Information on EC 4.2.2.5 - chondroitin AC lyase

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

EC NUMBER
COMMENTARY
4.2.2.5
-
RECOMMENDED NAME
GeneOntology No.
chondroitin AC lyase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
-
-
-
-
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
stepwise attack
-
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
random attack
-
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
acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate, random endolytic elimination mechanism
-
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
acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate, mechanism, active site structure, His225, Tyr234, Arg288, and Glu371 form a catalytic tetrad
-
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
acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate, reaction mechanism
-
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
acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate
-
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
acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate, stepwise reaction mechanism
-
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
acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate
-
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
acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate
-
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
acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate, lytic mechanism with Tyr242 acting as a general base that abstracts the proton from the C5 position of glucuronic acid while Asn183 and His233 neutralize the charge on the glucuronate acidic group, substrate binding site
-
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
acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate
Bacteroides stercoris HJ-15
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
elimination
-
-
-
-
additional information
-
no beta-elimination
PATHWAY
KEGG Link
MetaCyc Link
chondroitin sulfate and dermatan 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].
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A-Chase
-
-
-
-
AC I lyase
-
-
ChnAC
Q59288
-
chon-AC-lyase
-
-
ChonAC
Flavobacterium columnare G4
-
-
-
chondroitin AC eliminase
-
-
-
-
chondroitin AC I lyase
-
-
chondroitin AC I lyase
-
-
Chondroitin AC lyase
-
-
-
-
Chondroitin AC lyase
-
-
Chondroitin AC lyase
Flavobacterium columnare G4
-
-
-
Chondroitin AC lyase
-
-
chondroitin ACII lyase
-
-
chondroitin lyase AC
-
-
chondroitin sulfate lyase
-
-
-
-
chondroitinase
-
-
-
-
chondroitinase AC
-
-
-
-
chondroitinase AC
A9Y5J0
-
chondroitinase AC
-
-
chondroitinase AC
Q59288
-
chondroitinase ACI
-
-
chondroitinase ACII
-
-
ChSase-AC
-
-
-
-
EC 4.2.99.6
-
-
formerly
-
F-Chase
-
-
-
-
lyase, chondroitin AC
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9047-57-8
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
multiple forms: chondroitinase I, chondroitinase II and chondroitinase III
-
-
Manually annotated by BRENDA team
strain HJ-15, human intestinal anaerobic bacterium
-
-
Manually annotated by BRENDA team
Bacteroides stercoris HJ-15
strain HJ-15, human intestinal anaerobic bacterium
-
-
Manually annotated by BRENDA team
Flavobacterium columnare G4
strain G4
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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 ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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
-
synthetic chromogenic substrate
-
?
benzyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
synthetic chromogenic substrate
-
?
benzyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
synthetic chromogenic substrate
-
?
benzyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
synthetic chromogenic substrate
-
?
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
-
best substrate
-
?
chondroitin
?
show the reaction diagram
-
degradation is faster than with chondroitin 4-sulfate or chondroitin 6-sulfate
-
-
?
chondroitin
?
show the reaction diagram
-
degradation is faster than with chondroitin 4-sulfate or chondroitin 6-sulfate
-
-
?
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
2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose
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
2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose
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
2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose
show the reaction diagram
-
i.e. chondroitin sulfate A
-
-
-
chondroitin 4-sulfate
?
show the reaction diagram
-
ACII lyase is an exolytic enzyme able to cut the polysaccharide chains starting from the nonreducing end
-
-
?
chondroitin 4-sulfate
?
show the reaction diagram
-
chondroitin AC lyase is specific for chondroitin 4-sulfate and has endolytic activity
-
-
?
chondroitin 4-sulfate
?
show the reaction diagram
-
exolytic action pattern
the products afforded by chondroitinase ACII contain primarily unsaturated disaccharide
-
?
chondroitin 4-sulfate
?
show the reaction diagram
-
random endolytic action pattern
the products of chondroitinase AC contain unsaturated oligosaccharides of sizes ranging from disaccharide to decasaccharide
-
?
chondroitin 6-sulfate
?
show the reaction diagram
-
-
-
-
?
chondroitin 6-sulfate
?
show the reaction diagram
-
-
-
-
?
chondroitin 6-sulfate
?
show the reaction diagram
-
-
-
-
?
chondroitin 6-sulfate
?
show the reaction diagram
-
i.e. chondroitin sulfate C
-
-
?
chondroitin 6-sulfate
?
show the reaction diagram
-
the degradation of chondroitin by an anionic E1cb elimination mechanism involves proton abstraction from C5 of glucuronic acid, Tyr234 is the key base catalysts
-
-
?
chondroitin 6-sulfate
?
show the reaction diagram
-
exolytic action pattern
the products afforded by chondroitinase ACII contain primarily unsaturated disaccharide
-
?
chondroitin 6-sulfate
?
show the reaction diagram
-
random endolytic action pattern
the products of chondroitinase AC contain unsaturated oligosaccharides of sizes ranging from disaccharide to decasaccharide
-
?
chondroitin sulfate A
?
show the reaction diagram
-
-
-
-
?
chondroitin sulfate A
?
show the reaction diagram
-
i.e. chondroitin 4-sulfate
-
?
chondroitin sulfate A
?
show the reaction diagram
-
i.e. chondroitin 4-sulfate
-
?
chondroitin sulfate A
?
show the reaction diagram
-
i.e. chondroitin 4-sulfate
-
?
chondroitin sulfate A
?
show the reaction diagram
-
i.e. chondroitin 4-sulfate
-
?
chondroitin sulfate A
?
show the reaction diagram
Q59288
i.e. chondroitin 4-sulfate
-
?
chondroitin sulfate A
?
show the reaction diagram
-
i.e. chondroitin 4-sulfate, best substrate
-
?
chondroitin sulfate A
?
show the reaction diagram
-
i.e. chondroitin 4-sulfate, best substrate, wild-type enzyme and mutants R292A, R288A, H225A and Y234F
-
?
chondroitin sulfate A
?
show the reaction diagram
A9Y5J0, -
relative activity: 100% (recombinant protein), 100% (native protein)
-
-
?
chondroitin sulfate A
?
show the reaction diagram
Bacteroides stercoris HJ-15
-
i.e. chondroitin 4-sulfate, best substrate
-
?
chondroitin sulfate A
disaccharides
show the reaction diagram
-
i.e. chondroitin 4-sulfate
-
?
chondroitin sulfate C
?
show the reaction diagram
-
-
-
-
?
chondroitin sulfate C
?
show the reaction diagram
-
i.e. chondroitin 6-sulfate
-
?
chondroitin sulfate C
?
show the reaction diagram
-
i.e. chondroitin 6-sulfate
-
?
chondroitin sulfate C
?
show the reaction diagram
-
i.e. chondroitin 6-sulfate
-
?
chondroitin sulfate C
?
show the reaction diagram
-
i.e. chondroitin 6-sulfate
-
?
chondroitin sulfate C
?
show the reaction diagram
-
i.e. chondroitin 6-sulfate
-
?
chondroitin sulfate C
?
show the reaction diagram
Q59288
i.e. chondroitin 6-sulfate
-
?
chondroitin sulfate C
?
show the reaction diagram
-
i.e. chondroitin 6-sulfate, low activity
-
?
chondroitin sulfate C
?
show the reaction diagram
-
i.e. chondroitin 6-sulfate, wild-type enzyme and mutants R292A, R288A, and Y234F
-
?
chondroitin sulfate C
?
show the reaction diagram
A9Y5J0, -
relative activity: 42% (recombinant protein), 46% (native protein)
-
-
?
chondroitin sulfate C
?
show the reaction diagram
Bacteroides stercoris HJ-15
-
i.e. chondroitin 6-sulfate, low activity
-
?
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
-
-
-
-
?
dermatan sulfate
?
show the reaction diagram
-
-
-
-
?
dermatan sulfate
?
show the reaction diagram
-
wild-type enzyme and mutants R292A, and Y234F
-
?
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
-
-
-
?
heparan sulfate
?
show the reaction diagram
-
wild-type enzyme and mutants R292A, R288A, and Y234F
-
?
hyaluronan
?
show the reaction diagram
-
-
-
?
hyaluronan
?
show the reaction diagram
-
-
-
-
?
hyaluronan methyl ester
?
show the reaction diagram
-
-
-
-
?
hyaluronate
?
show the reaction diagram
-
-
-
-
-
hyaluronate
?
show the reaction diagram
-
-
-
-
?
hyaluronate
?
show the reaction diagram
-
degradation is faster than with chondroitin 4-sulfate or chondroitin 6-sulfate
-
-
?
hyaluronate
?
show the reaction diagram
-
degradation is faster than with chondroitin 4-sulfate or chondroitin 6-sulfate
-
-
?
hyaluronic acid
?
show the reaction diagram
-
-
-
-
?
hyaluronic acid
?
show the reaction diagram
-
-
-
?
hyaluronic acid
?
show the reaction diagram
-
low activity
-
?
hyaluronic acid
?
show the reaction diagram
-
wild-type enzyme and mutants R292A, R288A, and Y234F
-
?
hyaluronic acid
?
show the reaction diagram
A9Y5J0, -
relative activity: 54% (recombinant protein), 67% (native protein)
-
-
?
hyaluronic acid
?
show the reaction diagram
Bacteroides stercoris HJ-15
-
low activity
-
?
phenyl 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
-
synthetic chromogenic substrate
-
?
phenyl 4-methylumbelliferyl-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
synthetic chromogenic substrate
-
?
phenyl 4-methylumbelliferyl-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
synthetic chromogenic substrate
-
?
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
?
methyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
?
show the reaction diagram
-
synthetic chromogenic substrate
-
?
additional information
?
-
-
does not split endogalactosaminidic bonds neighbouring to reduced disaccharide units
-
-
-
additional information
?
-
-
the enzyme als acts as an exoglycosidase towards hyaluronate
-
-
-
additional information
?
-
-
no activity with dermatan sulfate
-
-
-
additional information
?
-
-
Arg292 is involved in recognition of the N-acetyl and sulfate moieties of galactosamine, but does not participate directly in catalysis
-
?
additional information
?
-
-
developement of an assay using synthetic substrates, which can be monitored by 3 different techniques, UV-Vis spectroscopy, fluorescence spectroccopy, and fluoride ion-selective electrode usage, overview, products of chondroitin sulfate cleavages are disaccharides or oligosaccharides with DELTA4,5-unsaturated uronic acid resdiues at the nonreducing end, methyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid is a poor substrate
-
?
additional information
?
-
-
elimination mechanism, but no syn-elimination, resulting in disaccharides or oligosaccharides with DELTA4,5-unsaturated uronic acid residues at the nonreducing end
-
?
additional information
?
-
-
no activity with dermatan sufate, heparin and heparan sulfate, the latter of porcine or bovine source
-
?
additional information
?
-
-
substrate specificity, no activity with dermatan sulfate
-
?
additional information
?
-
-
the active site forms a tunnel, structurally involved residue Arg292 is responsible for substrate recognition of N-acetyl or O-sulfo moieties in galactosamine residues, it does not participate in catalysis
-
?
additional information
?
-
-
constitutive enzyme
-
-
-
additional information
?
-
-
constitutive enzyme
-
-
-
additional information
?
-
-
no activity with chondroitin sulfate B, heparin or heparan sulfate
-
-
-
additional information
?
-
-
chondroitin AC lyase activity is significantly related to the virulence of the strains at 25C. ChonAC activity is significantly higher in the high virulence group than in the low virulence group at 25C
-
-
-
additional information
?
-
-
digestion of amyloid fibrils with chondroitinase AC hydrolyzes keratan sulfate and heparan sulfate
-
-
-
additional information
?
-
A9Y5J0, -
no activity with dermatan sulfate or heparin as substrate
-
-
-
additional information
?
-
-
chondroitin/dermatan sulfate chain of decorin, with a high complexity due to the large variety of glycoforms, from human skin fibroblasts is released by reductive beta-elimination reaction and digested with chondroitin AC I lyase. Mass spectrometric substrate and product analysis, e.g. tetrasulfated octasaccharide, decasaccharides, a trisulfated, a hexasulfated [4,5-D-GlcAGalNAc(IdoAGalNAc)4], and [4,5-D-GlcAGalNAc(IdoAGalNAc)3], and disaccharides, overview
-
-
-
additional information
?
-
Bacteroides stercoris HJ-15
-
no activity with dermatan sufate, heparin and heparan sulfate, the latter of porcine or bovine source
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
constitutive enzyme
-
-
-
additional information
?
-
-
constitutive enzyme
-
-
-
additional information
?
-
-
chondroitin AC lyase activity is significantly related to the virulence of the strains at 25C. ChonAC activity is significantly higher in the high virulence group than in the low virulence group at 25C
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ba2+
-
activates more effectively than univalent cations
Ba2+
-
1 mM, activates
Ca2+
-
activates more effectively than univalent cations
Ca2+
-
1 mM, activates
Co2+
-
activates more effectively than univalent cations
K+
-
activates
Mg2+
-
activates more effectively than univalent cations
Mg2+
-
1 mM, activates
Mn2+
-
1 mM, activates
Na+
-
activates
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
arachidic acid
-
weak inhibition
behenic acid
-
weak inhibition
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
Cu2+
-
strong inhibition
dermatan sulfate
-
-
Fe2+
-
-
-
myristic acid
-
weak inhibition
myristoleic acid
-
-
myristoleic acid
-
weak inhibition
p-Chloromercuriphenyl sulfonic acid
-
strong inhibition
palmitic acid
-
weak inhibition
phenyl 4-deoxy-4,4-difluoro-beta-D-xylo-hexopyranosiduronic acid
-
competitive inhibition
ricinoleic acid
-
-
ricinoleic acid
-
weak inhibition
stearic acid
-
weak inhibition
Zn2+
-
strong inhibition
methyl (9Z)-octadecenoate
-
weak inhibition
additional information
-
not affected by Mg2+, Mn2+, Ca2+, Co2+, and EDTA
-
additional information
-
degree of inhibition by fatty acids depends on the chain length and the number of unsaturated bonds as well as the stereochemistry, overview
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
pH 6.8, 30C
28
-
benzyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
-
-
7
-
benzyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
-
pH 6.8, 30C
7
-
benzyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
-
pH 6.8
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
-
Chondroitin
-
wild-type enzyme, pH 8.0, 37C
0.119
-
Chondroitin
-
mutant R292A, pH 8.0, 37C
0.239
-
Chondroitin
-
20C
0.273
-
Chondroitin
-
20C
0.00544
-
chondroitin 4-sulfate
-
wild-type enzyme, pH 8.0, 37C
0.0651
-
chondroitin 4-sulfate
-
mutant R292A, pH 8.0, 37C
0.1
-
chondroitin 4-sulfate
-
20C
0.376
-
chondroitin 4-sulfate
-
20C
0.6
0.8
chondroitin 4-sulfate
-
recombinant enzyme, pH 8.0, 35C
0.00528
-
chondroitin 6-sulfate
-
wild-type enzyme, pH 8.0, 37C
0.0093
-
chondroitin 6-sulfate
-
-
0.0563
-
chondroitin 6-sulfate
-
mutant R292A, pH 8.0, 37C
0.121
-
chondroitin 6-sulfate
-
20C
0.215
-
chondroitin 6-sulfate
-
20C
0.6
0.8
chondroitin 6-sulfate
-
recombinant enzyme, pH 8.0, 35C
0.6
-
chondroitin 6-sulfate
-
pH 6.8, 30C, mutant enzyme Y234F
0.66
-
chondroitin 6-sulfate
-
pH 6.8, 30C, wild-type enzyme
0.38
-
chondroitin sulfate A
A9Y5J0, -
native protein
0.42
-
chondroitin sulfate A
A9Y5J0, -
recombinant protein
0.008
-
chondroitin sulfate C
-
pH 6.0, 37C
0.0104
-
chondroitin sulfate C
-
pH 6.0, 37C
0.155
-
chondroitin sulfate C
A9Y5J0, -
native protein
0.167
-
chondroitin sulfate C
A9Y5J0, -
recombinant protein
0.1755
-
hyaluronan
-
-
-
0.00119
-
hyaluronan methyl ester
-
-
0.217
-
hyaluronate
-
20C
0.239
-
hyaluronate
-
20C
0.00286
-
hyaluronic acid
-
wild-type enzyme, pH 8.0, 37C
0.0231
-
hyaluronic acid
-
mutant R292A, pH 8.0, 37C
0.234
-
hyaluronic acid
A9Y5J0, -
recombinant protein
114
-
phenyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
-
pH 6.8, 30C
114
-
phenyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
-
-
0.9
-
phenyl 4-methylumbelliferyl-beta-D-glucopyranosiduronic acid
-
pH 8.0, 30C
1
-
phenyl 4-methylumbelliferyl-beta-D-glucopyranosiduronic acid
-
pH 8.0
0.259
-
hyaluronic acid
A9Y5J0, -
native protein
additional information
-
additional information
-
effect of temperature in the Km-value
-
additional information
-
additional information
-
Km for chondroitin 4-sulfate: 0.2 g/L; Km for chondroitin 6-sulfate: 0.408 g/L, Km for hyaluronate: 0.412 g/L
-
additional information
-
additional information
-
-
-
additional information
-
additional information
-
kinetics, measurement of deuterium isotope effects
-
additional information
-
additional information
-
Km-value for chondroitin sulfate A is 0.4 mg/ml, Km-value for chondroitin sulfate C is 0.5 mg/ml, at pH 7 and 30C
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.041
-
4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiruronic acid
-
pH 6.8, 30C, wild-type enzyme
0.16
-
benzyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
-
pH 6.8
0.16
-
benzyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
-
-
0.019
-
benzyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
-
pH 6.8
0.027
-
benzyl 4-O-(2'-nitrophenyl)-beta-D-glucopyranosiduronic acid
-
pH 6.8
0.019
-
benzyl 4-O-(3',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid
-
pH 6.8
0.035
-
benzyl 4-O-(4'-chloro-2'-nitrophenyl)-beta-D-glucopyranosiduronic acid
-
pH 6.8
0.005
-
benzyl S-(2-acetamido-2-deoxy-beta-D-galactopyranosyl)-(1-4)-4-deoxy-4-thio-beta-D-glucopyranosiduronic acid
-
-
840
-
Chondroitin
-
20C
868
-
Chondroitin
-
20C
234
-
chondroitin 4-sulfate
-
recombinant enzyme, pH 8.0, 35C, dependent on expression system
308
-
chondroitin 4-sulfate
-
20C
403
-
chondroitin 4-sulfate
-
20C
345
-
chondroitin 6-sulfate
-
20C
423
480
chondroitin 6-sulfate
-
recombinant enzyme, pH 8.0, 35C, dependent on expression system
522
-
chondroitin 6-sulfate
-
20C
855
-
chondroitin 6-sulfate
-
pH 6.8, 30C, wild-type enzyme
570
-
hyaluronate
-
20C
2.3
-
phenyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
-
pH 6.8
2.3
-
phenyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid
-
-
0.0015
-
phenyl 4-methylumbelliferyl-beta-D-glucopyranosiduronic acid
-
pH 8.0
0.0016
-
phenyl 4-methylumbelliferyl-beta-D-glucopyranosiduronic acid
-
pH 8.0
1200
-
hyaluronate
-
20C
additional information
-
additional information
-
effect of temperature in the turnover number
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
pH 6.0, 37C
24
-
phenyl 4-deoxy-4,4-difluoro-beta-D-xylo-hexopyranosiduronic acid
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
3.75
-
A9Y5J0, -
-
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
-
-
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.3
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
8
-
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
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35
-
-
assay at
37
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
-
-
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.19
-
-
isoelectric focusing, pH-range 3.5-10
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Flavobacterium columnare G4
-
-
-
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
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
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
42600
-
-
gel filtration
71000
-
-
gel filtration
76000
-
-
gel filtration
78030
-
-
mass spectrometry
79500
79840
-
mass spectrometry
80000
-
A9Y5J0, -
SDS-PAGE
170000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 76000, chondroitinase I, II and III, SDS-PAGE
?
-
x * 74000, SDS-PAGE
?
-
x * 70000, recombinant enzyme, SDS-PAGE
dimer
-
2 * 84000, SDS-PAGE
dimer
-
2 * 35000, SDS-PAGE
dimer
Bacteroides stercoris HJ-15
-
2 * 84000, SDS-PAGE
-
monomer
-
1 * 75000-80000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
side-chain modification
-
carbohydrate content of chondroitinase I is 4.1%, carbohydrate content of chondroitinase II is 2.7%, carbohydrate content of chondroitinase III is 3.1%
side-chain modification
-
glycoprotein containing mannose, glucose, glucosamine and glucuronic acid in the ratio 3:5:4:2, carbohydrate content 4.1%
proteolytic modification
-
the enzyme contains a N-terminal signal peptide which is proteolytically cleaved to produce an active enzyme
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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
-
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
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
9
-
stable
6
7
-
40C, 2 h, no loss of activity
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
40
-
-
6 h, 50% loss of activity
45
-
-
pH 6.0, 15 min, stable below 45C
45
-
-
stable below
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
to homogeneity
-
using His bind column chromatography
A9Y5J0, -
recombinant enzyme
-
recombinant His-tagged active enzyme from Escherichia coli, 66fold
-
recombinant mutant from mutant Flavobacterium heparinum strain
-
wild-type enzyme, and recombinant mutant enzymes from Flavobacterium heparinum mutant strains
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli as a His-tagged fusion protein
A9Y5J0, -
the gene cslA is expressed in Escherichia coli
-
cloning and construction of mutants in Escherichia coli and expression in Flavobacterium heparinum, the Flavobacterium heparinum strains are mutated by conjugation
-
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
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
R288A
-
site-directed mutagenesis, active site residue, nearly inactive mutant
R292A
-
site-directed mutagenesis, substrate recognition residue, mutant with residual activity
R292A
-
site-directed mutagenesis, substrate recognition residue, reduced activity, action pattern is neither purely random endolytic nor purely random exolytic
Y234F
-
site-directed mutagenesis, active site residue, nearly inactive mutant
Y234F
-
mutant enzyme is inactive with chondroitin 6-sulfate
H225A
-
site-directed mutagenesis, active site residue, inactive mutant
additional information
Q59288
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 ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
useful tool for the identification and structural characterization of chondriotin and dermatan sulfates
medicine
Q59288
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