EC Number | Cloned (Comment) | Organism |
---|---|---|
4.2.2.5 | cloning and construction of mutants in Escherichia coli and expression in Flavobacterium heparinum, the Flavobacterium heparinum strains are mutated by conjugation | Pedobacter heparinus |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
4.2.2.5 | 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 | Pedobacter heparinus |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
4.2.2.5 | H225A | site-directed mutagenesis, active site residue, inactive mutant | Pedobacter heparinus |
4.2.2.5 | R288A | site-directed mutagenesis, active site residue, nearly inactive mutant | Pedobacter heparinus |
4.2.2.5 | R292A | site-directed mutagenesis, substrate recognition residue, mutant with residual activity | Pedobacter heparinus |
4.2.2.5 | Y234F | site-directed mutagenesis, active site residue, nearly inactive mutant | Pedobacter heparinus |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
4.2.2.5 | Pedobacter heparinus | Q59288 | gene cslA | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
4.2.2.5 | wild-type enzyme, and recombinant mutant enzymes from Flavobacterium heparinum mutant strains | Pedobacter heparinus |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
4.2.2.5 | [1,3-beta-D-glucuronic acid-1,4-beta-D-hexosylamine]n = n-1 4-deoxy-beta-D-gluc-4-enuronosyl-1,4-beta-D-hexosylamine + [1,3-beta-D-glucuronic acid-1,4-beta-D-hexosylamine] | 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 | Pedobacter heparinus |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
4.2.2.5 | 0.012 | - |
purified mutant R292A, substrate dermatan sulfate | Pedobacter heparinus |
4.2.2.5 | 0.028 | - |
purified mutant Y234F, substrate dermatan sulfate | Pedobacter heparinus |
4.2.2.5 | 0.031 | - |
purified mutant H225A, substrate chondroitin 4-sulfate | Pedobacter heparinus |
4.2.2.5 | 0.038 | - |
purified mutant R288A, substrate heparan sulfate | Pedobacter heparinus |
4.2.2.5 | 0.077 | - |
purified mutant R292A, substrate heparan sulfate | Pedobacter heparinus |
4.2.2.5 | 0.12 | - |
purified mutant Y234F, substrate heparan sulfate | Pedobacter heparinus |
4.2.2.5 | 0.18 | - |
purified transconjugated wild-type, substrate heparan sulfate | Pedobacter heparinus |
4.2.2.5 | 0.2 | - |
purified transconjugated wild-type, substrate dermatan sulfate | Pedobacter heparinus |
4.2.2.5 | 0.23 | - |
purified mutant Y234F, substrate chondroitin 6-sulfate | Pedobacter heparinus |
4.2.2.5 | 0.3 | - |
purified mutant Y234F, substrate hyaluronic acid | Pedobacter heparinus |
4.2.2.5 | 0.45 | - |
purified mutant R288A, substrate hyaluronic acid | Pedobacter heparinus |
4.2.2.5 | 0.53 | - |
purified mutant Y234F, substrate chondroitin 4-sulfate | Pedobacter heparinus |
4.2.2.5 | 0.55 | - |
purified mutant R288A, substrate chondroitin 6-sulfate | Pedobacter heparinus |
4.2.2.5 | 0.94 | - |
purified mutant R288A, substrate chondroitin 4-sulfate | Pedobacter heparinus |
4.2.2.5 | 3.63 | - |
purified mutant R292A, substrate hyaluronic acid | Pedobacter heparinus |
4.2.2.5 | 9.54 | - |
purified mutant R292A, substrate chondroitin 6-sulfate | Pedobacter heparinus |
4.2.2.5 | 21.2 | - |
purified mutant R292A, substrate chondroitin 4-sulfate | Pedobacter heparinus |
4.2.2.5 | 62.5 | - |
purified transconjugated wild-type, substrate chondroitin 6-sulfate | Pedobacter heparinus |
4.2.2.5 | 94.5 | - |
purified transconjugated wild-type, substrate hyaluronic acid | Pedobacter heparinus |
4.2.2.5 | 123.9 | - |
purified transconjugated wild-type, substrate chondroitin 4-sulfate | Pedobacter heparinus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
4.2.2.5 | chondroitin sulfate A | i.e. chondroitin 4-sulfate, best substrate, wild-type enzyme and mutants R292A, R288A, H225A and Y234F | Pedobacter heparinus | ? | - |
? | |
4.2.2.5 | chondroitin sulfate C | i.e. chondroitin 6-sulfate, wild-type enzyme and mutants R292A, R288A, and Y234F | Pedobacter heparinus | ? | - |
? | |
4.2.2.5 | dermatan sulfate | wild-type enzyme and mutants R292A, and Y234F | Pedobacter heparinus | ? | - |
? | |
4.2.2.5 | heparan sulfate | wild-type enzyme and mutants R292A, R288A, and Y234F | Pedobacter heparinus | ? | - |
? | |
4.2.2.5 | hyaluronic acid | wild-type enzyme and mutants R292A, R288A, and Y234F | Pedobacter heparinus | ? | - |
? | |
4.2.2.5 | additional information | Arg292 is involved in recognition of the N-acetyl and sulfate moieties of galactosamine, but does not participate directly in catalysis | Pedobacter heparinus | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
4.2.2.5 | chondroitinase AC | - |
Pedobacter heparinus |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
4.2.2.5 | 8 | - |
assay at | Pedobacter heparinus |