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.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
strain NEM316
UniProt
Manually annotated by BRENDA team
strain M1 GAS SF370
UniProt
Manually annotated by BRENDA team
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, 37C
0.16
2,4-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37C
0.59
2,5-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37C
4.5
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-4-O-sulfo-D-galactopyranose
-
pH 6.6, 37C
3.7
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-6-O-sulfo-D-galactopyranose
-
pH 6.6, 37C
3
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-D-galactopyranose
-
pH 6.6, 37C
1.2
2-deoxy-4-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-2-(sulfoamino)-D-glucopyranose
-
pH 6.6, 37C
0.88
3-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37C
0.84
4-chlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37C
0.26
4-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37C
0.88
4-tert-butylphenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37C
9
benzyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
-
pH 6.6, 37C
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, 37C
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
Clostridium perfringens
-
pH 6.6, 37C
0.14
2,4-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
0.64
2,5-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
0.09
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-4-O-sulfo-D-galactopyranose
Clostridium perfringens
-
pH 6.6, 37C
112
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-6-O-sulfo-D-galactopyranose
Clostridium perfringens
-
pH 6.6, 37C
8.5
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-D-galactopyranose
Clostridium perfringens
-
pH 6.6, 37C
14
2-deoxy-4-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-2-(sulfoamino)-D-glucopyranose
Clostridium perfringens
-
pH 6.6, 37C
14.7
3-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
3.9
4-chlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
2.05
4-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
3.27
4-tert-butylphenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
18.6
benzyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
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
Clostridium perfringens
-
pH 6.6, 37C
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
Clostridium perfringens
-
pH 6.6, 37C
199532
0.9
2,4-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
199530
1.1
2,5-dinitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
199531
0.02
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-4-O-sulfo-D-galactopyranose
Clostridium perfringens
-
pH 6.6, 37C
199543
30
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-6-O-sulfo-D-galactopyranose
Clostridium perfringens
-
pH 6.6, 37C
199539
2.8
2-(acetylamino)-2-deoxy-3-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-D-galactopyranose
Clostridium perfringens
-
pH 6.6, 37C
199541
12
2-deoxy-4-O-(4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl)-2-(sulfoamino)-D-glucopyranose
Clostridium perfringens
-
pH 6.6, 37C
199540
16
3-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
199534
4.6
4-chlorophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
199535
0.000044
4-deoxy-alpha-L-threo-hex-4-enopyranuronosyl fluoride
Clostridium perfringens
-
pH 6.6, 37C
199529
7.9
4-nitrophenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
199533
3.7
4-tert-butylphenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
199537
2.1
benzyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
199538
2.12 - 10.7
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc
4181
1.63 - 130
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S
2921
0.086 - 2
DELTAGlcUA-beta-(1->4)-GlcN2S
202179
1.3
phenyl 4-deoxy-alpha-L-threo-hex-4-enopyranosiduronic acid
Clostridium perfringens
-
pH 6.6, 37C
199536
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, 37C
3
(3R,4S,5R)-3,4,5-trihydroxycyclohex-1-ene-1-carboxylic acid
Clostridium perfringens
-
pH 6.6, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.7
purified recombinant wild-type enzyme, pH 7.5, 30C, substrate DELTAGlcUA-beta-(1->3)-GalNAc6S
6.1
-
purified recombinant wild-type enzyme, pH 7.5, 30C, substrate DELTAGlcUA-beta-(1,3)-GalNAc6S
12
purified recombinant wild-type enzyme, pH 7.5, 30C, substrate DELTAGlcUA-beta-(1->4)-GlcNAc
15
purified recombinant wild-type enzyme, pH 7.5, 30C, substrate DELTAGlcUA-beta-(1->4)-GlcNAc
17
-
purified recombinant wild-type enzyme, pH 7.5, 30C, substrate DELTAGlcUA-beta-(1->3)-GalNAc6S
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Streptococcus agalactiae serotype III (strain NEM316)
Streptococcus agalactiae serotype III (strain NEM316)
Streptococcus agalactiae serotype III (strain NEM316)
Streptococcus agalactiae serotype III (strain NEM316)
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, 20C, 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
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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
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no hydrolysis of beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S
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D175N
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no hydrolysis of beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S
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R236A
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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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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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