Information on EC 4.2.2.5 - chondroitin AC lyase

New: Word Map on EC 4.2.2.5
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
4.2.2.5
-
RECOMMENDED NAME
GeneOntology No.
chondroitin AC lyase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Eliminative degradation of polysaccharides containing 1,4-beta-D-hexosaminyl and 1,3-beta-D-glucuronosyl linkages to disaccharides containing 4-deoxy-beta-D-gluc-4-enuronosyl groups
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
elimination
-
-
-
-
additional information
-
no beta-elimination
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
chondroitin sulfate degradation I (bacterial)
-
-
SYSTEMATIC NAME
IUBMB Comments
chondroitin AC lyase
Acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate. In general, chondroitin sulfate (CS) and dermatan sulfate (DS) chains comprise a linkage region, a chain cap and a repeat region. The repeat region of CS is a repeating disaccharide of glucuronic acid (GlcA) and N-acetylgalactosamine (GalNAc) [-4)GlcA(beta1-3)GalNAc(beta1-]n, which may be O-sulfated on the C-4 and/or C-6 of GalNAc and C-2 of GlcA. GlcA residues of CS may be epimerized to iduronic acid (IdoA) forming the repeating disaccharide [-4)IdoA(alpha1-3)GalNAc(beta1-]n of DS. Both the concentrations and locations of sulfate-ester substituents vary with glucosaminoglycan source [4].
CAS REGISTRY NUMBER
COMMENTARY hide
9047-57-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
Arthrobacter aurescens
strain HJ-15, human intestinal anaerobic bacterium
-
-
Manually annotated by BRENDA team
strain G4
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
-
chondroitin AC I lyase, an enzyme belonging to the class of lyases, which due to its endolytic activity cleaves the glycosidic bonds between GalNAc and D-GlcA for depolymerization of chondroitin sulfate and dermatan sulfate
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiruronic acid
?
show the reaction diagram
-
-
-
-
?
benzyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
benzyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
benzyl 4-O-(2'-nitrophenyl)-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
synthetic chromogenic substrate
-
?
benzyl 4-O-(3',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
synthetic chromogenic substrate
-
?
benzyl 4-O-(4'-chloro-2'-nitrophenyl)-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
synthetic chromogenic substrate
-
?
benzyl S-(2-acetamido-2-deoxy-beta-D-galactopyranosyl)-(1-4)-4-deoxy-4-thio-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
very slow cleavage, 9% product after 24 h
-
-
?
chondroitin
?
show the reaction diagram
chondroitin 4-sulfate
2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose
show the reaction diagram
chondroitin 4-sulfate
?
show the reaction diagram
chondroitin 6-sulfate
?
show the reaction diagram
chondroitin sulfate A
?
show the reaction diagram
chondroitin sulfate A
disaccharides
show the reaction diagram
-
i.e. chondroitin 4-sulfate
-
?
chondroitin sulfate C
?
show the reaction diagram
chondroitin sulfate C
disaccharides
show the reaction diagram
-
i.e. chondroitin 6-sulfate
product analysis
?
chondroitin sulfate/dermatan sulfate chain
?
show the reaction diagram
-
chondroitin AC I lyase cleaves the linkage between GalNAc and D-GlcA, with the formation of a 4,5 double bond at HexA and a water molecule elimination
-
-
?
chondroitin/dermatan sulfate chain
?
show the reaction diagram
-
chon-AC-lyase cleaves the linkage between GalNAc and D-GlcA
-
-
?
dermatan sulfate
?
show the reaction diagram
GlcNAc-D-glucuronic acid-GlcNAc-D-glucuronic acid
DELTA-4,5-uronic acid-GlcNAc + N-acetyl-D-glucosamine + DELTA-4,5-glucuronic acid
show the reaction diagram
Arthrobacter aurescens
-
-
-
?
heparan sulfate
?
show the reaction diagram
-
wild-type enzyme and mutants R292A, R288A, and Y234F
-
?
hyaluronan
?
show the reaction diagram
hyaluronan methyl ester
?
show the reaction diagram
Arthrobacter aurescens
-
-
-
-
?
hyaluronate
?
show the reaction diagram
hyaluronic acid
?
show the reaction diagram
methyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
synthetic chromogenic substrate
-
?
phenyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
phenyl 4-methylumbelliferyl-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
phenyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
-
-
-
?
proteoglycan
protein-keratan sulfate
show the reaction diagram
-
-
the protein-keratan sulfate core isolated after limit digestion of proteoglycan contains about 44% protein, 38% keratan sulfate, and 18% of the enzymatically modified oligosaccharide attachment region between the chondroitin sulfate chains and protein core: gluco-4-enepyranosyluronic acid-(galactosyl)2-xylose
?
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
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
Arthrobacter aurescens
-
activates more effectively than univalent cations
K+
Arthrobacter aurescens
-
activates
Mn2+
-
1 mM, activates
Na+
Arthrobacter aurescens
-
activates
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(13Z)-docos-13-enoic acid
arachidic acid
behenic acid
benzyl 4-deoxy-5-fluoro-beta-D-galactopyranosiduronic acid
-
competitive inhibition
benzyl S-(2-acetamido-2-deoxy-beta-D-galactopyranosyl)-(1-4)-4-deoxy-4-thio-beta-D-glucopyranosiduronic acid
-
competitive versus phenyl 4-O -(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
capric acid
dermatan sulfate
Arthrobacter aurescens
-
-
eicosadienoic acid
eicosanoic acid
eicosapentaenoic acid
eicosatetraenoic acid
eicosatrienoic acid
elaidic acid
lauric acid
linoleic acid
linolenic acid
methyl (9Z)-octadecenoate
myristic acid
myristoleic acid
nervonic acid
oleic acid
p-Chloromercuriphenyl sulfonic acid
-
strong inhibition
palmitic acid
palmitoleic acid
Pb2+
Arthrobacter aurescens
-
-
petroselinic acid
phenyl 4-deoxy-4,4-difluoro-beta-D-xylo-hexopyranosiduronic acid
-
competitive inhibition
ricinoleic acid
Sn2+
Arthrobacter aurescens
-
-
stearic acid
vaccenic acid
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
chondroitin 4-sulfate
-
2.5fold induction of enzyme expression
additional information
-
not affected by Mg2+, Mn2+, Ca2+, Co2+, and EDTA
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6.9
4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiruronic acid
-
pH 6.8, 30C, wild-type enzyme
28
benzyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
7
benzyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
16.9
benzyl 4-O-(2'-nitrophenyl)-beta-D-glucopyranosiduronic acid
-
pH 6.8
9
benzyl 4-O-(3',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
-
pH 6.8
12.2
benzyl 4-O-(4'-chloro-2'-nitrophenyl)-beta-D-glucopyranosiduronic acid
-
pH 6.8
0.0322 - 0.273
Chondroitin
0.00544 - 0.8
chondroitin 4-sulfate
0.00528 - 0.8
chondroitin 6-sulfate
0.38 - 0.42
chondroitin sulfate A
0.008 - 0.167
chondroitin sulfate C
0.1755
hyaluronan
Arthrobacter aurescens
-
-
0.00119
hyaluronan methyl ester
Arthrobacter aurescens
-
-
0.217 - 0.239
hyaluronate
0.00286 - 0.259
hyaluronic acid
114
phenyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
0.9 - 1
phenyl 4-methylumbelliferyl-beta-D-glucopyranosiduronic acid
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.041
4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiruronic acid
Pedobacter heparinus
-
pH 6.8, 30C, wild-type enzyme
0.16
benzyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
0.019
benzyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
Pedobacter heparinus
-
pH 6.8
0.027
benzyl 4-O-(2'-nitrophenyl)-beta-D-glucopyranosiduronic acid
Pedobacter heparinus
-
pH 6.8
0.019
benzyl 4-O-(3',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
Pedobacter heparinus
-
pH 6.8
0.035
benzyl 4-O-(4'-chloro-2'-nitrophenyl)-beta-D-glucopyranosiduronic acid
Pedobacter heparinus
-
pH 6.8
0.005
benzyl S-(2-acetamido-2-deoxy-beta-D-galactopyranosyl)-(1-4)-4-deoxy-4-thio-beta-D-glucopyranosiduronic acid
Pedobacter heparinus
-
-
840 - 868
Chondroitin
234 - 403
chondroitin 4-sulfate
345 - 855
chondroitin 6-sulfate
570 - 1200
hyaluronate
2.3
phenyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
0.0015 - 0.0016
phenyl 4-methylumbelliferyl-beta-D-glucopyranosiduronic acid
additional information
additional information
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3
benzyl 4-deoxy-5-fluoro-beta-D-galactopyranosiduronic acid
-
-
45
benzyl S-(2-acetamido-2-deoxy-beta-D-galactopyranosyl)-(1-4)-4-deoxy-4-thio-beta-D-glucopyranosiduronic acid
-
-
0.0211
eicosatrienoic acid
-
pH 6.0, 37C
0.0047
nervonic acid
Arthrobacter aurescens
-
pH 6.0, 37C
24
phenyl 4-deoxy-4,4-difluoro-beta-D-xylo-hexopyranosiduronic acid
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.012
-
purified mutant R292A, substrate dermatan sulfate
0.028
-
purified mutant Y234F, substrate dermatan sulfate
0.031
-
purified mutant H225A, substrate chondroitin 4-sulfate
0.038
-
purified mutant R288A, substrate heparan sulfate
0.077
-
purified mutant R292A, substrate heparan sulfate
0.12
-
purified mutant Y234F, substrate heparan sulfate
0.18
-
purified transconjugated wild-type, substrate heparan sulfate
0.2
-
purified transconjugated wild-type, substrate dermatan sulfate
0.23
-
purified mutant Y234F, substrate chondroitin 6-sulfate
0.3
-
purified mutant Y234F, substrate hyaluronic acid
0.45
-
purified mutant R288A, substrate hyaluronic acid
0.53
-
purified mutant Y234F, substrate chondroitin 4-sulfate
0.55
-
purified mutant R288A, substrate chondroitin 6-sulfate
0.94
-
purified mutant R288A, substrate chondroitin 4-sulfate
3.63
-
purified mutant R292A, substrate hyaluronic acid
9.54
-
purified mutant R292A, substrate chondroitin 6-sulfate
21.2
-
purified mutant R292A, substrate chondroitin 4-sulfate
57.03
-
purified enzyme
62.5
-
purified transconjugated wild-type, substrate chondroitin 6-sulfate
68
-
chondroitin 4-sulfate
94.5
-
purified transconjugated wild-type, substrate hyaluronic acid
99.7
-
-
111
-
chondroitin 6-sulfate
123.9
-
purified transconjugated wild-type, substrate chondroitin 4-sulfate
541
-
purified recombinant enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.3
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
Arthrobacter aurescens
-
pH 5: about 25% of maximal activity, pH 8: about 35% of maximal activity
5 - 9.5
-
pH 5.0: about 30% of maximal activity, pH 9.5: about 40% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
Arthrobacter aurescens
-
-
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 25
-
activity is significantly higher at 25C than at 20C
25 - 55
-
25C: about 85% of maximal activity, 55C: about 45% of maximal activity
additional information
Arthrobacter aurescens
-
-
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.19
-
isoelectric focusing, pH-range 3.5-10
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Arthrobacter aurescens
Arthrobacter aurescens
Arthrobacter aurescens
Arthrobacter aurescens
Arthrobacter aurescens
Arthrobacter aurescens
Pedobacter heparinus (strain ATCC 13125 / DSM 2366 / NCIB 9290)
Pedobacter heparinus (strain ATCC 13125 / DSM 2366 / NCIB 9290)
Pedobacter heparinus (strain ATCC 13125 / DSM 2366 / NCIB 9290)
Pedobacter heparinus (strain ATCC 13125 / DSM 2366 / NCIB 9290)
Pedobacter heparinus (strain ATCC 13125 / DSM 2366 / NCIB 9290)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
42600
-
gel filtration
71000
-
gel filtration
76000
Arthrobacter aurescens
-
gel filtration
78030
-
mass spectrometry
79500 - 79840
Arthrobacter aurescens
-
mass spectrometry
170000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
Arthrobacter aurescens
-
1 * 75000-80000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
-
the enzyme contains a N-terminal signal peptide which is proteolytically cleaved to produce an active enzyme
side-chain modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
-
Arthrobacter aurescens
-
native enzyme alone and in complex with substrates chondroitin 4-sulfate tetrasaccharide, and hyaluronan tetrasaccharide, hanging drop vapour diffusion method, 0.002 ml protein solution: 10 mg/ml protein, + 0.002 ml reservoir solution: 23% w/v PEG 8000, 0.1 M sodium phosphate, pH 6.4, 0.4 M ammonium acetate, 10% v/v glycerol, suspended over 1 ml reservoir solution, a few days, larger crystals by macroseeding, X-ray diffraction structure determination and analysis of the enzyme-substrate complexes at 1.25-1.9 A high resolution
Arthrobacter aurescens
-
complexes of the wild-type enzyme with dermatan sulfate hexasaccharide, tetrasaccharide, and hyaluronic acid tetrasaccharide, and mutant Y234F enzyme in complex with chondroitin sulfate tetrasaccharide, hanging drop vapour diffusion method, equal volumes, 0.005 ml, of protein and reservoir solution are suspended over 1 ml reservoir solution containing 15% w/v PEG 3350, 0.4 M sodium acetate, 0.1 M HEPES, pH 7.5, at room temperature of about 20C, 1 day, X-ray diffraction structure determination and analysis at 2.0-2.3 A resolution
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 9
-
stable
34098
6 - 7
Arthrobacter aurescens
-
40C, 2 h, no loss of activity
34092
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
-
6 h, 50% loss of activity
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme
-
recombinant His-tagged active enzyme from Escherichia coli, 66fold
-
recombinant mutant from mutant Flavobacterium heparinum strain
-
to homogeneity
-
using His bind column chromatography
wild-type enzyme, and recombinant mutant enzymes from Flavobacterium heparinum mutant strains
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning and construction of mutants in Escherichia coli and expression in Flavobacterium heparinum, the Flavobacterium heparinum strains are mutated by conjugation
-
expressed in Escherichia coli as a His-tagged fusion protein
gene cAC, DNA sequence determination and analysis, functional expression without the N-terminal signal peptide in Escherichia coli as His-tagged protein, 2 different expression systems
-
gene cslA, DNA sequence determination and analysis, chromosomal localization, expression in Escherichia coli
-
mutant R292A, expression in Escherichia coli and conjugation to Flavobacterium heparinum to create a mutant strain
-
the gene cslA is expressed in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H225A
-
site-directed mutagenesis, active site residue, inactive mutant
R288A
-
site-directed mutagenesis, active site residue, nearly inactive mutant
additional information
a vector is engineered containing the gene for Flavobacterium heparinum chondroitinase AC for expression in adult bone marrow-derived cells which are then transplanted into an injury site in the CNS. Expression and secretion of active chondroitinase AC is observed in vitro using transfected Chinese hamster ovarian and gliosarcoma cells and in vivo by immunohistochemistry analysis which show degraded chondroitin sulfate coinciding with the location of transfected bone marrow-derived cells. Immunolabelling of the axonal growth-associated protein GAP-43 is observed in vivo and coincides with the location of degraded chondroitin sulfate
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
useful tool for the identification and structural characterization of chondriotin and dermatan sulfates
food industry
-
in Flavobacterium columnare strains C, E, G and H isolated from disease outbreaks, chondroitinase activity is significantly higher in the virulent, rhizoid variants than in the rough variants of the same strain. Temperature significantly increases the adhesion of rhizoid variants up to 20C. Both rhizoid colony morphology and high chondroitinase activity seem to be needed for virulence and temperature may promote the adhesion of the virulent variants to surfaces at fish farms
medicine
bone marrow-derived mononuclear cells, transfected with our construct and transplanted into CNS, could be a potential tool for studying an alternative chondroitinase AC delivery method