycosaminoglycan origin, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.312
2,4,6-trichlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.14
2,4-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.64
2,5-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.09
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-4-O-sulfo-D-galactopyranose
-
pH 6.6, 37°C
112
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-6-O-sulfo-D-galactopyranose
-
pH 6.6, 37°C
8.5
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-D-galactopyranose
-
pH 6.6, 37°C
14
2-deoxy-4-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-2-(sulfoamino)-D-glucopyranose
-
pH 6.6, 37°C
14.7
3-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
3.9
4-chlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
2.05
4-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
3.27
4-tert-butylphenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
18.6
benzyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
1.69 - 10.8
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc
1.3 - 41.6
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S
0.075 - 6
DELTAGlcUA-beta-(1->4)-GlcN2S
16 - 22
DELTAGlcUA-beta-(1->4)-GlcNAc
4.3
phenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.55
2,4,6-trichlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.9
2,4-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
1.1
2,5-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.02
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-4-O-sulfo-D-galactopyranose
-
pH 6.6, 37°C
30
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-6-O-sulfo-D-galactopyranose
-
pH 6.6, 37°C
2.8
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-D-galactopyranose
-
pH 6.6, 37°C
12
2-deoxy-4-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-2-(sulfoamino)-D-glucopyranose
-
pH 6.6, 37°C
16
3-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
4.6
4-chlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
0.000044
4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl fluoride
-
pH 6.6, 37°C
7.9
4-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
3.7
4-tert-butylphenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
2.1
benzyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
2.12 - 10.7
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc
1.63 - 130
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S
0.086 - 2
DELTAGlcUA-beta-(1->4)-GlcN2S
1.3
phenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37°C
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.2
(2R,3R,4S,5S,6R)-3,4,5,6-tetrahydroxypiperidine-2-carboxylic acid
Clostridium perfringens;
-
pH 6.6, 37°C
3
(3R,4S,5R)-3,4,5-trihydroxycyclohex-1-ene-1-carboxylic acid
Clostridium perfringens;
-
pH 6.6, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.7
purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1->3)-GalNAc6S
6.1
-
purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1,3)-GalNAc6S
12
purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1->4)-GlcNAc
15
purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1->4)-GlcNAc
17
-
purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1->3)-GalNAc6S
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PDB
SCOP
CATH
UNIPROT
ORGANISM
Streptococcus agalactiae serotype III (strain NEM316);
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46079
1 * 46079, calculated from sequence
46100
1 * 46100, calculated from sequence
46587
-
1 * 46587, calculated from sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant wild-type enzyme, sitting drop vapor diffusion method, mixing of 0..001 ml of 10 mg/ml protein in n 20 mM Tris-HCl, pH 7.5, 1 mM dithiothreitol, and 1 mM EDTA, with 0.001 ml of reservoir solution containing 25% PEG 6000, 0.1 M HEPES, pH 7.5, and 0.1 M lithium chloride, 20°C, 2 months, X-ray diffraction structure determination and analysis at 1.3 A resolution, molecular replacement
hanging-drop vapor-diffusion method, crystal structure at 1.75 A resolution
-
sitting-drop vapor-diffusion method
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme from Escherichia coli
recombinant tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by affinity chromatography to near homogeneity
recombinant wild-type and mutant enzymes from Escherichia coli
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
gene Phep_2238, DNA and amino acid sequence determination and analysis, sequence comparisons, expression in Escherichia coli strain BL21(DE3); gene Phep_2649, DNA and amino acid sequence determination and analysis, sequence comparisons, expression in Escherichia coli strain BL21(DE3); gene Phep_2830, DNA and amino acid sequence determination and analysis, sequence comparisons, expression of tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
sequence comparisons
-
sequence comparisons, recombinant expression in Escherichia coli
sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
constitutively expressed, expression level increases in presence of glycosaminoglycan
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D174N
site-directed mutagenesis
D182N
site-directed mutagenesis
F164A
site-directed mutagenesis
G362Y
site-directed mutagenesis
R57A
site-directed mutagenesis
D115N
-
no hydrolysis of beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S
D175N
-
no hydrolysis of beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S
K370A
-
site-directed mutagenesis
K370I
-
kcat/KM for beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S is 28% of the wild-type value, kcat/Km for beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc is 5fold higher than wild-type value
R236A
-
effectively degrades beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S. Almost inactive on beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc4S, although the substrate is degraded by the wild-type enzyme
R236H
-
effectively degrades beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S. Almost inactive on beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc4S, although the substrate is degraded by the wild-type enzyme
S365A
-
site-directed mutagenesis
S365G
-
site-directed mutagenesis
S365H
-
kcat/KM for beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S is 1.6% of the wild-type value, kcat/Km for beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc is similar to wild-type value
S368A
-
site-directed mutagenesis
T235A
-
kcat/KM for beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S is 2% of the wild-type value
D115N
-
no hydrolysis of beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S
-
D175N
-
no hydrolysis of beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S
-
R236A
-
effectively degrades beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S. Almost inactive on beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc4S, although the substrate is degraded by the wild-type enzyme
-
R236H
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effectively degrades beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S. Almost inactive on beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc4S, although the substrate is degraded by the wild-type enzyme
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