Information on EC 4.2.2.5 - chondroitin AC lyase

-

no beta-elimination

chondroitin sulfate degradation I (bacterial)

MetaCyc

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].

9047-57-8

-

4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiruronic acid

?

-

664604

-

?

benzyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid

?

benzyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid

?

benzyl 4-O-(2'-nitrophenyl)-beta-D-glucopyranosiduronic acid

?

-

synthetic chromogenic substrate

-

?

benzyl 4-O-(3',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid

?

-

synthetic chromogenic substrate

-

?

benzyl 4-O-(4'-chloro-2'-nitrophenyl)-beta-D-glucopyranosiduronic acid

?

-

synthetic chromogenic substrate

-

?

benzyl S-(2-acetamido-2-deoxy-beta-D-galactopyranosyl)-(1-4)-4-deoxy-4-thio-beta-D-glucopyranosiduronic acid

?

-

very slow cleavage, 9% product after 24 h

-

?

chondroitin

?

chondroitin 4-sulfate

2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose

chondroitin 4-sulfate

?

7 entries

chondroitin 4-sulfate

chondroitin 4-sulfate disaccharide

chondroitin 6-sulfate

?

10 entries

chondroitin sulfate A

?

12 entries

chondroitin sulfate A

unsaturated chondroitin A sulfate disaccharide

-

i.e. chondroitin 4-sulfate

-

?

chondroitin sulfate B

?

chondroitin sulfate C

?

14 entries

chondroitin sulfate C

unsaturated chondroitin sulfate C disaccharide

-

i.e. chondroitin 6-sulfate

product analysis

?

chondroitin sulfate D

?

chondroitin sulfate E

?

chondroitin sulfate/dermatan sulfate chain

?

-

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

701659

-

?

chondroitin/dermatan sulfate chain

?

-

chon-AC-lyase cleaves the linkage between GalNAc and D-GlcA

706673

-

?

dermatan sulfate

?

GlcNAc-D-glucuronic acid-GlcNAc-D-glucuronic acid

DELTA-4,5-uronic acid-GlcNAc + N-acetyl-D-glucosamine + DELTA-4,5-glucuronic acid

-

34095

-

34095

?

heparan sulfate

?

wild-type enzyme and mutants R292A, R288A, and Y234F

-

?

hyaluronan

?

hyaluronan methyl ester

?

-

664526

-

?

hyaluronate

?

hyaluronate

hyaluronate oligosaccharides

-

747455

-

?

hyaluronic acid

?

9 entries

methyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid

?

-

synthetic chromogenic substrate

-

?

phenyl 4-deoxy-4-fluoro-beta-D-glucopyranosiduronic acid

?

phenyl 4-methylumbelliferyl-beta-D-glucopyranosiduronic acid

?

phenyl 4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiduronic acid

?

-

-

?

proteoglycan

protein-keratan sulfate

-

34081

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

34081

?

additional information

?

-

17 entries

additional information

?

-

Ba2+

Ca2+

Co2+

-

activates more effectively than univalent cations

K+

-

activates

Mg2+

Mn2+

-

1 mM, activates

34098

Na+

-

activates

(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

Cu2+

dermatan sulfate

-

EDTA

presence of EDTA decreases the melting temperature

eicosadienoic acid

eicosanoic acid

eicosapentaenoic acid

eicosatetraenoic acid

eicosatrienoic acid

elaidic acid

Fe2+

heparin

-

lauric acid

linoleic acid

linolenic acid

methyl (9Z)-octadecenoate

myristic acid

myristoleic acid

nervonic acid

Ni2+

oleic acid

p-chloromercuriphenyl sulfonic acid

-

strong inhibition

palmitic acid

palmitoleic acid

Pb2+

-

petroselinic acid

phenyl 4-deoxy-4,4-difluoro-beta-D-xylo-hexopyranosiduronic acid

-

competitive inhibition

ricinoleic acid

Sn2+

-

stearic acid

vaccenic acid

Zn2+

additional information

-

Ca2+

20 mM, 2fold activation

chondroitin 4-sulfate

2.5fold induction of enzyme expression

649356

Na+

100 mM, 2fold activation

additional information

-

not affected by Mg2+, Mn2+, Ca2+, Co2+, and EDTA

-

6.9

4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiruronic acid

-

pH 6.8, 30°C, wild-type enzyme

664604

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

8 entries

0.38 - 0.42

chondroitin sulfate A

0.008 - 0.167

chondroitin sulfate C

0.1755

hyaluronan

-

664526

0.00119

hyaluronan methyl ester

-

664526

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

chondroitin 6-sulfate

8 entries

0.041

4-O-(2',4'-dinitrophenyl)-beta-D-glucopyranosiruronic acid

-

pH 6.8, 30°C, wild-type enzyme

664604

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

649318, 651587

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 - 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

-

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, 37°C

652640

0.0047

nervonic acid

-

pH 6.0, 37°C

652640

24

phenyl 4-deoxy-4,4-difluoro-beta-D-xylo-hexopyranosiduronic acid

-

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

111

-

chondroitin 6-sulfate

34100

123.9

purified transconjugated wild-type, substrate chondroitin 4-sulfate

21.2

purified mutant R292A, substrate chondroitin 4-sulfate

214

substrate chondroitin 6-sulfate, 39°C, pH 7.2

3.63

purified mutant R292A, substrate hyaluronic acid

3.75

A9Y5J0

-

694963

4

-

pH 7.4, 37°C

43.2

substrate hyaluronic acid, 39°C, pH 7.2

489

substrate chondroitin 4-sulfate, 39°C, pH 7.2

541

-

purified recombinant enzyme

57.03

-

purified enzyme

62.5

purified transconjugated wild-type, substrate chondroitin 6-sulfate

68

-

chondroitin 4-sulfate

34100

8.5

Arthrobacter sp. MAT3885

I6QML9

substrate chondroitin 6-sulfate, pH 7.0, 40°C

748523

9.54

purified mutant R292A, substrate chondroitin 6-sulfate

94.5

purified transconjugated wild-type, substrate hyaluronic acid

99.7

-

34098

additional information

5.7 - 6

-

6

6 - 7.5

7.2

-

7.3

-

assay at

7.5

-

8

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

35

-

assay at

37

39

-

40

45 - 50

-

50

-

20 - 25

-

activity is significantly higher at 25°C than at 20°C

681149

25 - 55

-

25°C: about 85% of maximal activity, 55°C: about 45% of maximal activity

50

-

10% residual activity

additional information

-

12

-

calculated from sequence

7.19

-

isoelectric focusing, pH-range 3.5-10

-

34098

-

Arthrobacter sp. MAT3885

-

748523

I6QML9

UniProt

Bacteroides stercoris HJ-15

human intestinal anaerobic bacterium

-

Flavobacterium columnare G4

-

34102, 663801, 681149, 729906

-

-

663801

-

Homo sapiens

-

714824

-

Paenarthrobacter aurescens 161MFSha2.1

-

-

-

-

-

747941, 748419

UniProt

-

UniProt

-

-

commercial preparation

-

physiological function

Homo sapiens

-

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

714824

6 entries

170000

-

gel filtration

35000

-

2 * 35000, SDS-PAGE

42600

-

gel filtration

34102

70000

x * 70000, recombinant enzyme, SDS-PAGE

649356

71000

-

gel filtration

74000

-

x * 74000, SDS-PAGE

34100

76000

78030

-

mass spectrometry

79500 - 79840

-

mass spectrometry

652936

80000

84000

-

2 * 84000, SDS-PAGE

?

dimer

monomer

proteolytic modification

side-chain modification

-

-

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

-

652936

-

-

34101, 34103

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 20°C, 1 day, X-ray diffraction structure determination and analysis at 2.0-2.3 A resolution

modeling of structure. Residues N153, W105, H203, Y208, Y212, R266 and E349 are involved in catalysis

I236T

H225A

site-directed mutagenesis, active site residue, inactive mutant

R288A

site-directed mutagenesis, active site residue, nearly inactive mutant

R292A

Y234F

E349A

no residual activity

H203A

no residual activity

N153A

no residual activity

R266A

no residual activity

Y212F

no residual activity

E349A

-

no residual activity

-

H203A

-

no residual activity

-

N153A

-

no residual activity

-

R266A

-

no residual activity

-

Y212F

-

no residual activity

-

additional information

5.5 - 9

-

stable

34098

6 - 7

-

40°C, 2 h, no loss of activity

34092

30

Arthrobacter sp. MAT3885

stable for 60 min

40

45

52

melting temperature in presence of 5 mM EDTA

60

protein starts unfolding at 45°C and it completely melts at 60°C

-40°C, no loss by freeze-drying in presence of 100 mM lactose or trehalose, about 60% of enzymatic activity is lost on long-term storage in the absence of both trehalose and lactose

-

-

recombinant enzyme

-

663801

recombinant His-tagged active enzyme from Escherichia coli, 66fold

-

recombinant mutant from mutant Flavobacterium heparinum strain

-

650383

to homogeneity

-

using His bind column chromatography

A9Y5J0

694963

wild-type enzyme, and recombinant mutant enzymes from Flavobacterium heparinum mutant strains

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

A9Y5J0

694963

expression in Escherichia coli

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

649356

mutant R292A, expression in Escherichia coli and conjugation to Flavobacterium heparinum to create a mutant strain

-

650383

the gene cslA is expressed in Escherichia coli

-

663801

analysis

-

useful tool for the identification and structural characterization of chondriotin and dermatan sulfates

34089

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 20°C. 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

729906

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

693771

synthesis